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1
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O'Flaherty C. Redox signaling regulation in human spermatozoa: a primary role of peroxiredoxins. Asian J Androl 2025:00129336-990000000-00281. [PMID: 39902615 DOI: 10.4103/aja2024126] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/05/2024] [Accepted: 12/24/2024] [Indexed: 02/05/2025] Open
Abstract
ABSTRACT Reactive oxygen species (ROS) play a dual role in mammalian spermatozoa. At high levels, they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein, lipids, and sperm DNA. This oxidative damage is associated with male infertility. On the other hand, when ROS are produced at low levels, they participate in the redox signaling necessary for sperm capacitation. Capacitation-associated ROS are produced by the sperm oxidase, whose identity is still elusive, located in the plasma membrane of the spermatozoon. ROS, such as superoxide anion, hydrogen peroxide, nitric oxide, and peroxynitrite, activate protein kinases and inactivate protein phosphatases with the net increase of specific phosphorylation events. Peroxiredoxins (PRDXs), antioxidant enzymes that fight against oxidative stress, regulate redox signaling during capacitation. Among them, PRDX6, which possesses peroxidase and calcium-independent phospholipase A2 (iPLA2) activities, is the primary regulator of redox signaling and the antioxidant response in human spermatozoa. The lysophosphatidic acid signaling is essential to maintain sperm viability by activating the phosphatidylinositol 3-kinase/protein kinase (PI3K/AKT) pathway, and it is regulated by PRDX6 iPLA2, protein kinase C (PKC), and receptor-type protein tyrosine kinase. The understanding of redox signaling is crucial to pave the way for novel diagnostic tools and treatments of male infertility.
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Affiliation(s)
- Cristian O'Flaherty
- Department of Surgery (Urology Division), Faculty of Medicine and Health Sciences, McGill University, Montréal, QC H4A 3J1, Canada
- Department of Pharmacology and Therapeutics, Faculty of Medicine and Health Sciences, McGill University, Montréal, QC H3G 1Y6, Canada
- The Research Institute, McGill University Health Centre, Montréal, QC H4A 3J1, Canada
- Department of Anatomy and Cell Biology, Faculty of Medicine and Health Sciences, McGill University, Montréal, QC H3A 0C7, Canada
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2
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Zhang F, Chen F, Shen R, Chen YX, Zhao Z, Zhang B, Fang J. Naphthalimide Fluorescent Skeleton for Facile and Accurate Quantification of Glutathione. Anal Chem 2023; 95:4301-4309. [PMID: 36812128 DOI: 10.1021/acs.analchem.2c04098] [Citation(s) in RCA: 16] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/24/2023]
Abstract
Glutathione (GSH), the most prevalent nonprotein thiol in biological systems, acts as both an antioxidant to manipulate intracellular redox homeostasis and a nucleophile to detoxify xenobiotics. The fluctuation of GSH is closely related to the pathogenesis of diverse diseases. This work reports the construction of a nucleophilic aromatic substitution-type probe library based on the naphthalimide skeleton. After an initial evaluation, the compound R13 was identified as a highly efficient GSH fluorescent probe. Further studies demonstrate that R13 could readily quantify GSH in cells and tissues via a straightforward fluorometric assay with a comparable accuracy to the results from the HPLC. We then used R13 to quantify the content of GSH in mouse livers after X-ray irradiation, revealing that irradiation-induced oxidative stress leads to the increase of oxidized GSH (GSSG) and depletion of GSH. In addition, probe R13 was also applied to investigate the alteration of the GSH level in the Parkinson's mouse brains, showing a decrease of GSH and an increase of GSSG in Parkinson's mouse brains. The convenience of the probe in quantifying GSH in biological samples facilitates further understanding of the fluctuation of the GSH/GSSG ratio in diseases.
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Affiliation(s)
- Fang Zhang
- State Key Laboratory of Applied Organic Chemistry and College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China
| | - Fan Chen
- State Key Laboratory of Applied Organic Chemistry and College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China
| | - Ruipeng Shen
- State Key Laboratory of Applied Organic Chemistry and College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China
| | - Ya-Xiong Chen
- Key Laboratory of Space Radiobiology of Gansu Province & CAS Key Laboratory of Heavy Ion Radiation Biology and Medicine, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000, China
| | - Zhengjia Zhao
- State Key Laboratory of Applied Organic Chemistry and College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China
| | - Baoxin Zhang
- State Key Laboratory of Applied Organic Chemistry and College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China
| | - Jianguo Fang
- State Key Laboratory of Applied Organic Chemistry and College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China.,School of Chemistry and Chemical Engineering, Nanjing University of Science & Technology, Nanjing, Jiangsu 210094, China
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3
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Sharapov MG, Gudkov SV, Lankin VZ, Novoselov VI. Role of Glutathione Peroxidases and Peroxiredoxins in Free Radical-Induced Pathologies. BIOCHEMISTRY. BIOKHIMIIA 2021; 86:1418-1433. [PMID: 34906041 DOI: 10.1134/s0006297921110067] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
In this review, we discuss the pathogenesis of some socially significant diseases associated with the development of oxidative stress, such as atherosclerosis, diabetes, and radiation sickness, as well as the possibilities of the therapeutic application of low-molecular-weight natural and synthetic antioxidants for the correction of free radical-induced pathologies. The main focus of this review is the role of two phylogenetically close families of hydroperoxide-reducing antioxidant enzymes peroxiredoxins and glutathione peroxidases - in counteracting oxidative stress. We also present examples of the application of exogenous recombinant antioxidant enzymes as therapeutic agents in the treatment of pathologies associated with free-radical processes and discuss the prospects of the therapeutic use of exogenous antioxidant enzymes, as well as the ways to improve their therapeutic properties.
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Affiliation(s)
- Mars G Sharapov
- Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region, 142290, Russia.
