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Abdelrahman RS, Elnfarawy AA, Nashy AE, Abdelsalam RA, Zaghloul MS. Targeting angiogenic and proliferative mediators by montelukast & trimetazidine Ameliorates thioacetamide-induced liver fibrosis in rats. Toxicol Appl Pharmacol 2025; 495:117208. [PMID: 39716576 DOI: 10.1016/j.taap.2024.117208] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/16/2024] [Revised: 12/13/2024] [Accepted: 12/16/2024] [Indexed: 12/25/2024]
Abstract
Liver fibrosis is a significant health complication with the potential to result in serious mortality and morbidity. However, there is no standard treatment due to its complex pathogenesis. The drug montelukast reversibly and selectively antagonizes the cysteinyl-leukotrienes-1 receptor and reduces inflammation; thus, it is used in the treatment of asthma. Trimetazidine, an anti-anginal agent, selectively inhibits the activity of mitochondrial long-chain 3-ketoacyl-CoA thiolase, inhibition of free fatty acid (FFA) oxidation. This study explores the efficacy of montelukast (5 and 10 mg/kg) and trimetazidine (10-20 mg/kg) against liver fibrosis induced by thioacetamide (TAA) in rats. Impaired liver function tests were significantly improved by montelukast and trimetazidine. The antioxidant and anti-inflammatory effects of montelukast and trimetazidine were proved by the inhibition of malondialdehyde (MDA) and nitric oxide (NO) accumulation, with elevation of glutathione (GSH) and superoxide dismutase activity, decreased heat shock protein (HSP-70) expression, and a decline in interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) levels in liver tissue. Also, the antifibrotic effects were explored by reducing levels of hydroxyproline and alpha-smooth muscle actin (α-SMA) expression in liver tissue and attenuating hepatic expression of hepatic expression of angiogenic mediator vascular endothelium growth factor (VEGF) and proliferative mediator Antigen Kiel 67 (Ki-67).
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Affiliation(s)
- Rehab S Abdelrahman
- Department of Pharmacology and Toxicology, Faculty of Pharmacy, Mansoura University, 35516 Mansoura, Egypt; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Taibah University, Al-Madina Al-Munawwarah, 30001, Saudi Arabia.
| | - Ahmed A Elnfarawy
- Biotechnology Lab, Central Administration of Biological and Innovative Products and Clinical Studies, Giza, Egypt; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tanta University, Tanta, Egypt
| | | | - Ramy A Abdelsalam
- Lecturer of Pathology, Faculty of Medicine, Mansoura University, Egypt
| | - Marwa S Zaghloul
- Department of Pharmacology and Toxicology, Faculty of Pharmacy, Mansoura University, 35516 Mansoura, Egypt; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Mansoura National University, Gamasa, 7731168, Egypt
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Zhang W, Huang C, Yin T, Miao X, Deng H, Zheng R, Ren J, Chen S. Ultrasensitive US Microvessel Imaging of Hepatic Microcirculation in the Cirrhotic Rat Liver. Radiology 2022; 307:e220739. [PMID: 36413130 DOI: 10.1148/radiol.220739] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022]
Abstract
Background Liver microcirculation dysfunction plays a vital role in the occurrence and development of liver diseases, and thus, there is a clinical need for in vivo, noninvasive, and quantitative evaluation of liver microcirculation. Purpose To evaluate the feasibility of ultrasensitive US microvessel imaging (UMI) in the visualization and quantification of hepatic microvessels in healthy and cirrhotic rats. Materials and Methods In vivo studies were performed to image hepatic microvasculature by means of laparotomy in Sprague-Dawley rats (five cirrhotic and five control rats). In vivo conventional power Doppler US and ex vivo micro-CT were performed for comparison. UMI-based quantifications of perfusion, tortuosity, and integrity of microvessels were compared between the control and cirrhotic groups by using the Wilcoxon test. Spearman correlations between quantification parameters and pathologic fibrosis, perfusion function, and hepatic hypoxia were evaluated. Results UMI helped detect minute vessels below the liver capsule, as compared with conventional power Doppler US and micro-CT. With use of UMI, lower perfusion indicated by vessel density (median, 22% [IQR, 20%-28%] vs 41% [IQR, 37%-46%]; P = .008) and fractional moving blood volume (FMBV) (median, 6.4% [IQR, 4.8%-8.6%] vs 13% [IQR, 12%-14%]; P = .008) and higher tortuosity indicated by the sum of angles metric (SOAM) (median, 3.0 [IQR, 2.9-3.0] vs 2.7 [IQR, 2.6-2.9]; P = .008) were demonstrated in the cirrhotic rat group compared with the control group. Vessel density (r = 0.85, P = .003), FMBV (r = 0.86, P = .002), and median SOAM (r = -0.83, P = .003) showed strong correlations with pathologically derived vessel density labeled with dextran. Vessel density (r = -0.81, P = .005) and median SOAM (r = 0.87, P = .001) also showed strong correlations with hepatic tissue hypoxia. Conclusion Contrast-free ultrasensitive US microvessel imaging provided noninvasive in vivo imaging and quantification of hepatic microvessels in cirrhotic rat liver. © RSNA, 2022 Supplemental material is available for this article. See also the editorial by Fetzer in this issue.
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Affiliation(s)
- Wei Zhang
- From the Department of Ultrasound, Laboratory of Novel Optoacoustic (Ultrasonic) Imaging, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Rd, Guangzhou 510630, China (W.Z., T.Y., X.M., H.D., R.Z., J.R.); and Department of Radiology, Mayo Clinic College of Medicine and Science, Rochester, Minn (C.H., S.C.)
| | - Chengwu Huang
- From the Department of Ultrasound, Laboratory of Novel Optoacoustic (Ultrasonic) Imaging, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Rd, Guangzhou 510630, China (W.Z., T.Y., X.M., H.D., R.Z., J.R.); and Department of Radiology, Mayo Clinic College of Medicine and Science, Rochester, Minn (C.H., S.C.)
| | - Tinghui Yin
- From the Department of Ultrasound, Laboratory of Novel Optoacoustic (Ultrasonic) Imaging, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Rd, Guangzhou 510630, China (W.Z., T.Y., X.M., H.D., R.Z., J.R.); and Department of Radiology, Mayo Clinic College of Medicine and Science, Rochester, Minn (C.H., S.C.)
| | - Xiaoyan Miao
- From the Department of Ultrasound, Laboratory of Novel Optoacoustic (Ultrasonic) Imaging, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Rd, Guangzhou 510630, China (W.Z., T.Y., X.M., H.D., R.Z., J.R.); and Department of Radiology, Mayo Clinic College of Medicine and Science, Rochester, Minn (C.H., S.C.)
| | - Huan Deng
- From the Department of Ultrasound, Laboratory of Novel Optoacoustic (Ultrasonic) Imaging, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Rd, Guangzhou 510630, China (W.Z., T.Y., X.M., H.D., R.Z., J.R.); and Department of Radiology, Mayo Clinic College of Medicine and Science, Rochester, Minn (C.H., S.C.)
| | - Rongqin Zheng
- From the Department of Ultrasound, Laboratory of Novel Optoacoustic (Ultrasonic) Imaging, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Rd, Guangzhou 510630, China (W.Z., T.Y., X.M., H.D., R.Z., J.R.); and Department of Radiology, Mayo Clinic College of Medicine and Science, Rochester, Minn (C.H., S.C.)
| | - Jie Ren
- From the Department of Ultrasound, Laboratory of Novel Optoacoustic (Ultrasonic) Imaging, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Rd, Guangzhou 510630, China (W.Z., T.Y., X.M., H.D., R.Z., J.R.); and Department of Radiology, Mayo Clinic College of Medicine and Science, Rochester, Minn (C.H., S.C.)
| | - Shigao Chen
- From the Department of Ultrasound, Laboratory of Novel Optoacoustic (Ultrasonic) Imaging, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Rd, Guangzhou 510630, China (W.Z., T.Y., X.M., H.D., R.Z., J.R.); and Department of Radiology, Mayo Clinic College of Medicine and Science, Rochester, Minn (C.H., S.C.)
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Wang T, Lü S, Hao Y, Su Z, Long M, Cui Y. Influence of microflow on hepatic sinusoid blood flow and red blood cell deformation. Biophys J 2021; 120:4859-4873. [PMID: 34536388 DOI: 10.1016/j.bpj.2021.09.020] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/30/2021] [Revised: 06/10/2021] [Accepted: 09/10/2021] [Indexed: 01/22/2023] Open
Abstract
Hepatic sinusoids present complex anatomical structures such as the endothelial sieve pores and the Disse space, which govern the microscopic blood flow in the sinusoids and are associated with structural variations in liver fibrosis and cirrhosis. However, the contributions of the permeability of endothelial and collagen layers and the roughness of hepatocyte microvilli to the features of this microflow remain largely unknown. Here, an immersed boundary method coupled with a lattice Boltzmann method was adopted in an in vitro hepatic sinusoidal model, and flow field and erythrocyte deformation analyses were conducted by introducing three new source terms including permeability of the endothelial layer, resistance of hepatocyte microvilli and collagen layers, and deformation of red blood cells (RBCs). Numerical calculations indicated that alterations in endothelial permeability could significantly affect the flow velocity and flow rate distributions in hepatic sinusoids. Interestingly, a biphasic regulating pattern of shear stress occurred simultaneously on the surface of hepatocytes and the lower side of endothelium, i.e., the shear stress increased with increased thickness of hepatocyte microvilli and collagen layer when the endothelial permeability was high but decreased with the increase of the thickness at low endothelial permeability. Additionally, this specified microflow manipulates typical RBC deformation inside the sinusoid, yielding one-third of the variation of deformable index with varied endothelial permeability. These simulations not only are consistent with experimental measurements using in vitro liver sinusoidal chip but also elaborate the contributions of endothelial and collagen layer permeability and wall roughness. Thus, our results provide a basis for further characterizing this microflow and understanding its effects on cellular migration and deformation in the hepatic sinusoids.
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Affiliation(s)
- Tianhao Wang
- School of Mechanical Engineering, Tianjin University, Tianjin, China
| | - Shouqin Lü
- Center of Biomechanics and Bioengineering, Key Laboratory of Microgravity (National Microgravity Laboratory), Beijing Key Laboratory of Engineered Construction and Mechanobiology, Institute of Mechanics, Chinese Academy of Sciences, Beijing, China; School of Engineering Science, University of Chinese Academy of Sciences, Beijing, China
| | - Yinjing Hao
- School of Mechanical Engineering, Tianjin University, Tianjin, China
| | - Zinan Su
- School of Mechanical Engineering, Tianjin University, Tianjin, China
| | - Mian Long
- Center of Biomechanics and Bioengineering, Key Laboratory of Microgravity (National Microgravity Laboratory), Beijing Key Laboratory of Engineered Construction and Mechanobiology, Institute of Mechanics, Chinese Academy of Sciences, Beijing, China; School of Engineering Science, University of Chinese Academy of Sciences, Beijing, China.
| | - Yuhong Cui
- School of Mechanical Engineering, Tianjin University, Tianjin, China.