| | - Sergey V Gudkov
- Prokhorov Institute of General Physics, Russian Academy of Sciences, Moscow, 119991, Russia.,Institute of Biology and Biomedicine, Lobachevsky State University of Nizhni Novgorod, Nizhny Novgorod, 603022, Russia.,All-Russian Research Institute of Phytopathology, Bolshiye Vyazemy, 143050, Russia
| | - Vadim Z Lankin
- National Medical Research Center of Cardiology, Ministry of Health of Russia, Moscow, 121552, Russia
| | - Vladimir I Novoselov
- Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region, 142290, Russia
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4
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Sharapov MG, Glushkova OV, Parfenyuk SB, Gudkov SV, Lunin SM, Novoselova EG. The role of TLR4/NF-κB signaling in the radioprotective effects of exogenous Prdx6. Arch Biochem Biophys 2021; 702:108830. [PMID: 33727039 DOI: 10.1016/j.abb.2021.108830] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/14/2021] [Revised: 02/26/2021] [Accepted: 03/07/2021] [Indexed: 01/11/2023]
Abstract
Peroxiredoxin 6 (Prdx6) is a bifunctional enzyme with multi-substrate peroxidase and phospholipase activities that is involved in cell redox homeostasis and regulates intracellular processes. Previously, recombinant Prdx6 was shown to exert a radioprotective effect during whole-body exposure to a lethal dose of X-ray radiation. Moreover, a mutant form Prdx6-C47S, which lacks peroxidase activity, also had a radioprotective effect, and this indicates that the mechanism of radioprotection is unknown. The present study was aimed to test the hypothesis that the radioprotective effect of Prdx6 and Prdx6-C47S may be mediated through the TLR4/NF-κB signaling pathway. It was demonstrated that exogenously applied Prdx6 protected 3T3 fibroblast cells against LD50 X-ray radiation in vitro. Pretreatment with Prdx6 increased cell survival, stimulated proliferation, normalized the level of reactive oxygen species in culture, and suppressed apoptosis and necrosis. Wild-type Prdx6 and, to a lesser degree, the Prdx6-C47S mutant proteins promoted a significant increase in NF-κB activation in irradiated cells, which likely contributes to the antiapoptotic effect. Pretreatment with TLR4 inhibitors, especially those directed to the extracellular part of the receptor, significantly reduced the radioprotective effect, and this supports the role of TLR4 signaling in the protective effects of Prdx6. Therefore, the radioprotective effect of Prdx6 was related not only to its antioxidant properties, but also to its ability to trigger cellular defense mechanisms through interaction with the TLR4 receptor and subsequent activation of the NF-κB pathway. Recombinant Prdx6 may be useful for the development of a new class of safe radioprotective compounds that have a combination of antioxidant and immunomodulatory properties.
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Affiliation(s)
- Mars G Sharapov
- Institute of Cell Biophysics of the Russian Academy of Sciences, PSCBR RAS, Pushchino, Russia.
| | - Olga V Glushkova
- Institute of Cell Biophysics of the Russian Academy of Sciences, PSCBR RAS, Pushchino, Russia
| | - Svetlana B Parfenyuk
- Institute of Cell Biophysics of the Russian Academy of Sciences, PSCBR RAS, Pushchino, Russia
| | - Sergey V Gudkov
- Prokhorov General Physics Institute of the Russian Academy of Sciences, Moscow, Russia
| | - Sergey M Lunin
- Institute of Cell Biophysics of the Russian Academy of Sciences, PSCBR RAS, Pushchino, Russia
| | - Elena G Novoselova
- Institute of Cell Biophysics of the Russian Academy of Sciences, PSCBR RAS, Pushchino, Russia
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5
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Riviere E, Rossi SP, Tavalieri YE, Muñoz de Toro MM, Ponzio R, Puigdomenech E, Levalle O, Martinez G, Terradas C, Calandra RS, Matzkin ME, Frungieri MB. Melatonin daily oral supplementation attenuates inflammation and oxidative stress in testes of men with altered spermatogenesis of unknown aetiology. Mol Cell Endocrinol 2020; 515:110889. [PMID: 32622722 DOI: 10.1016/j.mce.2020.110889] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/24/2020] [Revised: 05/16/2020] [Accepted: 06/01/2020] [Indexed: 12/20/2022]
Abstract
We have previously shown an inverse correlation between testicular melatonin concentration and inflammation/oxidative stress-related markers levels in infertile men showing unexplained azoospermia. Here, we evaluated the impact of melatonin oral supplementation (daily 3 mg dose used to treat sleep disorders) in the incidence of local inflammation, oxidative stress, and tubular wall fibrosis development in young and middle-aged infertile adult men. Compared with testes without histological alterations, gonads with morphological abnormalities showed lower melatonin concentration along with increased macrophage numbers, TBARS generation, and expression levels of inflammation-related markers and antioxidant enzymes, as well as tubular wall collagen fibers disorganization and thickening. Melatonin oral supplementation not only increased its own testicular levels but also decreased inflammation- and oxidative stress-related markers levels, and improved the tubular wall aspect. Overall, our work provides insights into the potential benefits of melatonin on the inflammatory and oxidative status in testes of patients suffering from unexplained infertility.
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Affiliation(s)
- Eugenia Riviere
- Instituto de Biología y Medicina Experimental, CONICET, Ciudad de Buenos Aires, C1428ADN, Argentina; Cátedra de Química, Ciclo Básico Común, Ciudad de Buenos Aires, C1405CAE, Argentina
| | - Soledad P Rossi
- Instituto de Biología y Medicina Experimental, CONICET, Ciudad de Buenos Aires, C1428ADN, Argentina; Cátedra de Bioquímica Humana, Facultad de Medicina, Universidad de Buenos Aires, Ciudad de Buenos Aires, C1121ABG, Argentina
| | - Yamil E Tavalieri
- Instituto de Salud y Ambiente del Litoral, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, 3000, Argentina
| | - Mónica M Muñoz de Toro
- Instituto de Salud y Ambiente del Litoral, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, 3000, Argentina
| | - Roberto Ponzio
- Instituto de Investigaciones en Reproducción, Facultad de Medicina, Universidad de Buenos Aires, Ciudad de Buenos Aires, C1121ABG, Argentina
| | | | - Oscar Levalle
- División Endocrinología, Hospital Durand, Facultad de Medicina, Universidad de Buenos Aires, Ciudad de Buenos Aires, C1405DCS, Argentina
| | | | - Claudio Terradas
- Instituto Médico PREFER, San Martín, Buenos Aires, B1650, Argentina; División Endocrinología, Hospital Durand, Facultad de Medicina, Universidad de Buenos Aires, Ciudad de Buenos Aires, C1405DCS, Argentina; Fertilidad San Isidro, Buenos Aires, B1642, Argentina
| | - Ricardo S Calandra
- Instituto de Biología y Medicina Experimental, CONICET, Ciudad de Buenos Aires, C1428ADN, Argentina
| | - María E Matzkin
- Instituto de Biología y Medicina Experimental, CONICET, Ciudad de Buenos Aires, C1428ADN, Argentina; Cátedra de Química, Ciclo Básico Común, Ciudad de Buenos Aires, C1405CAE, Argentina.