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Mohamed SM, Mohammed DS, Abd Elhaliem NG, Elbadry MI, Abu-Dief EE. Mangosteen Can Improve Steatohepatitis through Modulating Inflammatory and Autophagy/Apoptosis Cell Injury: An Animal Model Study. CYTOL GENET+ 2021. [DOI: 10.3103/s0095452721050091] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022]
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Cavalcante MF, Adorne MD, Turato WM, Kemmerer M, Uchiyama MK, Asbahr ACC, Alves ADCS, Farsky SHP, Drewes C, Spatti MC, Kazuma SM, Boss M, Guterres SS, Araki K, Brüne B, Namgaladze D, Pohlmann AR, Abdalla DSP. scFv-Anti-LDL(-)-Metal-Complex Multi-Wall Functionalized-Nanocapsules as a Promising Tool for the Prevention of Atherosclerosis Progression. Front Med (Lausanne) 2021; 8:652137. [PMID: 33959626 PMCID: PMC8095373 DOI: 10.3389/fmed.2021.652137] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/11/2021] [Accepted: 03/17/2021] [Indexed: 11/13/2022] Open
Abstract
Atherosclerosis can be originated from the accumulation of modified cholesterol-rich lipoproteins in the arterial wall. The electronegative LDL, LDL(-), plays an important role in the pathogenesis of atherosclerosis once this cholesterol-rich lipoprotein can be internalized by macrophages, contributing to the formation of foam cells, and provoking an immune-inflammatory response. Herein, we engineered a nanoformulation containing highly pure surface-functionalized nanocapsules using a single-chain fragment variable (scFv) reactive to LDL(-) as a ligand and assessed whether it can affect the LDL(-) uptake by primary macrophages and the progression of atherosclerotic lesions in Ldlr -/- mice. The engineered and optimized scFv-anti-LDL(-)-MCMN-Zn nanoformulation is internalized by human and murine macrophages in vitro by different endocytosis mechanisms. Moreover, macrophages exhibited lower LDL(-) uptake and reduced mRNA and protein levels of IL1B and MCP1 induced by LDL(-) when treated with this new nanoformulation. In a mouse model of atherosclerosis employing Ldlr -/- mice, intravenous administration of scFv-anti-LDL(-)-MCMN-Zn nanoformulation inhibited atherosclerosis progression without affecting vascular permeability or inducing leukocytes-endothelium interactions. Together, these findings suggest that a scFv-anti-LDL(-)-MCMN-Zn nanoformulation holds promise to be used in future preventive and therapeutic strategies for atherosclerosis.
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Affiliation(s)
- Marcela Frota Cavalcante
- Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
| | - Márcia Duarte Adorne
- Department of Organic Chemistry, Chemistry Institute, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
| | - Walter Miguel Turato
- Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
| | - Marina Kemmerer
- Faculty of Medicine, Institute of Biochemistry I, Goethe-University Frankfurt, Frankfurt, Germany
| | - Mayara Klimuk Uchiyama
- Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, São Paulo, Brazil
| | - Ana Carolina Cavazzin Asbahr
- Department of Organic Chemistry, Chemistry Institute, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
| | - Aline de Cristo Soares Alves
- Department of Production and Control of Medicines, Faculty of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
| | - Sandra Helena Poliselli Farsky
- Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
| | - Carine Drewes
- Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
| | - Marina Cecília Spatti
- Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
| | - Soraya Megumi Kazuma
- Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
| | - Marcel Boss
- Faculty of Medicine, Institute of Biochemistry I, Goethe-University Frankfurt, Frankfurt, Germany
| | - Silvia Stanisçuaski Guterres
- Department of Production and Control of Medicines, Faculty of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
| | - Koiti Araki
- Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, São Paulo, Brazil
| | - Bernhard Brüne
- Faculty of Medicine, Institute of Biochemistry I, Goethe-University Frankfurt, Frankfurt, Germany
| | - Dmitry Namgaladze
- Faculty of Medicine, Institute of Biochemistry I, Goethe-University Frankfurt, Frankfurt, Germany
| | - Adriana Raffin Pohlmann
- Department of Organic Chemistry, Chemistry Institute, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
| | - Dulcineia Saes Parra Abdalla
- Department of Clinical and Toxicological Analysis, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
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Elnfarawy AA, Nashy AE, Abozaid AM, Komber IF, Elweshahy RH, Abdelrahman RS. Vinpocetine attenuates thioacetamide-induced liver fibrosis in rats. Hum Exp Toxicol 2021; 40:355-368. [PMID: 32840391 DOI: 10.1177/0960327120947453] [Citation(s) in RCA: 38] [Impact Index Per Article: 9.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/27/2024]
Abstract
Liver fibrosis is associated with increased mortality and morbidity. However, there is not effective treatment so far. Vinpocetine (Vinpo) is a synthetic derivative of vinca alkaloid vincamine. Limited previous reports have shown some beneficial effects of Vinpo in different organ fibrosis, but the ability of Vinpo to inhibit liver fibrosis induced by thioacetamide (TAA) has not been reported, that is why we investigate the potential ability of this vinca alkaloid derivative to attenuate liver fibrosis. Hepatic fibrosis was induced in male Sprague Dawley rats by TAA (200 mg/kg; ip; 3 times/week) for 6 weeks. Daily treatments with Vinpo (10-20 mg/kg/day; orally) ameliorated TAA-induced hepatic oxidative stress and histopathological damage as indicated by a decrease in liver injury markers, LDH, hepatic MDA, and NOx levels, as well as increase anti-oxidative parameters. Besides, the anti-fibrotic efficacy of Vinpo was confirmed by decreasing hydroxyproline, and α-SMA. Also, the anti-inflammatory effect of Vinpo was explored by decreasing IL-6 and TNF-α levels. Our novel findings were that Vinpo decreased VEGF/Ki-67 expression in the liver confirming its effect on angiogenesis and proliferation. These findings reveal the anti-fibrotic effect of Vinpo against TAA-induced liver fibrosis in rats, and suggest the modulation of oxidative stress, inflammation, angiogenesis and proliferation as mechanistic cassette underlines this effect.
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Affiliation(s)
| | - Asmaa E Nashy
- 158395Faculty of Pharmacy, Mansoura University, Mansoura, Egypt
| | - Alaa M Abozaid
- 158395Faculty of Pharmacy, Mansoura University, Mansoura, Egypt
| | | | | | - Rehab S Abdelrahman
- Department of Pharmacology and Toxicology, College of Pharmacy, Taibah University, Al-Madina Al-Munawwarah, Saudi Arabia
- Department of Pharmacology and Toxicology, 158395Faculty of Pharmacy, Mansoura University, 35516, Mansoura, Egypt
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Wagner WL, Föhst S, Hock J, Kim YO, Popov Y, Schuppan D, Schladitz K, Redenbach C, Ackermann M. 3D analysis of microvasculature in murine liver fibrosis models using synchrotron radiation-based microtomography. Angiogenesis 2021; 24:57-65. [PMID: 33037487 PMCID: PMC7920893 DOI: 10.1007/s10456-020-09751-9] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/22/2020] [Accepted: 09/28/2020] [Indexed: 12/16/2022]
Abstract
Cirrhosis describes the development of excess fibrous tissue around regenerative nodules in response to chronic liver injury and usually leads to irreversible organ damage and end-stage liver disease. During the development of cirrhosis, the formation of collagenous scar tissue is paralleled by a reorganization and remodeling of the hepatic vascular system. To date, macrovascular remodeling in various cirrhosis models has been examined using three-dimensional (3D) imaging modalities, while microvascular changes have been studied mainly by two-dimensional (2D) light microscopic and electron microscopic imaging. Here, we report on the application of high-resolution 3D synchrotron radiation-based microtomography (SRμCT) for the study of the sinusoidal and capillary blood vessel system in three murine models of advanced parenchymal and biliary hepatic fibrosis. SRμCT facilitates the characterization of microvascular architecture and identifies features of intussusceptive angiogenesis in progressive liver fibrosis in a non-destructive 3D manner.
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Affiliation(s)
- Willi L Wagner
- Department of Diagnostic and Interventional Radiology, University of Heidelberg, Heidelberg, Germany
- Translational Lung Research Center, Member of the German Center for Lung Research, University of Heidelberg, Heidelberg, Germany
| | - Sonja Föhst
- Mathematics Department, Technische Universität Kaiserslautern, Kaiserslautern, Germany
| | - Jessica Hock
- Institute of Functional and Clinical Anatomy, University Medical Center of the Johannes Gutenberg-University Mainz, Johann-Joachim-Becher-Weg 13, 55128, Mainz, Germany
| | - Yong Ook Kim
- Institute of Translational Immunology and Research Center for Immune Therapy (FZI), University Medical Center of the Johannes Gutenberg-University, Mainz, Germany
| | - Yury Popov
- Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA
| | - Detlef Schuppan
- Institute of Translational Immunology and Research Center for Immune Therapy (FZI), University Medical Center of the Johannes Gutenberg-University, Mainz, Germany
- Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA
| | - Katja Schladitz
- Image Processing Department, Fraunhofer ITWM, Kaiserslautern, Germany
| | - Claudia Redenbach
- Mathematics Department, Technische Universität Kaiserslautern, Kaiserslautern, Germany
| | - Maximilian Ackermann
- Institute of Functional and Clinical Anatomy, University Medical Center of the Johannes Gutenberg-University Mainz, Johann-Joachim-Becher-Weg 13, 55128, Mainz, Germany.
- Institute of Pathology and Molecular Pathology, Helios University Clinic Wuppertal, University of Witten-Herdecke, Wuppertal, Germany.
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Lafoz E, Ruart M, Anton A, Oncins A, Hernández-Gea V. The Endothelium as a Driver of Liver Fibrosis and Regeneration. Cells 2020; 9:E929. [PMID: 32290100 PMCID: PMC7226820 DOI: 10.3390/cells9040929] [Citation(s) in RCA: 84] [Impact Index Per Article: 16.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/22/2020] [Revised: 04/05/2020] [Accepted: 04/06/2020] [Indexed: 02/07/2023] Open
Abstract
Liver fibrosis is a common feature of sustained liver injury and represents a major public health problem worldwide. Fibrosis is an active research field and discoveries in the last years have contributed to the development of new antifibrotic drugs, although none of them have been approved yet. Liver sinusoidal endothelial cells (LSEC) are highly specialized endothelial cells localized at the interface between the blood and other liver cell types. They lack a basement membrane and display open channels (fenestrae), making them exceptionally permeable. LSEC are the first cells affected by any kind of liver injury orchestrating the liver response to damage. LSEC govern the regenerative process initiation, but aberrant LSEC activation in chronic liver injury induces fibrosis. LSEC are also main players in fibrosis resolution. They maintain liver homeostasis and keep hepatic stellate cell and Kupffer cell quiescence. After sustained hepatic injury, they lose their phenotype and protective properties, promoting angiogenesis and vasoconstriction and contributing to inflammation and fibrosis. Therefore, improving LSEC phenotype is a promising strategy to prevent liver injury progression and complications. This review focuses on changes occurring in LSEC after liver injury and their consequences on fibrosis progression, liver regeneration, and resolution. Finally, a synopsis of the available strategies for LSEC-specific targeting is provided.
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Affiliation(s)
- Erica Lafoz
- Unidad de Hemodinámica Hepática, Servicio de Hepatología, Hospital Clínic, Universidad de Barcelona, Instituto de Investigaciones Biomédicas Augusto Pi Suñer (IDIBAPS), 08036 Barcelona, Spain; (E.L.); (M.R.); (A.A.); (A.O.)
| | - Maria Ruart
- Unidad de Hemodinámica Hepática, Servicio de Hepatología, Hospital Clínic, Universidad de Barcelona, Instituto de Investigaciones Biomédicas Augusto Pi Suñer (IDIBAPS), 08036 Barcelona, Spain; (E.L.); (M.R.); (A.A.); (A.O.)
| | - Aina Anton
- Unidad de Hemodinámica Hepática, Servicio de Hepatología, Hospital Clínic, Universidad de Barcelona, Instituto de Investigaciones Biomédicas Augusto Pi Suñer (IDIBAPS), 08036 Barcelona, Spain; (E.L.); (M.R.); (A.A.); (A.O.)
| | - Anna Oncins
- Unidad de Hemodinámica Hepática, Servicio de Hepatología, Hospital Clínic, Universidad de Barcelona, Instituto de Investigaciones Biomédicas Augusto Pi Suñer (IDIBAPS), 08036 Barcelona, Spain; (E.L.); (M.R.); (A.A.); (A.O.)
| | - Virginia Hernández-Gea
- Unidad de Hemodinámica Hepática, Servicio de Hepatología, Hospital Clínic, Universidad de Barcelona, Instituto de Investigaciones Biomédicas Augusto Pi Suñer (IDIBAPS), 08036 Barcelona, Spain; (E.L.); (M.R.); (A.A.); (A.O.)
- Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, 28029 Madrid, Spain
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El-Ahwany E, Mansy SS, Zoheiry M, Mourad L, Mahmoud S, Abu-Taleb H, Hassanien M, Hassan M. MicroRNA-195 vector influence on the development of gradually induced hepatocellular carcinoma in murine model. Ultrastruct Pathol 2020; 44:203-210. [PMID: 32216509 DOI: 10.1080/01913123.2020.1744783] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/24/2022]
Abstract
Background: Recent studies implicate the role of microRNAs in the pathogenesis of hepatocellular carcinoma (HCC). This study was designed to induce HCC, in an experimental model, with the prospect to study the molecular pathophysiologic changes accompanying the development of HCC and the effect of miRNA-195 vector on the process of hepatocarcinogenesis.Methodology: This study incorporated three groups of male albino mice; one control group and two other groups injected intraperitoneal with diethylnitrosamine (DEN) weekly for 12 weeks for the gradual induction of HCC. The third group was injected intra-hepatic with miR-195 vector 1 month after DEN injection. At the 8th and 12th weeks post-DEN treatment, the tumor-associated biomarkers alpha-fetoprotein (AFP), vascular endothelial growth factor (VEGF), and tumor necrosis factor-alpha (TNF-α) were assessed in the serum of all mice. Hepatic specimens were subjected to ultra-structural pathological examination as well as to caspase-3 and survivin genes expression analysis.Results: All the assessed serological and molecular parameters of HCC development, in the miRNA-195-treated group of mice, showed a significant increase, versus the DEN-treated group, whereas survivin was significantly down-regulated, in the miR-195-treated group (P < 0.001). Additionally, ultra-structural criteria of HCC were depicted, in the 12th week, in DEN-injected group, versus the 8th week, in the miRNA-195-treated group.Conclusions: Intra-hepatic injection of miRNA-195 vector induced apoptotic gene expression and suppressed anti-apoptotic gene but these favorable anti-cancer effects could not counteract the inflammatory, and subsequently, the oncogenic effect probably caused by vector administration. Therefore, further studies are required to investigate the effect of miRNA in combination with anti-inflammatory medications.
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Affiliation(s)
- Eman El-Ahwany
- Immunology Department, Theodor Bilharz Research Institute, Giza, Egypt
| | - Soheir S Mansy
- Electron Microscopy Research Department (Pathology), Theodor Bilharz Research Institute, Giza, Egypt
| | - Mona Zoheiry
- Immunology Department, Theodor Bilharz Research Institute, Giza, Egypt
| | - Lobna Mourad
- Biology Department, American University in Cairo, Cairo, Egypt
| | - Soheir Mahmoud
- Parasitology Department, Theodor Bilharz Research Institute, Giza, Egypt
| | - Hoda Abu-Taleb
- Environmental Research Department, Theodor Bilharz Research Institute, Giza, Egypt
| | - Moataz Hassanien
- Hepato-Gastroenterology Department, Theodor Bilharz Research Institute, Giza, Egypt
| | - Marwa Hassan
- Immunology Department, Theodor Bilharz Research Institute, Giza, Egypt
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Zhang Q, Jiang L, Yu J, Tian L, Guo T, Di B, Quan J, Feng J, Liu J. Palmitate up-regulates laminin expression via ROS/integrin αvβ3 pathway in HLSECs. Oncotarget 2019; 10:4083-4090. [PMID: 31289608 PMCID: PMC6609245 DOI: 10.18632/oncotarget.26937] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/14/2017] [Accepted: 03/24/2018] [Indexed: 02/05/2023] Open
Abstract
Aims/Introduction To investigate the roles of reactive oxygen species (ROS) and integrin αvβ3 in palmitate-induced laminin expression of human liver sinusoidal endothelial cells (HLSECs). Results The protein expression of integrin αv, integrin β3 and laminin are increased by palmitate in HLSECs in a time- and dose-dependent manner. NAC, the ROS inhibitor, significantly inhibited the up-regulation of protein expression of integrin αv, integrin β3 and laminin by palmitate (P < 0.05). Palmitate markedly enhanced ROS formation (P < 0.05), which was not inhibited by LM609, the antibody of integrin αvβ3. Palmitate significantly increased laminin synthesis (P < 0.05), which was attenuated by LM609 and NAC (P < 0.05). Materials and Methods HLSECs were treated with palmitate in the presence or absence of LM609 (10 μg/ml) or N-acetylcysteine (NAC) (2 mM). Expression of integrin αv, integrin β3 and laminin were measured by RT-PCR and Western blot. Immunocytochemistry were used for examining laminin expression. The generation of ROS was measured using the fluorescent signal 2',7' dichloro-fluorescein diacetate (DCFH-DA). Conclusions The results suggested that palmitate increases laminin expression through ROS/integrin αv/β3 pathway.
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Affiliation(s)
- Qi Zhang
- Department of Endocrinology, Gansu Provincial Hospital, Lanzhou City 730000, Gansu Province, China.,School of Basic Medical Sciences, Lanzhou University, Lanzhou City 730000, Gansu Province, China.,These authors contributed equally to this work
| | - Luxia Jiang
- Intensive Care Unit, Lanzhou University Second Hospital, Lanzhou City 730000, Gansu Province, China.,These authors contributed equally to this work
| | - Jing Yu
- Gansu University of Chinese Medicine, Lanzhou City 730000, Gansu Province, China
| | - Limin Tian
- Department of Endocrinology, Gansu Provincial Hospital, Lanzhou City 730000, Gansu Province, China
| | - Tiankang Guo
- Department of Endocrinology, Gansu Provincial Hospital, Lanzhou City 730000, Gansu Province, China
| | - Baoshan Di
- Emergency Department, Gansu Provincial Hospital, Lanzhou City 730000, Gansu Province, China
| | - Jinxing Quan
- Department of Endocrinology, Gansu Provincial Hospital, Lanzhou City 730000, Gansu Province, China
| | - Jing Feng
- Department of Endocrinology, Gansu Provincial Hospital, Lanzhou City 730000, Gansu Province, China
| | - Jing Liu
- Department of Endocrinology, Gansu Provincial Hospital, Lanzhou City 730000, Gansu Province, China
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11
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Portal hypertension: The desperate search for the placenta. Curr Res Transl Med 2018; 67:56-61. [PMID: 30503816 DOI: 10.1016/j.retram.2018.09.001] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/26/2018] [Revised: 09/17/2018] [Accepted: 09/30/2018] [Indexed: 12/10/2022]
Abstract
We propose that the circulatory impairments produced, in both portal hypertension and liver cirrhosis, to a certain degree resemble those characterizing prenatal life in the fetus. In fact, the left-right circulatory syndrome is common in cirrhotic patients and in the fetus. Thus, in patients with portal hypertension and chronic liver failure, the re-expression of a blood circulation comparable to fetal circulation is associated with the development of similar amniotic functions, i.e., ascites production and placenta functions, and portal vascular enteropathy. Therefore, these re-expressed embryonic functions are extra-embryonic and responsible for prenatal trophism and development.
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12
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Peeters G, Debbaut C, Friebel A, Cornillie P, De Vos WH, Favere K, Vander Elst I, Vandecasteele T, Johann T, Van Hoorebeke L, Monbaliu D, Drasdo D, Hoehme S, Laleman W, Segers P. Quantitative analysis of hepatic macro- and microvascular alterations during cirrhogenesis in the rat. J Anat 2018; 232:485-496. [PMID: 29205328 PMCID: PMC5807949 DOI: 10.1111/joa.12760] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 11/09/2017] [Indexed: 12/13/2022] Open
Abstract
Cirrhosis represents the end-stage of any persistent chronically active liver disease. It is characterized by the complete replacement of normal liver tissue by fibrosis, regenerative nodules, and complete fibrotic vascularized septa. The resulting angioarchitectural distortion contributes to an increasing intrahepatic vascular resistance, impeding liver perfusion and leading to portal hypertension. To date, knowledge on the dynamically evolving pathological changes of the hepatic vasculature during cirrhogenesis remains limited. More specifically, detailed anatomical data on the vascular adaptations during disease development is lacking. To address this need, we studied the 3D architecture of the hepatic vasculature during induction of cirrhogenesis in a rat model. Cirrhosis was chemically induced with thioacetamide (TAA). At predefined time points, the hepatic vasculature was fixed and visualized using a combination of vascular corrosion casting and deep tissue microscopy. Three-dimensional reconstruction and data-fitting enabled cirrhogenic features to extracted at multiple scales, portraying the impact of cirrhosis on the hepatic vasculature. At the macrolevel, we noticed that regenerative nodules severely compressed pliant venous vessels from 12 weeks of TAA intoxication onwards. Especially hepatic veins were highly affected by this compression, with collapsed vessel segments severely reducing perfusion capabilities. At the microlevel, we discovered zone-specific sinusoidal degeneration, with sinusoids located near the surface being more affected than those in the middle of a liver lobe. Our data shed light on and quantify the evolving angioarchitecture during cirrhogenesis. These findings may prove helpful for future targeted invasive interventions.
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Affiliation(s)
- Geert Peeters
- IBiTech – bioMMedaDepartment of Electronics and Information SystemsGhent UniversityGentBelgium
| | - Charlotte Debbaut
- IBiTech – bioMMedaDepartment of Electronics and Information SystemsGhent UniversityGentBelgium
| | - Adrian Friebel
- Interdisciplinary Centre for Bioinformatics (IZBI)University of LeipzigLeipzigGermany
- Institute of Computer ScienceUniversity of LeipzigLeipzigGermany
| | - Pieter Cornillie
- Department of MorphologyFaculty of Veterinary MedicineGhent UniversityGentBelgium
| | - Winnok H. De Vos
- Laboratory of Cell Biology and HistologyDepartment of Veterinary SciencesUniversity of AntwerpAntwerpBelgium
- Cell Systems and ImagingDepartment of Molecular BiotechnologyUniversity of GhentGentBelgium
| | - Kasper Favere
- IBiTech – bioMMedaDepartment of Electronics and Information SystemsGhent UniversityGentBelgium
| | | | - Tim Vandecasteele
- Department of MorphologyFaculty of Veterinary MedicineGhent UniversityGentBelgium
| | - Tim Johann
- Interdisciplinary Centre for Bioinformatics (IZBI)University of LeipzigLeipzigGermany
- LJLLINRIA Paris & Sorbonne Universités UPMC Univ Paris 6ParisFrance
| | - Luc Van Hoorebeke
- Centre for X‐Ray TomographyDepartment of Physics and AstronomyGhent UniversityGentBelgium
| | - Diethard Monbaliu
- Department of Microbiology and ImmunologyKU LeuvenLeuvenBelgium
- Department of Abdominal Transplant SurgeryUniversity Hospitals LeuvenLeuvenBelgium
| | - Dirk Drasdo
- Interdisciplinary Centre for Bioinformatics (IZBI)University of LeipzigLeipzigGermany
- LJLLINRIA Paris & Sorbonne Universités UPMC Univ Paris 6ParisFrance
- Leibniz Research Centre for Working Environment and Human Factors at the Technical University DortmundDortmundGermany
| | - Stefan Hoehme
- Interdisciplinary Centre for Bioinformatics (IZBI)University of LeipzigLeipzigGermany
- Institute of Computer ScienceUniversity of LeipzigLeipzigGermany
| | - Wim Laleman
- Department of Clinical and Experimental MedicineKU LeuvenLeuvenBelgium
- Department of Gastroenterology and HepatologyUniversity Hospitals LeuvenLeuvenBelgium
| | - Patrick Segers
- IBiTech – bioMMedaDepartment of Electronics and Information SystemsGhent UniversityGentBelgium
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13
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Davies T, Wythe S, O'Beirne J, Martin D, Gilbert-Kawai E. Review article: the role of the microcirculation in liver cirrhosis. Aliment Pharmacol Ther 2017; 46:825-835. [PMID: 29023881 DOI: 10.1111/apt.14279] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/25/2017] [Revised: 04/11/2017] [Accepted: 08/06/2017] [Indexed: 12/13/2022]
Abstract
BACKGROUND Intrahepatic microvascular derangements and microcirculatory dysfunction are key in the development of liver cirrhosis and its associated complications. While much has been documented relating to cirrhosis and the dysfunction of the microcirculation in the liver parenchyma, far less is known about the state of the extrahepatic microcirculation and the role this may have in the pathogenesis of multiple organ failure in end stage liver cirrhosis. AIM To provide an update on the role of the microcirculation in the pathophysiology of cirrhosis and its associated complications and briefly discuss some of the imaging techniques which may be used to directly investigate the microcirculation. METHODS A Medline literature search was conducted using the following search terms: 'cirrhosis', 'microcirculation', 'circulation', 'systemic', 'inflammation', 'peripheral', 'hepatorenal' and 'hepatopulmonary'. RESULTS Significant heterogeneous microvascular alterations exist in patients with cirrhosis. Data suggest that the systemic inflammation, associated with advanced cirrhosis, induces microcirculatory dysregulation and contributes to haemodynamic derangement. The resultant vasoconstriction and hypoperfusion in the systemic extrahepatic microvasculature, is likely to be instrumental in the pathophysiology of organ failure in decompensated cirrhosis, however the mechanistic action of vasoactive agents used to correct the circulatory disturbance of advanced cirrhosis is poorly understood. CONCLUSIONS Further research into the role of the microcirculation in patients with liver cirrhosis, will improve physicians understanding of the pathophysiology of cirrhosis, and may provide a platform for real time evaluation of an individual's microcirculatory response to vasoactive mediators, thus guiding their therapy.