| | - Mónica B Frungieri
- Instituto de Biología y Medicina Experimental, CONICET, Ciudad de Buenos Aires, C1428ADN, Argentina; Cátedra de Química, Ciclo Básico Común, Ciudad de Buenos Aires, C1405CAE, Argentina
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6
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Soliman MM, Elshazly SA, Aldhahrani A. Gamma-irradiation-induced testicular oxidative stress and apoptosis: Mitigation by l-carnitine. J Biochem Mol Toxicol 2020; 34:e22565. [PMID: 32645248 DOI: 10.1002/jbt.22565] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/25/2020] [Revised: 05/15/2020] [Accepted: 06/17/2020] [Indexed: 01/01/2023]
Abstract
The current study evaluated the potential ameliorative and protective impacts of l-carnitine (L-CAR) against γ-irradiation (RAD)-induced oxidative stress and apoptosis in mice testes. Male Swiss mice were allocated into four groups (n = 7). Group 1 served as a control that received saline intraperitoneally (IP). Group 2 received L-CAR (10 mg/kg bw/day; IP in saline) for 17 days. Group 3 received saline for 17 days and on day 7 exposed to RAD at a dose of 0.1 Gy per day for consecutive 10 days. Group 4 (L-CAR + RAD), received L-CAR same as in group 2 and on day 7 exposed to RAD for consecutive 10 days. Testicular antioxidants (malondialdehyde, MDA; γ-glutamyl-cysteine synthetase, gGCS; and catalase) were altered by γ-irradiation. Preadministration of L-CAR protected γ-irradiated mice from altered changes induced by γ-irradiation. γ-Irradiation affected the mRNA expression of pro-apoptotic, apoptotic, and anti-apoptotic genes (c-jun, c-fos, Bcl-xl, caspase-3, and BAX). All altered genes were ameliorated by prior l-carnitine administration to γ-irradiated mice. Testicular cells showed deformities and edema with congestion in seminiferous tubules and strong immunoreactivity for caspase-9 and a decrease in immunoreactivity of Bcl-2 in histological and immunohistochemical examination. Prior administration of L-CAR to γ-irradiated mice protected this group from reported changes in caspase-9 and Bcl-2 immunostaining. In conclusion, the current study provides evidence for the protective and ameliorative impacts of L-CAR against γ-irradiation-induced testicular oxidative stress and apoptosis at biochemical, molecular, and cellular levels.
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Affiliation(s)
- Mohamed M Soliman
- Clinical Laboratory Sciences, Turabah University College, Taif University, Turabah, Saudi Arabia.,Biochemistry Department, Faculty of Veterinary Medicine, Benha University, Benha, Egypt
| | - Samir A Elshazly
- Biochemistry Department, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafrelsheikh, Egypt
| | - Adil Aldhahrani
- Clinical Laboratory Sciences, Turabah University College, Taif University, Turabah, Saudi Arabia
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7
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Molecular Cloning, Expression, and Function of Synechocystis PCC6803 Type II Peroxiredoxin (sll1621) Gene in Escherichia coli Cells under Salinity Stress Conditions. JOURNAL OF PURE AND APPLIED MICROBIOLOGY 2020. [DOI: 10.22207/jpam.14.2.14] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
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8
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Arato I, Grande G, Barrachina F, Bellucci C, Lilli C, Jodar M, Aglietti MC, Mancini F, Vincenzoni F, Pontecorvi A, Calafiore R, Oliva R, Luca G, Mancuso F, Milardi D. " In vitro" Effect of Different Follicle-Stimulating Hormone Preparations on Sertoli Cells: Toward a Personalized Treatment for Male Infertility. Front Endocrinol (Lausanne) 2020; 11:401. [PMID: 32625170 PMCID: PMC7314925 DOI: 10.3389/fendo.2020.00401] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/23/2020] [Accepted: 05/19/2020] [Indexed: 01/02/2023] Open
Abstract
Follicle-stimulating hormone (FSH), a major regulator of spermatogenesis, has a crucial function in the development and function of the testis and it is extensively given as a fertility treatment to stimulate spermatogenesis. We analyzed the effects of different FSH preparations (α-follitropin, β-follitropin, and urofollitropin) in combination with testosterone on porcine pre-pubertal Sertoli cells. To study the effect of the different FSH treatments in the Sertoli cell function we performed Real Time PCR analysis of AMH, inhibin B, and FSH-r, an ELISA assay for AMH and inhibin B, and a high-throughput comparative proteomic analysis. We verified that all three preparations induced a reduction of AMH in terms of mRNA and secreted proteins, and an increase of inhibin B in terms of mRNA in all the FSH formulations, while solely α-follitropin produced an increase of secreted inhibin B in the culture medium. Comparative proteomic analysis of the three FSH preparations identified 46 proteins, 11 up-regulated and 2 down-regulated. Surprisingly, the combination of testosterone with β-follitropin specifically induced an up-regulation of eight specific secreted proteins. Our study, showing that the three different FSH preparations induce different effects, could offer the opportunity to shed light inside new applications to a personalized reproductive medicine.
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Affiliation(s)
- Iva Arato
- Department of Experimental Medicine, University of Perugia, Perugia, Italy
| | - Giuseppe Grande
- Research Unit on Human Reproduction, International Scientific Institute Paul VI, Rome, Italy
- Division of Endocrinology, Fondazione Policlinico Universitario “Agostino Gemelli”, Rome, Italy
| | - Ferran Barrachina
- Molecular Biology of Reproduction and Development Research Group, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
| | - Catia Bellucci
- Department of Experimental Medicine, University of Perugia, Perugia, Italy
| | - Cinzia Lilli
- Department of Experimental Medicine, University of Perugia, Perugia, Italy
| | - Meritxell Jodar
- Molecular Biology of Reproduction and Development Research Group, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
- Biochemistry and Molecular Genetics Service, Hospital Clínic, Barcelona, Spain
| | | | - Francesca Mancini
- Research Unit on Human Reproduction, International Scientific Institute Paul VI, Rome, Italy
| | - Federica Vincenzoni
- Biochemistry and Clinical Biochemistry Institute, School of Medicine, Catholic University of Rome, Rome, Italy
- Department of Laboratory Diagnostic and Infectious Diseases, Fondazione Policlinico “A. Gemelli” IRCCS, Rome, Italy
| | - Alfredo Pontecorvi
- Research Unit on Human Reproduction, International Scientific Institute Paul VI, Rome, Italy
- Division of Endocrinology, Fondazione Policlinico Universitario “Agostino Gemelli”, Rome, Italy
| | - Riccardo Calafiore
- Department of Medicine, University of Perugia, Perugia, Italy
- Division of Medical Andrology and Endocrinology of Reproduction, University of Perugia and Saint Mary Hospital, Terni, Italy
| | - Rafael Oliva
- Molecular Biology of Reproduction and Development Research Group, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain
- Biochemistry and Molecular Genetics Service, Hospital Clínic, Barcelona, Spain
| | - Giovanni Luca
- Department of Experimental Medicine, University of Perugia, Perugia, Italy
- Division of Medical Andrology and Endocrinology of Reproduction, University of Perugia and Saint Mary Hospital, Terni, Italy
- *Correspondence: Giovanni Luca
| | - Francesca Mancuso
- Department of Experimental Medicine, University of Perugia, Perugia, Italy
| | - Domenico Milardi
- Research Unit on Human Reproduction, International Scientific Institute Paul VI, Rome, Italy
- Division of Endocrinology, Fondazione Policlinico Universitario “Agostino Gemelli”, Rome, Italy
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Mohammadi S, Kianmehr M, Mohammadi M, Fahimian Z, Karimimanesh E, Farazifar M, Nakhaei Z, Golamneghad N, Bolourifard B, Gholamin M, Mansouri A, Mahmoodi M RS, Soleymanifard S, Boroumand-Noughabi S, Ghandy N, Delshad A, Mohammadzadeh F, Norasteh H, Ghayour-Mobarhan M, Ferns GA. Correlation between expression of CatSper1,2 and sperm parameters in the gamma irradiated adult mouse testis. Int J Radiat Biol 2019; 95:691-696. [PMID: 30821561 DOI: 10.1080/09553002.2019.1552372] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Abstract
Propose: CatSper protein channels are responsible for the entry of Ca2+ into sperm cells. These proteins play an important role in motility and male fertility. So it is important to find out whether or not environmental factors, such as gamma radiation, have an effect on the expression of Catsper genes. In this study, we investigated the effects of gamma radiation on the expression of CatSper1 and CatSper2 genes. Materials and methods: Twenty-one male NMRI mice were divided into three groups: a control group without gamma radiation, and two experimental groups; Group 1 treated with 1 Gy of gamma radiation, and Group 2 treated with a higher dose of 2 Gy gamma radiation. Testes were removed from all groups of animals 35 days following irradiation and the testicular tissue, processed and embedded in paraffin blocks for sectioning and histological examination. Sperm samples were also taken from the epididymis for microscopic. Sperm parameters such as sperm count, morphology, motility, and viability rates were analyzed. Expression of CatSper genes was evaluated using Real-time PCR. Data were analyzed using the SPSS software and ANOVA test. Results: Our results showed that after treatment with gamma radiation, testes morphology was changed. Epididymal sperm count, motility, and morphology rates were significantly affected in both experimental groups compared to the control group. The relative expressions of CatSper 1 and 2 genes were significantly reduced in the irradiated mice (1 Gy and 2 Gy) than non-irradiated ones. Conclusions: Gamma radiations not only change testes histology and sperm parameters, but also decrease the expression of CatSper 1 and 2 genes in male mice.
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Affiliation(s)
- Shabnam Mohammadi
- a Neurogenic Inflammation Research Center , Mashhad University of Medical Sciences , Mashhad , Iran
| | - Mojtaba Kianmehr
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Maryam Mohammadi
- c Department of Public Health , School of Health, Shahid Beheshti University of Medical Sciences , Tehran , Iran
| | - Zahra Fahimian
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Elham Karimimanesh
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Mostafa Farazifar
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Zahra Nakhaei
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Nafiseh Golamneghad
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Basir Bolourifard
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Mehran Gholamin
- d Department of Laboratory Sciences , School of Paramedical Sciences, Mashhad University of Medical Sciences , Mashhad , Iran
| | - Atena Mansouri
- e Student Research Committee, Nanotechnology Research Center , School of Pharmacy, Mashhad University of Medical Sciences , Mashhad , Iran
| | | | - Shokouhozaman Soleymanifard
- g Department of Medical Physics , School of Medicine, Mashhad University of Medical Sciences , Mashhad , Iran
| | | | - Nasibeh Ghandy
- a Neurogenic Inflammation Research Center , Mashhad University of Medical Sciences , Mashhad , Iran
| | - Ali Delshad
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Fatemeh Mohammadzadeh
- b Student Research Committee, Faculty of Medicine , Gonabad University of Medical Sciences , Gonabad , Iran
| | - Hojjat Norasteh
- a Neurogenic Inflammation Research Center , Mashhad University of Medical Sciences , Mashhad , Iran
| | - Majid Ghayour-Mobarhan
- i Biochemistry of Nutritional Research Center , Mashhad University of Medical Science , Mashhad , Iran
| | - Gordon Aa Ferns
- j Department of Medical Education , Brighton & Sussex Medical School, Falmer , Brighton , UK
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10
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Qu N, Itoh M, Sakabe K. Effects of Chemotherapy and Radiotherapy on Spermatogenesis: The Role of Testicular Immunology. Int J Mol Sci 2019; 20:E957. [PMID: 30813253 PMCID: PMC6413003 DOI: 10.3390/ijms20040957] [Citation(s) in RCA: 49] [Impact Index Per Article: 8.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2019] [Revised: 02/20/2019] [Accepted: 02/20/2019] [Indexed: 02/06/2023] Open
Abstract
Substantial improvements in cancer treatment have resulted in longer survival and increased quality of life in cancer survivors with minimized long-term toxicity. However, infertility and gonadal dysfunction continue to be recognized as adverse effects of anticancer therapy. In particular, alkylating agents and irradiation induce testicular damage that results in prolonged azoospermia. Although damage to and recovery of spermatogenesis after cancer treatment have been extensively studied, there is little information regarding the role of differences in testicular immunology in cancer treatment-induced male infertility. In this review, we briefly summarize available rodent and human data on immunological differences in chemotherapy or radiotherapy.
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Affiliation(s)
- Ning Qu
- Department of Anatomy, Division of Basic Medical Science, Tokai University School of Medicine, Kanagawa 259-1193, Japan.
- Department of Anatomy, Tokyo Medical University, Tokyo 160-8402, Japan.
| | - Masahiro Itoh
- Department of Anatomy, Tokyo Medical University, Tokyo 160-8402, Japan.
| | - Kou Sakabe
- Department of Anatomy, Division of Basic Medical Science, Tokai University School of Medicine, Kanagawa 259-1193, Japan.
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11
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Baselet B, Sonveaux P, Baatout S, Aerts A. Pathological effects of ionizing radiation: endothelial activation and dysfunction. Cell Mol Life Sci 2019; 76:699-728. [PMID: 30377700 PMCID: PMC6514067 DOI: 10.1007/s00018-018-2956-z] [Citation(s) in RCA: 157] [Impact Index Per Article: 26.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/11/2018] [Revised: 10/19/2018] [Accepted: 10/23/2018] [Indexed: 01/13/2023]
Abstract
The endothelium, a tissue that forms a single layer of cells lining various organs and cavities of the body, especially the heart and blood as well as lymphatic vessels, plays a complex role in vascular biology. It contributes to key aspects of vascular homeostasis and is also involved in pathophysiological processes, such as thrombosis, inflammation, and hypertension. Epidemiological data show that high doses of ionizing radiation lead to cardiovascular disease over time. The aim of this review is to summarize the current knowledge on endothelial cell activation and dysfunction after ionizing radiation exposure as a central feature preceding the development of cardiovascular diseases.