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Affiliation(s)
- T Davies
- Intensive Care Department, Royal Free Hospital, London, UK.,UCLH NIHR Biomedical Research Centre, Institute of Sport and Exercise Health, University College London Centre for Altitude Space and Extreme Environment Medicine, London, UK
| | - S Wythe
- Intensive Care Department, Royal Free Hospital, London, UK.,UCLH NIHR Biomedical Research Centre, Institute of Sport and Exercise Health, University College London Centre for Altitude Space and Extreme Environment Medicine, London, UK
| | - J O'Beirne
- Department of Hepatology, Nambour General Hospital, Sunshine Coast Hospital and Health Service, Nambour, Qld, Australia
| | - D Martin
- Intensive Care Department, Royal Free Hospital, London, UK.,UCLH NIHR Biomedical Research Centre, Institute of Sport and Exercise Health, University College London Centre for Altitude Space and Extreme Environment Medicine, London, UK
| | - E Gilbert-Kawai
- Intensive Care Department, Royal Free Hospital, London, UK.,UCLH NIHR Biomedical Research Centre, Institute of Sport and Exercise Health, University College London Centre for Altitude Space and Extreme Environment Medicine, London, UK
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14
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Liebig M, Hassanzada A, Kämmerling M, Genz B, Vollmar B, Abshagen K. Microcirculatory disturbances and cellular changes during progression of hepatic steatosis to liver tumors. Exp Biol Med (Maywood) 2017; 243:1-12. [PMID: 29065724 DOI: 10.1177/1535370217738730] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
Non-alcoholic fatty liver disease is closely associated with metabolic syndrome and comprises a pathological spectrum of liver disease ranging from steatosis to steatohepatitis and can progress to fibrosis/cirrhosis and hepatocellular carcinoma. In 2013, a mouse model was described that mimics non-alcoholic fatty liver disease progression from steatohepatitis to tumors in a short time span and with high incidence. As microcirculatory disturbances play a crucial role in liver disease, the suitability of the steatosis-inflammation-tumor model for microcirculatory studies was assessed. Herein, we present a comprehensive view on morphological, microvascular, cellular, and functional aspects of non-alcoholic fatty liver disease progression in the steatosis-inflammation-tumor model using intravital microscopy, biochemical, and histological techniques. Mice develop steatohepatitis, mild fibrosis, and liver tumors at ages of 6, 12, and 20 weeks, respectively. Non-alcoholic fatty liver disease progression was accompanied by several general aspects of disease severity like increasing liver/body weight index, non-alcoholic fatty liver disease activity score, and hepatocellular apoptosis. Intravital microscopic analysis revealed significant changes in hepatic microcirculation with increasing structural alterations, elevated leukocyte adherence, and impaired nutritive perfusion. Non-alcoholic fatty liver disease was further characterized by a lower sinusoidal density with a striking rise at 20 weeks. The characteristic microcirculatory changes make the model a convenient tool for analysis of microcirculation during progression from steatosis to liver tumor. Impact statement Significant alterations of microcirculation contribute to progression of NAFLD, a chronic liver disease with increasing medical and socio-economic impact. Characterization of microcirculation in a NAFLD model reflecting all relevant stages of disease progression was still missing. Thus, we evaluated microcirculatory and cellular changes in a steatosis-inflammation-tumor model using in vivo microscopy. Analyses revealed increasing structural alterations, elevated leukocyte-endothelial interaction, and impaired nutritive perfusion. Thus, this model is suitable for further studies investigating therapeutic approaches targeting these progressive microcirculatory disturbances.
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Affiliation(s)
- Marie Liebig
- 1 Institute for Experimental Surgery, University Medicine Rostock, Rostock 18057, Germany
| | - Alireza Hassanzada
- 1 Institute for Experimental Surgery, University Medicine Rostock, Rostock 18057, Germany
| | - Malte Kämmerling
- 1 Institute for Experimental Surgery, University Medicine Rostock, Rostock 18057, Germany
| | - Berit Genz
- 1 Institute for Experimental Surgery, University Medicine Rostock, Rostock 18057, Germany.,2 QIMR Berghofer Medical Research Institute, Brisbane QLD 4006, Australia
| | - Brigitte Vollmar
- 1 Institute for Experimental Surgery, University Medicine Rostock, Rostock 18057, Germany
| | - Kerstin Abshagen
- 1 Institute for Experimental Surgery, University Medicine Rostock, Rostock 18057, Germany
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15
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Kong LJ, Li H, Du YJ, Pei FH, Hu Y, Zhao LL, Chen J. Vatalanib, a tyrosine kinase inhibitor, decreases hepatic fibrosis and sinusoidal capillarization in CCl4-induced fibrotic mice. Mol Med Rep 2017; 15:2604-2610. [PMID: 28447731 PMCID: PMC5428398 DOI: 10.3892/mmr.2017.6325] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/26/2015] [Accepted: 02/03/2017] [Indexed: 12/29/2022] Open
Abstract
Among the various consequence arising from lung injury, hepatic fibrosis is the most severe. Decreasing the effects of hepatic fibrosis remains one of the primary therapeutic challenges in hepatology. Dysfunction of hepatic sinusoidal endothelial cells is considered to be one of the initial events that occur in liver injury. Vascular endothelial growth factor signaling is involved in the progression of genotype changes. The aim of the present study was to determine the effect of the tyrosine kinase inhibitor, vatalanib, on hepatic fibrosis and hepatic sinusoidal capillarization in a carbon tetrachloride (CCl4)-induced mouse model of liver fibrosis. Liver fibrosis was induced in BALB/c mice using CCl4 by intraperitoneal injection for 6 weeks. The four experimental groups included a control, and three experimental groups involving administration of CCl4, vatalanib and a combination of the two. Histopathological staining and measuring live hydroxyproline content evaluated the extent of liver fibrosis. The expression of α-smooth muscle actin (SMA) and cluster of differentiation (CD) 34 was detected by immunohistochemistry. Collagen type I, α-SMA, transforming growth factor (TGF)-β1 and vascular endothelial growth factor receptor (VEGFR) expression levels were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The average number of fenestrae per hepatic sinusoid was determined using transmission electron microscopy. Liver fibrosis scores and hydroxyproline content were decreased in both vatalanib groups. In addition, both doses of vatalanib decreased mRNA expression levels of hepatic α-SMA, TGF-β1, collagen-1, VEGFR1, and VEGFR2. Levels of α-SMA and CD34 protein were decreased in the vatalanib group compared with the CCl4 group. There were significant differences in the number of fenestrae per sinusoid between the groups. The present study identified that administration of vatalanib was associated with decreased liver fibrosis and hepatic sinusoidal capillarization in CCl4-induced mouse models, and is a potential compound for counteracting liver fibrosis.
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Affiliation(s)
- Ling-Jian Kong
- Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
| | - Hao Li
- Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
| | - Ya-Ju Du
- Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
| | - Feng-Hua Pei
- Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
| | - Ying Hu
- Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
| | - Liao-Liao Zhao
- Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
| | - Jing Chen
- Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
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16
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Jun JH, Choi JH, Bae SH, Oh SH, Kim GJ. Decreased C-reactive protein induces abnormal vascular structure in a rat model of liver dysfunction induced by bile duct ligation. Clin Mol Hepatol 2016; 22:372-381. [PMID: 27729629 PMCID: PMC5066379 DOI: 10.3350/cmh.2016.0032] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/31/2016] [Revised: 08/05/2016] [Accepted: 08/19/2016] [Indexed: 12/23/2022] Open
Abstract
Background/Aims Chronic liver disease leads to liver fibrosis, and although the liver does have a certain regenerative capacity, this disease is associated with dysfunction of the liver vessels. C-reactive protein (CRP) is produced in the liver and circulated from there for metabolism. CRP was recently shown to inhibit angiogenesis by inducing endothelial cell dysfunction. The objective of this study was to determine the effect of CRP levels on angiogenesis in a rat model of liver dysfunction induced by bile duct ligation (BDL). Methods The diameter of the hepatic vein was analyzed in rat liver tissues using hematoxylin and eosin (H&E) staining. The expression levels of angiogenic factors, albumin, and CRP were analyzed by real-time PCR and Western blotting. A tube formation assay was performed to confirm the effect of CRP on angiogenesis in human umbilical vein endothelial cells (HUVECs) treated with lithocholic acid (LCA) and siRNA-CRP. Results The diameter of the hepatic portal vein increased significantly with the progression of cirrhosis. The expression levels of angiogenic factors were increased in the cirrhotic liver. In contrast, the expression levels of albumin and CRP were significantly lower in the liver tissue obtained from the BDL rat model than in the normal liver. The CRP level was correlated with the expression of albumin in hepatocytes treated with LCA and siRNA-CRP. Tube formation was significantly decreased in HUVECs when they were treated with LCA or a combination of LCA and siRNA-CRP. Conclusion CRP seems to be involved in the abnormal formation of vessels in hepatic disease, and so it could be a useful diagnostic marker for hepatic disease.