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Affiliation(s)
- Bjorn Baselet
- Radiobiology Unit, Belgian Nuclear Research Centre (SCK•CEN), Mol, Belgium
- Institute of Experimental and Clinical Research (IREC), Pole of Pharmacology and Therapeutics, Université catholique de Louvain (UCL), Brussels, Belgium
| | - Pierre Sonveaux
- Institute of Experimental and Clinical Research (IREC), Pole of Pharmacology and Therapeutics, Université catholique de Louvain (UCL), Brussels, Belgium
| | - Sarah Baatout
- Radiobiology Unit, Belgian Nuclear Research Centre (SCK•CEN), Mol, Belgium
- Department of Molecular Biotechnology, Ghent University, Ghent, Belgium
| | - An Aerts
- Radiobiology Unit, Belgian Nuclear Research Centre (SCK•CEN), Mol, Belgium.
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12
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Sharapov MG, Novoselov VI, Gudkov SV. Radioprotective Role of Peroxiredoxin 6. Antioxidants (Basel) 2019; 8:E15. [PMID: 30621289 PMCID: PMC6356814 DOI: 10.3390/antiox8010015] [Citation(s) in RCA: 37] [Impact Index Per Article: 6.2] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2018] [Revised: 12/30/2018] [Accepted: 01/01/2019] [Indexed: 02/06/2023] Open
Abstract
Peroxiredoxin 6 (Prdx6) is a member of an evolutionary ancient family of peroxidase enzymes with diverse functions in the cell. Prdx6 is an important enzymatic antioxidant. It reduces a wide range of peroxide substrates in the cell, thus playing a leading role in the maintenance of the redox homeostasis in mammalian cells. Beside peroxidase activity, Prdx6 has been shown to possess an activity of phospholipase A2, an enzyme playing an important role in membrane phospholipid metabolism. Moreover, Prdx6 takes part in intercellular and intracellular signal transduction due to its peroxidase and phospholipase activity, thus facilitating the initiation of regenerative processes in the cell, suppression of apoptosis, and activation of cell proliferation. Being an effective and important antioxidant enzyme, Prdx6 plays an essential role in neutralizing oxidative stress caused by various factors, including action of ionizing radiation. Endogenous Prdx6 has been shown to possess a significant radioprotective potential in cellular and animal models. Moreover, intravenous infusion of recombinant Prdx6 to animals before irradiation at lethal or sublethal doses has shown its high radioprotective effect. Exogenous Prdx6 effectively alleviates the severeness of radiation lesions, providing normalization of the functional state of radiosensitive organs and tissues, and leads to a significant elevation of the survival rate of animals. Prdx6 can be considered as a potent and promising radioprotective agent for reducing the pathological effect of ionizing radiation on mammalian organisms. The radioprotective properties and mechanisms of radioprotective action of Prdx6 are discussed in the current review.
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Affiliation(s)
- Mars G Sharapov
- Laboratory of Mechanisms of Reception, Institute of Cell Biophysics of the Russian Academy of Sciences, 142290 Pushchino, Russia.
| | - Vladimir I Novoselov
- Laboratory of Mechanisms of Reception, Institute of Cell Biophysics of the Russian Academy of Sciences, 142290 Pushchino, Russia.
| | - Sergey V Gudkov
- Wave Research Center, Prokhorov General Physics Institute of the Russian Academy of Sciences, 119991 Moscow, Russia.
- Department of Experimental Clinical Studies, Moscow Regional Research and Clinical Institute (MONIKI), 129110 Moscow, Russia.
- The Institute of Biology and Biomedicine, Lobachevsky State University of Nizhni Novgorod, 603950 Nizhni Novgorod, Russia.
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13
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Forshaw TE, Holmila R, Nelson KJ, Lewis JE, Kemp ML, Tsang AW, Poole LB, Lowther WT, Furdui CM. Peroxiredoxins in Cancer and Response to Radiation Therapies. Antioxidants (Basel) 2019; 8:antiox8010011. [PMID: 30609657 PMCID: PMC6356878 DOI: 10.3390/antiox8010011] [Citation(s) in RCA: 26] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2018] [Revised: 12/23/2018] [Accepted: 12/25/2018] [Indexed: 12/11/2022] Open
Abstract
Peroxiredoxins have a long-established cellular function as regulators of redox metabolism by catalyzing the reduction of peroxides (e.g., H2O2, lipid peroxides) with high catalytic efficiency. This activity is also critical to the initiation and relay of both phosphorylation and redox signaling in a broad range of pathophysiological contexts. Under normal physiological conditions, peroxiredoxins protect normal cells from oxidative damage that could promote oncogenesis (e.g., environmental stressors). In cancer, higher expression level of peroxiredoxins has been associated with both tumor growth and resistance to radiation therapies. However, this relationship between the expression of peroxiredoxins and the response to radiation is not evident from an analysis of data in The Cancer Genome Atlas (TCGA) or NCI60 panel of cancer cell lines. The focus of this review is to summarize the current experimental knowledge implicating this class of proteins in cancer, and to provide a perspective on the value of targeting peroxiredoxins in the management of cancer. Potential biases in the analysis of the TCGA data with respect to radiation resistance are also highlighted.
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Affiliation(s)
- Tom E Forshaw
- Department of Internal Medicine, Section on Molecular Medicine, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.
| | - Reetta Holmila
- Department of Internal Medicine, Section on Molecular Medicine, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.
| | - Kimberly J Nelson
- Department of Biochemistry, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.
| | - Joshua E Lewis
- The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
| | - Melissa L Kemp
- The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
| | - Allen W Tsang
- Department of Internal Medicine, Section on Molecular Medicine, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.
| | - Leslie B Poole
- Department of Biochemistry, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.
| | - W Todd Lowther
- Department of Biochemistry, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.
| | - Cristina M Furdui
- Department of Internal Medicine, Section on Molecular Medicine, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.
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14
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Peroxiredoxins in Colorectal Cancer: Predictive Biomarkers of Radiation Response and Therapeutic Targets to Increase Radiation Sensitivity? Antioxidants (Basel) 2018; 7:antiox7100136. [PMID: 30301137 PMCID: PMC6210826 DOI: 10.3390/antiox7100136] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/03/2018] [Revised: 09/27/2018] [Accepted: 10/03/2018] [Indexed: 02/06/2023] Open
Abstract
Colorectal cancer (CRC) is the third most common cancer in the Western world, with one-third of cases located in the rectum. Preoperative radiotherapy is the standard of care for many patients with rectal cancer but has a highly variable response rate. The ability to predict response would be of great clinical utility. The response of cells to ionizing radiation is known to involve immediate damage to biomolecules and more sustained disruption of redox homeostasis leading to cell death. The peroxiredoxins are an important group of thiol-dependent antioxidants involved in protecting cells from oxidative stress and regulating signaling pathways involved in cellular responses to oxidative stress. All six human peroxiredoxins have shown increased expression in CRC and may be associated with clinicopathological features and tumor response to ionizing radiation. Peroxiredoxins can act as markers of oxidative stress in various biological systems but they have not been investigated in this capacity in CRC. As such, there is currently insufficient evidence to support the role of peroxiredoxins as clinical biomarkers, but it is an area worthy of investigation. Future research should focus on the in vivo response of rectal cancer to radiotherapy and the redox status of peroxiredoxins in rectal cancer cells, in order to predict response to radiotherapy. The peroxiredoxin system is also a potential therapeutic target for CRC.