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Affiliation(s)
- Ji Hye Jun
- Department of Biomedical Science, CHA University, Seongnam, Korea
| | - Jong Ho Choi
- Department of Biomedical Science, CHA University, Seongnam, Korea
| | - Si Hyun Bae
- Department of Internal Medicine, Catholic University Medical College, Seoul, Korea
| | - Seh Hoon Oh
- Department of Pathology, Immunology and Laboratory Medicine, University of Florida, College of Medicine, Gainsville, FL, USA
| | - Gi Jin Kim
- Department of Biomedical Science, CHA University, Seongnam, Korea
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17
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Garbuzenko DV, Arefyev NO, Belov DV. Mechanisms of adaptation of the hepatic vasculature to the deteriorating conditions of blood circulation in liver cirrhosis. World J Hepatol 2016; 8:665-672. [PMID: 27326313 PMCID: PMC4909428 DOI: 10.4254/wjh.v8.i16.665] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/12/2016] [Revised: 04/25/2016] [Accepted: 05/27/2016] [Indexed: 02/06/2023] Open
Abstract
PubMed, EMBASE, Orphanet, MIDLINE, Google Scholar and Cochrane Library were searched for articles published between 1983 and 2015. Relevant articles were selected by using the following terms: “Liver cirrhosis”, “Endothelial dysfunction”, “Sinusoidal remodeling”, “Intrahepatic angiogenesis” and “Pathogenesis of portal hypertension”. Then the reference lists of identified articles were searched for other relevant publications as well. Besides gross hepatic structural disorders related to diffuse fibrosis and formation of regenerative nodules, the complex morphofunctional rearrangement of the hepatic microvascular bed and intrahepatic angiogenesis also play important roles in hemodynamic disturbances in liver cirrhosis. It is characterized by endothelial dysfunction and impaired paracrine interaction between activated stellate hepatocytes and sinusoidal endotheliocytes, sinusoidal remodeling and capillarization, as well as development of the collateral microcirculation. In spite of the fact that complex morphofunctional rearrangement of the hepatic microvascular bed and intrahepatic angiogenesis in liver cirrhosis are the compensatory-adaptive reaction to the deteriorating conditions of blood circulation, they contribute to progression of disease and development of serious complications, in particular, related to portal hypertension.
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18
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Liu CH, Chan KM, Chiang T, Liu JY, Chern GG, Hsu FF, Wu YH, Liu YC, Chen Y. Dual-Functional Nanoparticles Targeting CXCR4 and Delivering Antiangiogenic siRNA Ameliorate Liver Fibrosis. Mol Pharm 2016; 13:2253-62. [PMID: 27224003 DOI: 10.1021/acs.molpharmaceut.5b00913] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
The progression of liver fibrosis, an intrinsic response to chronic liver injury, is associated with hepatic hypoxia, angiogenesis, abnormal inflammation, and significant matrix deposition, leading to the development of cirrhosis and hepatocellular carcinoma (HCC). Due to the complex pathogenesis of liver fibrosis, antifibrotic drug development has faced the challenge of efficiently and specifically targeting multiple pathogenic mechanisms. Therefore, CXCR4-targeted nanoparticles (NPs) were formulated to deliver siRNAs against vascular endothelial growth factor (VEGF) into fibrotic livers to block angiogenesis during the progression of liver fibrosis. AMD3100, a CXCR4 antagonist that was incorporated into the NPs, served dual functions: it acted as a targeting moiety and suppressed the progression of fibrosis by inhibiting the proliferation and activation of hepatic stellate cells (HSCs). We demonstrated that CXCR4-targeted NPs could deliver VEGF siRNAs to fibrotic livers, decrease VEGF expression, suppress angiogenesis and normalize the distorted vessels in the fibrotic livers in the carbon tetrachloride (CCl4) induced mouse model. Moreover, blocking SDF-1α/CXCR4 by CXCR4-targeted NPs in combination with VEGF siRNA significantly prevented the progression of liver fibrosis in CCl4-treated mice. In conclusion, the multifunctional CXCR4-targeted NPs delivering VEGF siRNAs provide an effective antifibrotic therapeutic strategy.
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Affiliation(s)
- Chun-Hung Liu
- Institute of Biomedical Engineering, National Tsing Hua University , Hsinchu 30013, Taiwan, R.O.C
| | - Kun-Ming Chan
- Chang Gung Transplantation Institute, Chang Gung Memorial Hospital at Linkou, Chang Gung University College of Medicine , Taoyuan, Taiwan, R.O.C
| | - Tsaiyu Chiang
- Institute of Biomedical Engineering, National Tsing Hua University , Hsinchu 30013, Taiwan, R.O.C
| | - Jia-Yu Liu
- Institute of Biomedical Engineering, National Tsing Hua University , Hsinchu 30013, Taiwan, R.O.C
| | - Guann-Gen Chern
- Institute of Biomedical Engineering, National Tsing Hua University , Hsinchu 30013, Taiwan, R.O.C
| | - Fu-Fei Hsu
- Institute of Biomedical Sciences, Academia Sinica , Taipei, Taiwan, R.O.C
| | - Yu-Hsuan Wu
- Chang Gung Transplantation Institute, Chang Gung Memorial Hospital at Linkou, Chang Gung University College of Medicine , Taoyuan, Taiwan, R.O.C
| | - Ya-Chi Liu
- Institute of Biomedical Engineering, National Tsing Hua University , Hsinchu 30013, Taiwan, R.O.C
| | - Yunching Chen
- Institute of Biomedical Engineering, National Tsing Hua University , Hsinchu 30013, Taiwan, R.O.C
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19
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O'Brien A, China L, Massey KA, Nicolaou A, Winstanley A, Newson J, Hobbs A, Audzevich T, Gilroy DW. Bile duct-ligated mice exhibit multiple phenotypic similarities to acute decompensation patients despite histological differences. Liver Int 2016; 36:837-46. [PMID: 26012885 PMCID: PMC4869675 DOI: 10.1111/liv.12876] [Citation(s) in RCA: 22] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/05/2015] [Accepted: 05/13/2015] [Indexed: 12/16/2022]
Abstract
BACKGROUND & AIMS Patients with decompensated cirrhosis are susceptible to infection. Innate immune dysfunction and development of organ failure are considered to underlie this. A rodent model of liver disease sharing these phenotypic features would assist in vivo study of underlying mechanisms and testing of therapeutics. We evaluated three models to identify which demonstrated the greatest clinical and immunological phenotypic similarity to patients with acutely decompensated (AD) cirrhosis. METHODS We selected Bile Duct Ligation (BDL) rats at 4 weeks, BDL mice at 14 days and Carbon tetrachloride (CCl4 ) mice at 10 weeks (with studies performed 7 days after final CCl4 infection). We examined organ dysfunction, inflammatory response to carrageenan-in-paw, plasma eicosanoid concentrations, macrophage cytokine production and responses to peritoneal infection. RESULTS Bile duct ligation caused sarcopenia, liver, cardiovascular and renal dysfunction whereas CCl4 mice demonstrated no clinical abnormalities. BDL rodents exhibited depressed response to carrageenan-in-paw unlike CCl4 mice. BDL rats have slightly elevated plasma eicosanoid levels and plasma showed partial PGE2 -mediated immune suppression whereas CCl4 mice did not. Plasma NOx was elevated in patients with acute or chronic liver failure (AoCLF) compared to healthy volunteers and BDL rodents but not CCl4 mice. Elevated nitric oxide (NO) via inducible nitric oxide synthase (iNOS) mediates defective leucocyte trafficking in BDL rodent models. CONCLUSIONS We conclude that BDL mice and rats are not simply models of cholestatic liver injury but may be used to study mechanisms underlying poor outcome from infection in AD and have identified elevated NO as a potential mediator of depressed leucocyte trafficking.
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Affiliation(s)
- Alastair O'Brien
- Centre for Clinical Pharmacology and TherapeuticsDivision of MedicineUniversity College LondonLondonUK
| | - Louise China
- Centre for Clinical Pharmacology and TherapeuticsDivision of MedicineUniversity College LondonLondonUK
| | - Karen A. Massey
- Manchester Pharmacy SchoolFaculty of Medical and Human Sciencesthe University of ManchesterManchesterUK
| | - Anna Nicolaou
- Manchester Pharmacy SchoolFaculty of Medical and Human Sciencesthe University of ManchesterManchesterUK
| | - Alison Winstanley
- Department of HistopathologyUniversity College London HospitalsLondonUK
| | - Justine Newson
- Centre for Clinical Pharmacology and TherapeuticsDivision of MedicineUniversity College LondonLondonUK
| | | | - Tatsiana Audzevich
- Centre for Clinical Pharmacology and TherapeuticsDivision of MedicineUniversity College LondonLondonUK
| | - Derek W. Gilroy
- Manchester Pharmacy SchoolFaculty of Medical and Human Sciencesthe University of ManchesterManchesterUK
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20
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Aziz IA, Yacoub M, Rashid L, Solieman A. Malondialdehyde; Lipid peroxidation plasma biomarker correlated with hepatic fibrosis in human Schistosoma mansoni infection. Acta Parasitol 2015; 60:735-42. [PMID: 26408599 DOI: 10.1515/ap-2015-0105] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2014] [Accepted: 06/01/2015] [Indexed: 02/06/2023]
Abstract
Schistosomiasis is a debilitating parasitic disease, affects large number of host species. Currently affects 250-300 million people in tropic areas. Schistosoma pathogenic impact is hepatic periportal fibrosis; the parasite-induced inflammatory cellular activation promotes oxidative stress, resulting in lipid peroxidation (LPO), with subsequent increase in inflammatory mediators as malondialdehyde (MDA). This study was set up to reveal possible contribution of lipid peroxidation byproducts MDA in hepatic pathophysiology. Results displayed that MDA don't tend to change in relation with either age, nor hepatic transaminases AST & ALT, while exhibited a significant increase in MDA levels in human schistosomiasis versus control group P<0.0001 (Mn. ± St.dev. 7.77 ± 3.59, 1.21 ± 0.28 nmol/ml) respectively. Moreover; MDA plasma levels in Schistosoma infected group correlated significantly with two hepatic fibrosis parameters; (a) ultrasonography graded periportal fibrosis P< 0.0001. Levels of MDA in hepatic fibrosis grades 0, I, II, III in Schistosoma infected group were (Mn. ± St.dev. 2.8 ± 0.64, 4.3 ± 1.2, 9.3 ± 1.6 and 10.8 ± 1.3 nmol/ml) respectively, (b) serum Hyaluronic acid (HA) P<0.0001 (spearman r = 0.77) as a reliable hepatic fibrosis marker. This implies a considerable role of LPO byproducts in schistosomiasis pathogenicity, and proposing malondialdehyde as a biomarker for schistosomiasis morbidity.
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21
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Zhang Z, Li Z, Zou C, Zhang J, Zhu Y, Miao Y. Angiogenesis and proliferation of bile duct enhances ischemic tolerance in rats with cirrhosis. Int J Clin Exp Med 2015; 8:12086-12095. [PMID: 26550120 PMCID: PMC4612805] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2015] [Accepted: 08/02/2015] [Indexed: 06/05/2023]
Abstract
BACKGROUND/AIMS Primary biliary cirrhosis (PBC), an autoimmune disease of the liver, is marked by slow progressive destruction of bile ducts. These patients with PBC often undergo orthotopic liver transplantation (OLT). Ischemic bile duct lesion (IBDL) is a major source of morbidity and even mortality after OLT. Cirrhosis of the liver has a higher tolerance to ischemia than a normal liver, but the mechanism remains unknown. Angiogenesis and proliferation of bile duct often responses in bile duct ischemia, which may enhance ischemic tolerance in patients with cirrhosis. METHODOLOGY To test the hypothesis, a rat model with cirrhosis was established. Biochemical indexes of ischemic severity were measured including total bilirubin (TBIL) and direct bilirubin (DBIL). Immunohistochemical assay was performed for Ki67 (a biomarker for the proliferation of bile duct) and CD34 (a biomarker of angiogenesis). RESULTS The levels were lower for TBIL and DBIL in the bile duct from rat model with cirrhosis than that from a normal rat after ischemic surgery (P < 0.05). The levels were higher for Ki67 and CD34 from a rat model with cirrhosis than that from a normal rat after ischemic surgery (P < 0.05). CONCLUSIONS The results suggest that a liver with cirrhosis has a better ischemic tolerance than a normal liver. Angiogenesis and proliferation of bile duct enhances ischemic tolerance in rats with cirrhosis. More research on the pathogenesis of IBDLs is needed for developing more specific preventive or therapeutic strategies.