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15
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Sharapov MG, Fesenko EE, Novoselov VI. The Role of Peroxiredoxins in Various Diseases Caused by Oxidative Stress and the Prospects of Using Exogenous Peroxiredoxins. Biophysics (Nagoya-shi) 2018. [DOI: 10.1134/s0006350918040164] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/26/2022] Open
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16
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Wang J, Li T, Feng J, Li L, Wang R, Cheng H, Yuan Y. Kaempferol protects against gamma radiation-induced mortality and damage via inhibiting oxidative stress and modulating apoptotic molecules in vivo and vitro. ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY 2018; 60:128-137. [PMID: 29705372 DOI: 10.1016/j.etap.2018.04.014] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/28/2017] [Revised: 04/12/2018] [Accepted: 04/16/2018] [Indexed: 05/02/2023]
Abstract
To investigate the potential protective effect of kaempferol, a representative flavonoid, against radiation induced mortality and injury in vivo and vitro.C57BL/6 male mice and human umbilical venous endothelial cells (HUVECs) were pretreated with kaempferol before radiation. We found that kaempferol can effectively increase 30-day survival rate after 8.5 Gy lethal total body irradiation (TBI). Mice were sacrificed at 7th day after 7 Gy TBI, we found kaempferol against radiation-induced tissues damage, by inhibiting the oxidative stress, and attenuating morphological changes and cell apoptosis. In vitro, kaempferol increased HUVECs cell viability and decrease apoptosis. It also mitigated oxidative stress and restored the abnormal expression of prx-5, Cyt-c, Caspase9 and Caspase3 in mRNA and protein level in HUVECs after radiation. Taken together, it suggests kaempferol can protect against gamma-radiation induced tissue damage and mortality. The present study is the first report of the radioprotective role of kaempferol in vivo and vitro.
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Affiliation(s)
- Jing Wang
- Department of Pharmacy, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, 280 Mo He Rd, Shanghai 201999, China; Department of Pharmacy, Punan Hospital, Shanghai 200125, China; Department of Pharmacology, College of Pharmacy, Second Millitary Medical University, Shanghai 200433, China
| | - Tiejun Li
- Department of Pharmacy, Punan Hospital, Shanghai 200125, China
| | - Jingjing Feng
- Department of Pharmacology, College of Pharmacy, Second Millitary Medical University, Shanghai 200433, China
| | - Li Li
- Department of Pharmacology, College of Pharmacy, Second Millitary Medical University, Shanghai 200433, China
| | - Rong Wang
- Department of Pharmacy, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, 280 Mo He Rd, Shanghai 201999, China
| | - Hao Cheng
- Department of Pharmacology, College of Pharmacy, Second Millitary Medical University, Shanghai 200433, China
| | - Yongfang Yuan
- Department of Pharmacy, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, 280 Mo He Rd, Shanghai 201999, China.
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17
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Asadi N, Bahmani M, Kheradmand A, Rafieian-Kopaei M. The Impact of Oxidative Stress on Testicular Function and the Role of Antioxidants in Improving it: A Review. J Clin Diagn Res 2017; 11:IE01-IE05. [PMID: 28658802 DOI: 10.7860/jcdr/2017/23927.9886] [Citation(s) in RCA: 115] [Impact Index Per Article: 14.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/03/2016] [Accepted: 10/25/2016] [Indexed: 01/10/2023]
Abstract
Oxidative stress is an important factor for development of male infertility because of very high rate of cell division and mitochondrial oxygen consumption in testicular tissue as well as comparably higher levels of unsaturated fatty acids in this tissue than in other tissues. Moreover, the level of oxygen pressure is low due to the weakness of testicular artery; therefore, there is a severe cell competition for oxygen. Therefore, the testicular tissue and male reproductive system are particularly susceptible to oxidative stress. On the other hand, exposure to X-ray, toxins and chemicals found in the environment as well as specific physical conditions such as varicocele can exacerbate the oxidative stress and induce apoptosis of germ cells and subsequently spermatogenesis. However, under normal conditions, the body's capacity to produce antioxidants for inhibiting adverse effects of oxidative stress is affected by metabolic process and genetic structure. Besides that, environmental factors such as diet, pollutants, and chemicals can affect this capacity. Thus, the body's antioxidant system alone is not able to neutralize all free radicals and prevent harmful complications of oxidative stress. Therefore, use of antioxidants and development of antioxidant therapy can break down the oxidative chain reaction and play a very significant role in increasing the body's capacity to fight free radical-induced oxidative stress, and therefore improve the process of spermatogenesis.
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Affiliation(s)
- Nematollah Asadi
- Student Research Committee of Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran: Biotechnology laboratory of ASRI, Karaj, Iran
| | - Mahmoud Bahmani
- Leishmaniasis Research Center, Ilam University of Medical Sciences, Ilam, Iran
| | - Arash Kheradmand
- Razi Herbal Medicines Research center, Lorestan University of Medical Sciences, Khorramabad, Iran
| | - Mahmoud Rafieian-Kopaei
- Medical Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
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18
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Selvaratnam JS, Robaire B. Effects of Aging and Oxidative Stress on Spermatozoa of Superoxide-Dismutase 1- and Catalase-Null Mice. Biol Reprod 2016; 95:60. [PMID: 27465136 PMCID: PMC5333935 DOI: 10.1095/biolreprod.116.141671] [Citation(s) in RCA: 47] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/05/2016] [Accepted: 07/22/2016] [Indexed: 12/28/2022] Open
Abstract
Advanced paternal age is linked to complications in pregnancy and genetic diseases in offspring. Aging results in excess reactive oxygen species (ROS) and DNA damage in spermatozoa; this damage can be transmitted to progeny with detrimental consequences. Although there is a loss of antioxidants with aging, the impact on aging male germ cells of the complete absence of either catalase (CAT) or superoxide dismutase 1 (SOD1) has not been investigated. We used CAT-null (Cat(-/-)) and SOD1-null (Sod(-/-)) mice to determine whether loss of these antioxidants increases germ cell susceptibility to redox dysfunction with aging. Aging reduced fertility and the numbers of Sertoli and germ cells in all mice. Aged Sod(-/-) mice displayed an increased loss of fertility compared to aged wild-type mice. Treatment with the pro-oxidant SIN-10 increased ROS in spermatocytes of aged wild-type and Sod(-/-) mice, while aged Cat(-/-) mice were able to neutralize this ROS. The antioxidant peroxiredoxin 1 (PRDX1) increased with age in wild-type and Cat(-/-) mice but was consistently low in young and aged Sod(-/-) mice. DNA damage and repair markers (γ-H2AX and 53BP1) were reduced with aging and lower in young Sod(-/-) and Cat(-/-) mice. Colocalization of γ-H2AX and 53BP1 suggested active repair in young wild-type mice but reduced in young Cat(-/-) and in Sod(-/-) mice and with age. Oxidative DNA damage (8-oxodG) increased in young Sod(-/-) mice and with age in all mice. These studies show that aged Sod(-/-) mice display severe redox dysfunction, while wild-type and Cat(-/-) mice have compensatory mechanisms to partially alleviate oxidative stress and reduce age-related DNA damage in spermatozoa. Thus, SOD1 but not CAT is critical to the maintenance of germ cell quality with aging.