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Affiliation(s)
- Zhiqiang Zhang
- Department of General Surgery, First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China
| | - Zhennan Li
- Department of General Surgery, First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China
| | - Chen Zou
- Department of General Surgery, First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China
| | - Jingjing Zhang
- Department of General Surgery, First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China
| | - Yi Zhu
- Department of General Surgery, First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China
| | - Yi Miao
- Department of General Surgery, First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China
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22
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YOU QI, KONG LINGJIAN, LI FENGDONG, WANG HANGYU, LIU DIANGANG, PEI FENGHUA, SONG JITAO, XU JUN, CHEN JING. Human recombinant endostatin Endostar attenuates hepatic sinusoidal endothelial cell capillarization in CCl4-induced fibrosis in mice. Mol Med Rep 2015; 12:5594-600. [DOI: 10.3892/mmr.2015.4103] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/15/2014] [Accepted: 06/23/2015] [Indexed: 11/06/2022] Open
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CD147 promotes liver fibrosis progression via VEGF-A/VEGFR2 signalling-mediated cross-talk between hepatocytes and sinusoidal endothelial cells. Clin Sci (Lond) 2015. [PMID: 26201021 DOI: 10.1042/cs20140823] [Citation(s) in RCA: 40] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Although previous evidence indicates close involvement of CD147 in the pathogenesis of liver fibrosis, the underlying molecular mechanisms and its therapeutic value remain largely unknown. In the present study, we investigated the biological roles of CD147 in liver fibrosis and assessed its therapeutic value as a target molecule in the CCl4-induced liver fibrosis mouse model. We found that CD147 was highly expressed in both hepatocytes and SECs (sinusoidal endothelial cells) in fibrotic liver tissues. Additionally, it was significantly associated with the fibrosis stage. TGF-β1 (transforming growth factor β1) was found to be mainly responsible for the up-regulation of CD147. Bioinformatic and experimental data suggest a functional link between CD147 expression and VEGF-A (vascular endothelial growth factor A)/VEGR-2 (VEGF receptor 2) signalling-mediated angiogenesis in fibrotic liver tissues. Furthermore, we observed that the CD147-induced activation of the PI3K (phosphoinositide 3-kinase)/Akt signalling pathway promotes the production of VEGF-A in hepatocytes and expression of VEGFR-2 in SECs, which was found to enhance the angiogenic capability of SECs. Finally, our data indicate that blocking of CD147 using an mAb (monoclonal antibody) attenuated liver fibrosis progression via inhibition of VEGF-A/VEGFR-2 signalling and subsequent amelioration of microvascular abnormality in the CCl4-induced mouse model. Our findings suggest a novel functional mechanism that CD147 may promote liver fibrosis progression via inducing the VEGF-A/VEGFR-2 signalling pathway-mediated cross-talk between hepatocytes and SECs. New strategies based on the intervention of CD147 can be expected for prevention of liver fibrosis.
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24
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Kantari-Mimoun C, Castells M, Klose R, Meinecke AK, Lemberger UJ, Rautou PE, Pinot-Roussel H, Badoual C, Schrödter K, Österreicher CH, Fandrey J, Stockmann C. Resolution of liver fibrosis requires myeloid cell-driven sinusoidal angiogenesis. Hepatology 2015; 61:2042-55. [PMID: 25475053 DOI: 10.1002/hep.27635] [Citation(s) in RCA: 72] [Impact Index Per Article: 7.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/13/2014] [Accepted: 12/01/2014] [Indexed: 12/12/2022]
Abstract
UNLABELLED Angiogenesis is a key feature of liver fibrosis. Although sinusoidal remodeling is believed to contribute to fibrogenesis, the impact of sinusoidal angiogenesis on the resolution of liver fibrosis remains undefined. Myeloid cells, particularly macrophages, constantly infiltrate the fibrotic liver and can profoundly contribute to remodeling of liver sinusoids. We observe that the development of fibrosis is associated with decreased hepatic vascular endothelial growth factor (VEGF) expression as well as sinusoidal rarefication of the fibrotic scar. In contrast, the resolution of fibrosis is characterized by a rise in hepatic VEGF levels and revascularization of the fibrotic tissue. Genetic ablation of VEGF in myeloid cells or pharmacological inhibition of VEGF receptor 2 signaling prevents this angiogenic response and the resolution of liver fibrosis. We observe increased expression of matrix metalloproteases as well as decreased expression of tissue inhibitor of metalloproteases confined to sinusoidal endothelial cells in response to myeloid cell VEGF. Remarkably, reintroduction of myeloid cell-derived VEGF upon recovery restores collagenolytic acitivity and the resolution of fibrosis. CONCLUSION We identify myeloid cell-derived VEGF as a critical regulator of extracellular matrix degradation by liver endothelial cells, thereby unmasking an unanticipated link between angiogenesis and the resolution of fibrosis.
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Affiliation(s)
- Chahrazade Kantari-Mimoun
- Institut National de la Santé et de la Recherche Médicale (INSERM), Unit 970, Paris Cardiovascular Research Center, Paris, France
| | - Magali Castells
- Institut National de la Santé et de la Recherche Médicale (INSERM), Unit 970, Paris Cardiovascular Research Center, Paris, France
| | - Ralph Klose
- Institut National de la Santé et de la Recherche Médicale (INSERM), Unit 970, Paris Cardiovascular Research Center, Paris, France
| | - Anna-Katharina Meinecke
- Institut für Physiologie, Universitätsklinikum Essen, Universität Duisburg-Essen, Duisburg, Germany
| | - Ursula J Lemberger
- Institute of Pharmacology, Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria
| | - Pierre-Emmanuel Rautou
- Institut National de la Santé et de la Recherche Médicale (INSERM), Unit 970, Paris Cardiovascular Research Center, Paris, France.,DHU Unity, Pôle des Maladies de l'Appareil Digestif, Service d'Hépatologie, Centre de Référence des Maladies Vasculaires du Foie, Hôpital Beaujon, AP-HP, Clichy, France
| | - Hélène Pinot-Roussel
- Institut National de la Santé et de la Recherche Médicale (INSERM), Unit 970, Paris Cardiovascular Research Center, Paris, France.,Service d'Anatomie et Pathologie, Hôpital Européen Georges Pompidou, APHP, Paris, France
| | - Cécile Badoual
- Institut National de la Santé et de la Recherche Médicale (INSERM), Unit 970, Paris Cardiovascular Research Center, Paris, France.,Service d'Anatomie et Pathologie, Hôpital Européen Georges Pompidou, APHP, Paris, France
| | - Katrin Schrödter
- Institut für Physiologie, Universitätsklinikum Essen, Universität Duisburg-Essen, Duisburg, Germany
| | - Christoph H Österreicher
- Institute of Pharmacology, Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria
| | - Joachim Fandrey
- Institut für Physiologie, Universitätsklinikum Essen, Universität Duisburg-Essen, Duisburg, Germany
| | - Christian Stockmann
- Institut National de la Santé et de la Recherche Médicale (INSERM), Unit 970, Paris Cardiovascular Research Center, Paris, France
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Peeters G, Debbaut C, Cornillie P, De Schryver T, Monbaliu D, Laleman W, Segers P. A Multilevel Modeling Framework to Study Hepatic Perfusion Characteristics in Case of Liver Cirrhosis. J Biomech Eng 2015; 137:051007. [DOI: 10.1115/1.4029280] [Citation(s) in RCA: 23] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/24/2014] [Indexed: 12/16/2022]
Abstract
Liver cirrhosis represents the end-stage of different liver disorders, progressively affecting hepatic architecture, hemodynamics, and function. Morphologically, cirrhosis is characterized by diffuse fibrosis, the conversion of normal liver architecture into structurally abnormal regenerative nodules and the formation of an abundant vascular network. To date, the vascular remodeling and altered hemodynamics due to cirrhosis are still poorly understood, even though they seem to play a pivotal role in cirrhogenesis. This study aims to determine the perfusion characteristics of the cirrhotic circulation using a multilevel modeling approach including computational fluid dynamics (CFD) simulations. Vascular corrosion casting and multilevel micro-CT imaging of a single human cirrhotic liver generated detailed datasets of the hepatic circulation, including typical pathological characteristics of cirrhosis such as shunt vessels and dilated sinusoids. Image processing resulted in anatomically correct 3D reconstructions of the microvasculature up to a diameter of about 500 μm. Subsequently, two cubic samples (150 × 150 × 150 μm3) were virtually dissected from vascularized zones in between regenerative nodules and applied for CFD simulations to study the altered cirrhotic microperfusion and permeability. Additionally, a conceptual 3D model of the cirrhotic macrocirculation was developed to reveal the hemodynamic impact of regenerative nodules. Our results illustrate that the cirrhotic microcirculation is characterized by an anisotropic permeability showing the highest value in the direction parallel to the central vein (kd,zz = 1.68 × 10−13 m2 and kd,zz = 7.79 × 10−13 m2 for sample 1 and 2, respectively) and lower values in the circumferential (kd,ϑϑ = 5.78 × 10−14 m2 and kd,ϑϑ = 5.65 × 10−13 m2 for sample 1 and 2, respectively) and radial (kd,rr = 9.87 × 10−14 m2 and kd,rr = 5.13 × 10−13 m2 for sample 1 and 2, respectively) direction. Overall, the observed permeabilities are markedly higher compared to a normal liver, implying a locally decreased intrahepatic vascular resistance (IVR) probably due to local compensation mechanisms (dilated sinusoids and shunt vessels). These counteract the IVR increase caused by the presence of regenerative nodules and dynamic contraction mechanisms (e.g., stellate cells, NO-concentration, etc.). Our conceptual 3D model of the cirrhotic macrocirculation indicates that regenerative nodules severely increase the IVR beyond about 65 vol. % of regenerative nodules. Numerical modeling allows quantifying perfusion characteristics of the cirrhotic macro- and microcirculation, i.e., the effect of regenerative nodules and compensation mechanisms such as dilated sinusoids and shunt vessels. Future research will focus on the development of models to study time-dependent degenerative adaptation of the cirrhotic macro- and microcirculation.
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Affiliation(s)
- Geert Peeters
- IBiTech – bioMMeda, Department of Electronics and Information Systems, iMinds Medical IT Department, Ghent University, De Pintelaan 185 – Block B, Gent 9000, Belgium e-mail:
| | - Charlotte Debbaut
- IBiTech – bioMMeda, Department of Electronics and Information Systems, iMinds Medical IT Department, Ghent University, De Pintelaan 185 – Block B, Gent 9000, Belgium e-mail:
| | - Pieter Cornillie
- Department of Morphology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, Merelbeke 9820, Belgium
| | - Thomas De Schryver
- Centre for X-Ray Tomography, Department of Physics and Astronomy, Ghent University, Proeftuinstraat 86, Gent 9000, Belgium
| | - Diethard Monbaliu
- Department of Microbiology and Immunology, Abdominal Transplant Surgery, University Hospitals Leuven, KU Leuven, Herestraat 49, Leuven 3000, Belgium
| | - Wim Laleman
- Department of Liver and Biliopancreatic Disorders, Hepatology, University Hospitals Leuven, KU Leuven, Herestraat 49, Leuven 3000, Belgium
| | - Patrick Segers
- IBiTech – bioMMeda, Department of Electronics and Information Systems, iMinds Medical IT Department, Ghent University, De Pintelaan 185 – Block B, Gent 9000, Belgium e-mail:
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26
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Heymann F, Niemietz PM, Peusquens J, Ergen C, Kohlhepp M, Mossanen JC, Schneider C, Vogt M, Tolba RH, Trautwein C, Martin C, Tacke F. Long term intravital multiphoton microscopy imaging of immune cells in healthy and diseased liver using CXCR6.Gfp reporter mice. J Vis Exp 2015:52607. [PMID: 25866988 PMCID: PMC4401387 DOI: 10.3791/52607] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022] Open
Abstract
Liver inflammation as a response to injury is a highly dynamic process involving the infiltration of distinct subtypes of leukocytes including monocytes, neutrophils, T cell subsets, B cells, natural killer (NK) and NKT cells. Intravital microscopy of the liver for monitoring immune cell migration is particularly challenging due to the high requirements regarding sample preparation and fixation, optical resolution and long-term animal survival. Yet, the dynamics of inflammatory processes as well as cellular interaction studies could provide critical information to better understand the initiation, progression and regression of inflammatory liver disease. Therefore, a highly sensitive and reliable method was established to study migration and cell-cell-interactions of different immune cells in mouse liver over long periods (about 6 hr) by intravital two-photon laser scanning microscopy (TPLSM) in combination with intensive care monitoring. The method provided includes a gentle preparation and stable fixation of the liver with minimal perturbation of the organ; long term intravital imaging using multicolor multiphoton microscopy with virtually no photobleaching or phototoxic effects over a time period of up to 6 hr, allowing tracking of specific leukocyte subsets; and stable imaging conditions due to extensive monitoring of mouse vital parameters and stabilization of circulation, temperature and gas exchange. To investigate lymphocyte migration upon liver inflammation CXCR6.gfp knock-in mice were subjected to intravital liver imaging under baseline conditions and after acute and chronic liver damage induced by intraperitoneal injection(s) of carbon tetrachloride (CCl4). CXCR6 is a chemokine receptor expressed on lymphocytes, mainly on Natural Killer T (NKT)-, Natural Killer (NK)- and subsets of T lymphocytes such as CD4 T cells but also mucosal associated invariant (MAIT) T cells1. Following the migratory pattern and positioning of CXCR6.gfp+ immune cells allowed a detailed insight into their altered behavior upon liver injury and therefore their potential involvement in disease progression.
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Affiliation(s)
- Felix Heymann
- Department of Medicine III, RWTH University-Hospital Aachen;
| | | | | | - Can Ergen
- Department of Medicine III, RWTH University-Hospital Aachen
| | | | | | | | - Michael Vogt
- IZKF Aachen Core Facility "Two-Photon Imaging", RWTH University-Hospital Aachen
| | - Rene H Tolba
- Institute for Laboratory Animal Science & Experimental Surgery, RWTH Aachen University
| | | | | | - Frank Tacke
- Department of Medicine III, RWTH University-Hospital Aachen;
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27
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Debbaut C, Segers P, Cornillie P, Casteleyn C, Dierick M, Laleman W, Monbaliu D. Analyzing the human liver vascular architecture by combining vascular corrosion casting and micro-CT scanning: a feasibility study. J Anat 2014; 224:509-17. [PMID: 24433401 DOI: 10.1111/joa.12156] [Citation(s) in RCA: 63] [Impact Index Per Article: 5.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 12/07/2013] [Indexed: 11/26/2022] Open
Abstract
Although a full understanding of the hepatic circulation is one of the keys to successfully perform liver surgery and to elucidate liver pathology, relatively little is known about the functional organization of the liver vasculature. Therefore, we materialized and visualized the human hepatic vasculature at different scales, and performed a morphological analysis by combining vascular corrosion casting with novel micro-computer tomography (CT) and image analysis techniques. A human liver vascular corrosion cast was obtained by simultaneous resin injection in the hepatic artery (HA) and portal vein (PV). A high resolution (110 μm) micro-CT scan of the total cast allowed gathering detailed macrovascular data. Subsequently, a mesocirculation sample (starting at generation 5; 88 × 68 × 80 mm³) and a microcirculation sample (terminal vessels including sinusoids; 2.0 × 1.5 × 1.7 mm³) were dissected and imaged at a 71-μm and 2.6-μm resolution, respectively. Segmentations and 3D reconstructions allowed quantifying the macro- and mesoscale branching topology, and geometrical features of HA, PV and hepatic venous trees up to 13 generations (radii ranging from 13.2 mm to 80 μm; lengths from 74.4 mm to 0.74 mm), as well as microvascular characteristics (mean sinusoidal radius of 6.63 μm). Combining corrosion casting and micro-CT imaging allows quantifying the branching topology and geometrical features of hepatic trees using a multiscale approach from the macro- down to the microcirculation. This may lead to novel insights into liver circulation, such as internal blood flow distributions and anatomical consequences of pathologies (e.g. cirrhosis).
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Affiliation(s)
- Charlotte Debbaut
- Biofluid, Tissue and Solid Mechanics for Medical Applications, Institute Biomedical Technology, Department of Electronics and Information Systems, iMinds Future Health Department, Ghent University, Gent, Belgium
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28
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Yan J, Kang Y, Xu S, Ong LLS, Zhuo S, Bunte RM, Chen N, Asada HH, So PTC, Wanless IR, Yu H. In vivo label-free quantification of liver microcirculation using dual-modality microscopy. JOURNAL OF BIOMEDICAL OPTICS 2014; 19:116006. [PMID: 25388810 DOI: 10.1117/1.jbo.19.11.116006] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/18/2014] [Accepted: 10/21/2014] [Indexed: 05/26/2023]
Abstract
Microcirculation lesion is a common symptom of chronic liver diseases in the form of vasculature deformation and circulation alteration. In acute to chronic liver diseases such as biliary atresia, microcirculation lesion can have an early onset. Detection of microcirculation lesion is meaningful for studying the progression of liver disease. We have combined wide-field fluorescence microscopy and a laser speckle contrast technique to characterize hepatic microcirculation in vivo without labeling in a bile-duct ligation rat fibrosis model of biliary atresia. Through quantitative image analysis of four microcirculation parameters, we observed significant microcirculation lesion in the early to middle stages of fibrosis. This bimodal imaging method is useful to assess hepatic microcirculation lesion for the study of liver diseases.
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Affiliation(s)
- Jie Yan
- BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore, SingaporebInstitute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore, Singapor
| | - Yuzhan Kang
- BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore, SingaporedZhujiang Hospital of Southern Medical University, Department of Critical Care Medicine, 253 Gongye
| | - Shuoyu Xu
- BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore, SingaporebInstitute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore, Singapor
| | - Lee-Ling S Ong
- BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore, Singapore
| | - Shuangmu Zhuo
- BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore, SingaporeeFujian Normal University, Institute of Laser and Optoelectronics Technology, Fuzhou 350007, China
| | - Ralph M Bunte
- Duke-NUS Graduate Medical School Singapore, 8 College Road, 169857 Singapore, Singapore
| | - Nanguang Chen
- National University of Singapore, Faculty of Engineering, Department of bioengineering, 9 Engineering Drive 1,117574 Singapore, Singapore
| | - H Harry Asada
- BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore, SingaporehMassachusetts Institute of Technology, Department of Mechanical Engineering, 77 Massachusetts Avenu
| | - Peter T C So
- BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore, SingaporehMassachusetts Institute of Technology, Department of Mechanical Engineering, 77 Massachusetts Avenu
| | - Ian R Wanless
- Dalhousie University, Victoria General Hospital, Department of Pathology, Halifax, Nova Scotia, B3H 2Y9 Canada
| | - Hanry Yu
- BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #04-13/14 Enterprise Wing, 138602 Singapore, SingaporebInstitute of Bioengineering and Nanotechnology, 31 Biopolis Way, #04-01, 138669 Singapore, Singapor
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Abstract
Angiogenesis, defined as the formation of new microvasculature from preexisting blood vessels and mature endothelial cells, plays a major role in wound healing and scar formation, and it is associated with inflammatory responses. Angiogenesis can occur in physiological conditions, such as during liver regeneration, and in pathological situations, such as during the progression of fibrosis to cirrhosis and also during tumor angiogenesis. Cellular cross-talk among liver sinusoidal endothelial cells (LSECs), hepatic stellate cells and hepatocytes is believed to play an important role in the angiogenesis process during both liver regeneration and development of cirrhosis. In addition to mature endothelial cells, bone marrow (BM)-derived circulating endothelial progenitor cells (EPCs) have been recently identified for their contribution to post-natal vasculogenesis/angiogenesis. In vivo, EPCs are mobilized into the peripheral blood in response to tissue ischemia or traumatic injury, migrate to the sites of injured endothelium and differentiate into mature endothelial cells. In our recent studies, we have explored the role of EPC-mediated angiogenesis in liver regeneration and/or cirrhosis. Results have demonstrated significantly increased endogenous levels of circulating EPCs in cirrhotic patients in comparison to the controls. Also, EPCs from cirrhotic patients have been observed to stimulate substantial angiogenesis by resident LSECs in vitro via paracrine factors such as vascular endothelial growth factor and platelet-derived growth factor. This review gives an overview of the angiogenesis process in liver regeneration and disease and discusses a new mechanism for intrahepatic angiogenesis mediated by BM-derived EPCs.
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Affiliation(s)
- Savneet Kaur
- School of Biotechnology, Gautam Buddha University, Greater Noida, 201312, UP, India.
| | - K Anita
- School of Biotechnology, Gautam Buddha University, Greater Noida, 201312, UP, India
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30
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Li YL, Song ZJ, Fang H, Peng W, Gao JM, Shen T. Influence of treatment with loureirin on rat hepatic stellate cells in vitro. Shijie Huaren Xiaohua Zazhi 2013; 21:87-92. [DOI: 10.11569/wcjd.v21.i1.87] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To investigate the effects of treatment with loureirin A/B on rat hepatic stellate cells (HSCs) in vitro.
METHODS: HSC-T6 cells were cultured in 96-well plates for 24 h and then incubated with different concentrations of loureirin A/B for 48 h. MTT assay was used to assess the proliferation of HSCs. The contents of hyaluronic acid, laminin and collagen type IV in culture supernatants were measured by radioimmunoassay. The mRNA expression of blood vessel endothelium (VEGF165) and histoplasma tissue inhibitory factor (HIF-1) was detected by real-time fluorescence quantitative PCR.
RESULTS: The half maximal inhibitory concentrations (IC50) of loureirin A/B were 0.3 and 0.1 µg/µL respectively. Treatment with loureirin A/B significantly inhibited HSC proliferation, the secretion of hyaluronic acid, laminin and collagen type IV (31.1 ± 4.32 vs 43.05 ± 4.96, 441.28 ± 25.38 vs 302.98 ± 29.59, 17.96 ± 3.00 vs 25.23 ± 4.96, all P < 0.05), and the mRNA expression of VEGF165 and HIF-1. The effects of loureirin B were stronger than those of loureirin A.
CONCLUSION: Treatment with loureirin A/B could significantly inhibit the proliferation of HSCs, the secretion of hyaluronic acid, laminin and collagen type IV, and the mRNA expression of VEGF165 mRNA and HIF-1 mRNA, and the effects of loureirin B were stronger than those of loureirin A.
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31
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Debbaut C, Vierendeels J, Siggers JH, Repetto R, Monbaliu D, Segers P. A 3D porous media liver lobule model: the importance of vascular septa and anisotropic permeability for homogeneous perfusion. Comput Methods Biomech Biomed Engin 2012; 17:1295-310. [PMID: 23237543 DOI: 10.1080/10255842.2012.744399] [Citation(s) in RCA: 32] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
The hepatic blood circulation is complex, particularly at the microcirculatory level. Previously, 2D liver lobule models using porous media and a 3D model using real sinusoidal geometries have been developed. We extended these models to investigate the role of vascular septa (VS) and anisotropic permeability. The lobule was modelled as a hexagonal prism (with or without VS) and the tissue was treated as a porous medium (isotropic or anisotropic permeability). Models were solved using computational fluid dynamics. VS inclusion resulted in more spatially homogeneous perfusion. Anisotropic permeability resulted in a larger axial velocity component than isotropic permeability. A parameter study revealed that results are most sensitive to the lobule size and radial pressure drop. Our model provides insight into hepatic microhaemodynamics, and suggests that inclusion of VS in the model leads to perfusion patterns that are likely to reflect physiological reality. The model has potential for applications to unphysiological and pathological conditions.