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Affiliation(s)
- Johanna S Selvaratnam
- Department of Pharmacology and Therapeutics, McGill University, Montréal, Québec, Canada
| | - Bernard Robaire
- Department of Pharmacology and Therapeutics, McGill University, Montréal, Québec, Canada Department of Obstetrics and Gynecology, McGill University, Montréal, Québec, Canada
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19
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Orhon ZN, Uzal C, Kanter M, Erboga M, Demiroglu M. Protective effects of Nigella sativa on gamma radiation-induced jejunal mucosal damage in rats. Pathol Res Pract 2016; 212:437-43. [DOI: 10.1016/j.prp.2016.02.017] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/01/2015] [Revised: 02/04/2016] [Accepted: 02/15/2016] [Indexed: 10/22/2022]
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20
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Ko EY, Sabanegh ES, Agarwal A. Male infertility testing: reactive oxygen species and antioxidant capacity. Fertil Steril 2014; 102:1518-27. [DOI: 10.1016/j.fertnstert.2014.10.020] [Citation(s) in RCA: 159] [Impact Index Per Article: 14.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/30/2014] [Revised: 10/14/2014] [Accepted: 10/14/2014] [Indexed: 02/07/2023]
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21
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Identification and characterization of germ cell genes expressed in the F9 testicular teratoma stem cell line. PLoS One 2014; 9:e103837. [PMID: 25153150 PMCID: PMC4143169 DOI: 10.1371/journal.pone.0103837] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2014] [Accepted: 07/02/2014] [Indexed: 02/06/2023] Open
Abstract
The F9 cell line, which was derived from a mouse testicular teratoma that originated from pluripotent germ cells, has been used as a model for differentiation. However, it is largely unknown whether F9 cells possess the characteristics of male germ cells. In the present study, we investigated spermatogenic stage- and cell type-specific gene expression in F9 cells. Analysis of previous microarray data showed that a large number of stage-regulated germ cell genes are expressed in F9 cells. Specifically, genes that are prominently expressed in spermatogonia and have transcriptional regulatory functions appear to be enriched in F9 cells. Our in silico and in vitro analyses identified several germ cell-specific or -predominant genes that are expressed in F9 cells. Among them, strong promoter activities were observed in the regions upstream of the spermatogonial genes, Dmrt1 (doublesex and mab-3 related transcription factor 1), Stra8 (stimulated by retinoic acid gene 8) and Tex13 (testis expressed gene 13), in F9 cells. A detailed analysis of the Tex13 promoter allowed us to identify an enhancer and a region that is implicated in germ cell-specificity. We also found that Tex13 expression is regulated by DNA methylation. Finally, analysis of GFP (green fluorescent protein) TEX13 localization revealed that the protein distributes heterogeneously in the cytoplasm and nucleus, suggesting that TEX13 shuttles between these two compartments. Taken together, our results demonstrate that F9 cells express numerous spermatogonial genes and could be used for transcriptional studies focusing on such genes. As an example of this, we use F9 cells to provide comprehensive expressional information about Tex13, and report that this gene appears to encode a germ cell-specific protein that functions in the nucleus during early spermatogenesis.
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22
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23
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Reisz JA, Bansal N, Qian J, Zhao W, Furdui CM. Effects of ionizing radiation on biological molecules--mechanisms of damage and emerging methods of detection. Antioxid Redox Signal 2014; 21:260-92. [PMID: 24382094 PMCID: PMC4060780 DOI: 10.1089/ars.2013.5489] [Citation(s) in RCA: 477] [Impact Index Per Article: 43.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/19/2013] [Revised: 12/07/2013] [Accepted: 01/01/2014] [Indexed: 12/13/2022]
Abstract
SIGNIFICANCE The detrimental effects of ionizing radiation (IR) involve a highly orchestrated series of events that are amplified by endogenous signaling and culminating in oxidative damage to DNA, lipids, proteins, and many metabolites. Despite the global impact of IR, the molecular mechanisms underlying tissue damage reveal that many biomolecules are chemoselectively modified by IR. RECENT ADVANCES The development of high-throughput "omics" technologies for mapping DNA and protein modifications have revolutionized the study of IR effects on biological systems. Studies in cells, tissues, and biological fluids are used to identify molecular features or biomarkers of IR exposure and response and the molecular mechanisms that regulate their expression or synthesis. CRITICAL ISSUES In this review, chemical mechanisms are described for IR-induced modifications of biomolecules along with methods for their detection. Included with the detection methods are crucial experimental considerations and caveats for their use. Additional factors critical to the cellular response to radiation, including alterations in protein expression, metabolomics, and epigenetic factors, are also discussed. FUTURE DIRECTIONS Throughout the review, the synergy of combined "omics" technologies such as genomics and epigenomics, proteomics, and metabolomics is highlighted. These are anticipated to lead to new hypotheses to understand IR effects on biological systems and improve IR-based therapies.