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Affiliation(s)
- Charlotte Debbaut
- a Biofluid, Tissue and Solid Mechanics for Medical Applications (bioMMeda), Institute Biomedical Technology, Ghent University , Campus Heymans - Blok B, De Pintelaan 185, 9000 Gent , Belgium
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32
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Yu LL, Zhao JM, Jiang WW. Portal vein infusion of recombinant vascular endothelial growth factor 165 mitigates liver fibrosis in cirrhotic rats. Shijie Huaren Xiaohua Zazhi 2011; 19:2214-2219. [DOI: 10.11569/wcjd.v19.i21.2214] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To evaluate the effect of portal vein infusion of recombinant vascular endothelial growth factor (VEGF) 165 on liver fibrosis in rats with cirrhosis.
METHODS: Fifty male SD rats were randomly divided into normal group (n = 10) and model group (n = 40). The model group was used to induce cirrhosis using the thioacetamide approach. After 10 wk, 25 cirrhotic rats were randomly divided into experimental group (n = 15) and model control group (n = 10). The experimental group was intubated for implantation of an Alzet osmotic pump, which was used to infuse recombinant VEGF165 via the portal vein for 2 wk. The normal group and model control group underwent sham operation. All rats were killed after 2 wk, and HE staining was used to observe the pathological changes in liver tissue. Serum hyaluronic acid and laminin were measured using radioimmunoassay method. Immunohistochemistry was used to detect the expression of type I and type IV collagen in the liver.
RESULTS: Degeneration and necrosis of liver cells, diffuse proliferation of fibrous connective tissue and formation of pseudo lobules occurred in the model control group. In the experimental group, degeneration and necrosis of liver cells were milder and the rate of liver fibrosis was improved significantly compared to the model control group (P < 0.01). Compared to the normal group, serum hyaluronic acid and laminin concentrations increased significantly in the model control group (P < 0.01). However, serum concentrations of hyaluronic acid and laminin was significantly lower in the experimental group than in the model control group (412.63 μg/L ± 85.18 μg/L vs 741.60 μg/L ± 72.83 μg/L; 58.87 μg/L ± 5.46 μg/L vs 92.80 μg/L ± 8.41 μg/L; both P < 0.01). The expression levels of type I and type IV collagen in the liver was significantly lower in the experimental group than in the model control group (6.84 ± 0.96, 8.25 ± 0.82 vs 18.38 ± 1.86, 20.86 ± 2.48, all P < 0.01).
CONCLUSION: Portal vein infusion of VEGF165 can relieve liver fibrosis in rats with cirrhosis.
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Cohen JI, Nagy LE. Pathogenesis of alcoholic liver disease: interactions between parenchymal and non-parenchymal cells. J Dig Dis 2011; 12:3-9. [PMID: 21091930 PMCID: PMC5061145 DOI: 10.1111/j.1751-2980.2010.00468.x] [Citation(s) in RCA: 58] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
The development of alcoholic liver disease (ALD) is a complex process involving both the parenchymal and non-parenchymal cells in the liver. The impact of ethanol on hepatocytes can be characterized as a condition of organelle stress with multifactorial changes in hepatocellular function accumulating during ethanol exposure. These changes include oxidative stress, mitochondrial dysfunction, decreased methylation capacity, endoplasmic reticulum stress, impaired vesicular trafficking and altered proteasome function. Injury to hepatocytes is attributed, in part, to ethanol metabolism by the hepatocytes. Changes in the structural integrity of hepatic sinusoidal endothelial cells, as well as enhanced inflammation in the liver during ethanol exposure are also important contributors to injury. Activation of hepatic stellate cells initiates the deposition of extracellular matrix proteins characteristic of fibrosis. Kupffer cells, the resident macrophages in the liver, are particularly critical to the onset of ethanol-induced liver injury. Chronic ethanol exposure sensitizes Kupffer cells to activation by lipopolysaccharides via toll-like receptor 4. This sensitization enhances the production of inflammatory mediators, such as tumor necrosis factor-α and reactive oxygen species that contribute to hepatocyte dysfunction, necrosis and apoptosis of hepatocytes and the generation of extracellular matrix proteins leading to fibrosis. In this review we provide an overview of the complex interactions between parenchymal and non-parenchymal cells in the liver during the progression of ethanol-induced liver injury.
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Affiliation(s)
- Jessica I. Cohen
- Department of Pathobiology, Cleveland Clinic, Cleveland OH 44195,Department of Nutrition, Case Western Reserve University, Cleveland OH 44120
| | - Laura E. Nagy
- Department of Pathobiology, Cleveland Clinic, Cleveland OH 44195,Department of Gastroenterology and Hepatology, Cleveland Clinic, Cleveland OH 44195,Department of Nutrition, Case Western Reserve University, Cleveland OH 44120
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Huebert RC, Jagavelu K, Liebl AF, Huang BQ, Splinter PL, LaRusso NF, Urrutia RA, Shah VH. Immortalized liver endothelial cells: a cell culture model for studies of motility and angiogenesis. J Transl Med 2010; 90:1770-81. [PMID: 20644520 PMCID: PMC2992582 DOI: 10.1038/labinvest.2010.132] [Citation(s) in RCA: 46] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022] Open
Abstract
Hepatic sinusoidal endothelial cells (HSECs) are a unique subpopulation of fenestrated endothelial cells lining the hepatic sinusoids and comprising the majority of endothelial cells within the liver. HSECs not only have important roles in blood clearance, vascular tone, and immunity, but also undergo pathological changes, contributing to fibrosis, angiogenesis, and portal hypertension. There are few cell culture models for in vitro studies of motility and angiogenesis as primary cells are time-consuming to isolate, are limited in number, and often lack features of pathological vasculature. The aim of this study was to generate an immortalized cell line derived from HSECs that mimic pathological vasculature and allows detailed molecular interventions to be pursued. HSECs were isolated from mouse liver using CD31-based immunomagnetic separation, immortalized with SV40 large T-antigen, and subcloned on the basis of their ability to endocytose the acetylated low-density lipoprotein (AcLDL). The resulting cell line, transformed sinusoidal endothelial cells (TSECs), maintains an endothelial phenotype as well as some HSEC-specific features. This is evidenced by typical microscopic features of endothelia, including formation of lamellipodia and filopodia, and a cobblestone morphology of cell monolayers. Electron microscopy showed maintenance of a limited number of fenestrae organized in sieve plates. TSECs express numerous endothelia-specific markers, including CD31 and von Willebrand's factor (vWF), as detected by PCR array, immunoblotting, and immunofluorescence (IF). Functionally, TSECs maintain a number of key endothelial features, including migration in response to angiogenic factors, formation of vascular tubes, endocytosis of AcLDL, and remodeling of extracellular matrix. Their phenotype most closely resembles the pathological neovasculature associated with chronic liver disease, in which cells become proliferative, defenestrated, and angiogenic. Importantly, the cells can be transduced efficiently with viral vectors. TSECs should provide a reproducible cell culture model for high-throughput in vitro studies pertaining to a broad range of liver endothelial cell functions, but likely broader endothelial cell biology as well.
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Affiliation(s)
- Robert C. Huebert
- Gastroenterology Research Unit, Mayo Clinic and Foundation, Rochester, MN 55905,Department of Internal Medicine, Mayo Clinic and Foundation, Rochester, MN 55905
| | - Kumaravelu Jagavelu
- Gastroenterology Research Unit, Mayo Clinic and Foundation, Rochester, MN 55905
| | - Ann F. Liebl
- Gastroenterology Research Unit, Mayo Clinic and Foundation, Rochester, MN 55905
| | - Bing Q. Huang
- Center for Basic Research in Digestive Diseases, Mayo Clinic and Foundation, Rochester, MN 55905
| | - Patrick L. Splinter
- Center for Basic Research in Digestive Diseases, Mayo Clinic and Foundation, Rochester, MN 55905
| | - Nicholas F. LaRusso
- Center for Basic Research in Digestive Diseases, Mayo Clinic and Foundation, Rochester, MN 55905,Department of Internal Medicine, Mayo Clinic and Foundation, Rochester, MN 55905,Mayo Clinic Center for Cell Signaling, Mayo Clinic and Foundation, Rochester, MN 55905
| | - Raul A. Urrutia
- Gastroenterology Research Unit, Mayo Clinic and Foundation, Rochester, MN 55905,Department of Internal Medicine, Mayo Clinic and Foundation, Rochester, MN 55905
| | - Vijay H. Shah
- Gastroenterology Research Unit, Mayo Clinic and Foundation, Rochester, MN 55905,Department of Internal Medicine, Mayo Clinic and Foundation, Rochester, MN 55905,Mayo Clinic Center for Cell Signaling, Mayo Clinic and Foundation, Rochester, MN 55905
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Heindryckx F, Mertens K, Charette N, Vandeghinste B, Casteleyn C, Van Steenkiste C, Slaets D, Libbrecht L, Staelens S, Starkel P, Geerts A, Colle I, Van Vlierberghe H. Kinetics of angiogenic changes in a new mouse model for hepatocellular carcinoma. Mol Cancer 2010; 9:219. [PMID: 20727157 PMCID: PMC2936339 DOI: 10.1186/1476-4598-9-219] [Citation(s) in RCA: 34] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/04/2009] [Accepted: 08/20/2010] [Indexed: 02/07/2023] Open
Abstract
BACKGROUND The increasing incidence of hepatocellular carcinoma in Western countries has led to an expanding interest of scientific research in this field. Therefore, a vast need of experimental models that mimic the natural pathogenesis of hepatocellular carcinoma (HCC) in a short time period is present. The goal of our study was (1) to develop an efficient mouse model for HCC research, in which tumours develop in a natural background of fibrosis and (2) to assess the time-dependent angiogenic changes in the pathogenesis of HCC. METHODS Weekly intraperitoneal injections with the hepatocarcinogenic compound N-nitrosodiethylamine was applied as induction method and samples were taken at several time points to assess the angiogenic changes during the progression of HCC. RESULTS The N-nitrosodiethylamine-induced mouse model provides well vascularised orthotopic tumours after 25 weeks. It is a representative model for human HCC and can serve as an excellent platform for the development of new therapeutic targets.
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Affiliation(s)
- Femke Heindryckx
- Department of Gastroenterology and Hepatology, Ghent University Hospital, Ghent, Belgium
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Abstract
PURPOSE OF REVIEW Patients with cirrhosis have total extracellular fluid overload but central effective circulating hypovolaemia. The resulting neurohumoral compensatory response favours the accumulation of fluids into the peritoneal cavity (ascites) and may hinder renal perfusion (hepatorenal syndrome). Their deranged systemic haemodynamics (hyperdynamic circulatory syndrome) is characterized by elevated cardiac output with decreased systemic vascular resistance and low blood pressure. RECENT FINDINGS Molecular and biological mechanisms determining cirrhosis-induced haemodynamic alterations are progressively being elucidated. The need for a goal-directed assessment of volume resuscitation (especially with volumetric techniques) in patients with cirrhosis is becoming more and more evident. The role of fluid expansion with albumin and the use of splanchnic vasopressors in a variety of cirrhosis-related conditions has recently been investigated. SUMMARY The response to fluid loading in patients with advanced cirrhosis is abnormal, primarily resulting in expansion of their noncentral blood volume compartment. Colloid solutions, in particular albumin, are best used in these patients. Albumin may be effective in preventing the haemodynamic derangements associated with large-volume paracentesis (paracentesis-induced circulatory dysfunction), in preventing renal failure during spontaneous bacterial peritonitis and, in association with splanchnic vasopressors, in caring for patients with the hepatorenal syndrome.
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