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Affiliation(s)
- Julie A Reisz
- Section on Molecular Medicine, Department of Internal Medicine, Wake Forest School of Medicine , Winston-Salem, North Carolina
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24
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Liu J, Rong CT, Li Y, Liu XX, Wang WT, Li N. Vasectomy Induces Oxidative Stress and Up-Regulates the Expression of Peroxiredoxins in Mouse Testis in Short and Early Periods after Surgery. J Urol 2014; 191:1920-6. [PMID: 24440237 DOI: 10.1016/j.juro.2014.01.007] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 01/06/2014] [Indexed: 10/25/2022]
Affiliation(s)
- Juan Liu
- Central Laboratory, Yantai Yu-Huang-Ding Hospital, Yantai, Shandong Province, People's Republic of China
| | - Cheng T. Rong
- Department of Pharmacy, Fuyang People's Hospital, Fuyang, Anhui Province, People's Republic of China
| | - Yan Li
- College of Life Science, Yantai University, Yantai, Shandong Province, People's Republic of China
| | - Xue X. Liu
- Central Laboratory, Yantai Yu-Huang-Ding Hospital, Yantai, Shandong Province, People's Republic of China
| | - Wen T. Wang
- Central Laboratory, Yantai Yu-Huang-Ding Hospital, Yantai, Shandong Province, People's Republic of China
| | - Ning Li
- Central Laboratory, Yantai Yu-Huang-Ding Hospital, Yantai, Shandong Province, People's Republic of China
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25
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Rossi SP, Windschuettl S, Matzkin ME, Terradas C, Ponzio R, Puigdomenech E, Levalle O, Calandra RS, Mayerhofer A, Frungieri MB. Melatonin in testes of infertile men: evidence for anti-proliferative and anti-oxidant effects on local macrophage and mast cell populations. Andrology 2014; 2:436-49. [PMID: 24659586 DOI: 10.1111/j.2047-2927.2014.00207.x] [Citation(s) in RCA: 54] [Impact Index Per Article: 4.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/05/2013] [Revised: 02/11/2014] [Accepted: 02/16/2014] [Indexed: 11/28/2022]
Abstract
Melatonin acting through the hypothalamus and pituitary regulates testicular function. In addition, direct actions of melatonin at the testicular level have been recently suggested. We have described that melatonin inhibits androgen production in hamster Leydig cells via melatonin subtype 1a (mel1a) receptors and the local corticotrophin-releasing hormone (CRH) system. The initial events of the melatonin/CRH signalling pathway have also been established. Melatonin and all components of the melatonergic/CRH system were also detected in Leydig cells of infertile men. This study attempted to search for additional targets of melatonin in the human testis, and to investigate the effects of melatonin on proliferation and the oxidative state in these novel target cells. To this aim, evaluation of human testicular biopsies of patients suffering from hypospermatogenesis or Sertoli cell only syndrome and cell culture studies were performed. Melatonergic receptors were found in macrophages (MACs) and mast cells (MCs) of the human testis. In biopsies of patients suffering idiopathic infertility, melatonin testicular concentrations were negatively correlated with MAC number per mm(2) and TNFα, IL1β and COX2 expression, but positively correlated with the expression of the anti-oxidant enzymes SOD1, peroxiredoxin 1 and catalase. Melatonin inhibited proliferation and the expression of pro-inflammatory cytokines and cyclooxygenase 2 (COX2) in both the human non-testicular THP-1 MAC cell line and primary cell cultures of hamster testicular MACs. In the human HMC-1 MC line, melatonin increased the expression of anti-oxidant enzymes and decreased reactive oxygen species (ROS) generation. The results reveal new testicular targets of melatonin and describe anti-proliferative and anti-inflammatory effects of this hormone on testicular MACs. Furthermore, melatonin might provide protective effects against oxidative stress in testicular MCs.
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Affiliation(s)
- S P Rossi
- Instituto de Biología y Medicina Experimental, Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina; Departamento de Bioquímica Humana, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina
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26
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Krogenæs AK, Ropstad E, Gutleb AC, Hårdnes N, Berg V, Dahl E, Fowler PA. In utero exposure to environmentally relevant concentrations of PCB 153 and PCB 118 disrupts fetal testis development in sheep. JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH. PART A 2014; 77:628-649. [PMID: 24754397 DOI: 10.1080/15287394.2014.887426] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/03/2023]
Abstract
Polychlorinated biphenyls (PCB) are environmental pollutants linked to adverse health effects including endocrine disruption and disturbance of reproductive development. This study aimed to determine whether exposure of pregnant sheep to three different mixtures of PCB 153 and PCB 118 affected fetal testis development. Ewes were treated by oral gavage from mating until euthanasia (d 134), producing three groups of fetuses with distinct adipose tissue PCB levels: high PCB 153/low PCB 118 (n = 13), high PCB 118/low PCB 153 (n = 14), and low PCB 153/low PCB 118 (n = 14). Fetal testes and blood samples were collected for investigation of testosterone, testis morphology, and testis proteome. The body weight of the offspring was lower in the high PCB compared to the low PCB group, but there were no significant differences in testis weight between groups when corrected for body weight. PCB exposure did not markedly affect circulating testosterone. There were no significant differences between groups in number of seminiferous tubules, Sertoli cell only tubules, and ratio between relative areas of seminiferous tubules and interstitium. Two-dimensional (2D) gel-based proteomics was used to screen for proteomic alterations in the high exposed groups relative to low PCB 153/low PCB 118 group. Twenty-six significantly altered spots were identified by liquid chromatography (LC)-mass spectroscopy (MS)/MS. Changes in protein regulation affected cellular processes as stress response, protein synthesis, and cytoskeleton regulation. The study demonstrates that in utero exposure to different environmental relevant PCB mixtures exerted subtle effects on developing fetal testis proteome but did not significantly disturb testis morphology and testosterone production.
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Affiliation(s)
- Anette K Krogenæs
- a Department of Production Animal Sciences , Norwegian School Veterinary Science , Oslo , Norway
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27
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Li J, Yang J, Cheng D, Shen SL, Xiong CL. New clues to identify proteins correlated with Attractin. Andrologia 2013; 46:796-804. [PMID: 23998293 DOI: 10.1111/and.12151] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 06/10/2013] [Indexed: 01/29/2023] Open
Affiliation(s)
- J. Li
- Reproductive Medical Center; Renmin hospital of Wuhan University; Wuhan China
- Family Planning Research Institute; Tongji Medical College; Huazhong University of Science and Technology; Wuhan China
| | - J. Yang
- Reproductive Medical Center; Renmin hospital of Wuhan University; Wuhan China
| | - D. Cheng
- Reproductive Medical Center; Renmin hospital of Wuhan University; Wuhan China
- Family Planning Research Institute; Tongji Medical College; Huazhong University of Science and Technology; Wuhan China
| | - S.-L. Shen
- Department of Pathology; Kindstar Global; Wuhan China
| | - C.-L. Xiong
- Family Planning Research Institute; Tongji Medical College; Huazhong University of Science and Technology; Wuhan China
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28
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Shin HS, Yang WJ, Choi EM. The preventive effect of Se-methylselenocysteine on γ-radiation-induced oxidative stress in rat lungs. J Trace Elem Med Biol 2013. [PMID: 23176811 DOI: 10.1016/j.jtemb.2012.09.003] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
We investigated the preventive effect of Se-methylselenocysteine (MSC) administration on γ-radiation (whole body irradiation, single 10-Gy dose)-induced oxidative damage in rat lungs. Rats were pretreated with MSC (0.75mg/rat/day) for 1 week before γ-irradiation. The MSC pretreatment prevented the irradiation-induced increase in lipid peroxidation and the concomitant decrease in cellular glutathione content. The prevention of irradiation-induced oxidative damage in MSC-pretreated rat lungs appeared to be associated with increased antioxidant capacity, particularly in the glutathione system. The 1-week MSC treatment resulted in an increase in glutathione peroxidase, glutathione reductase, and glucose 6-phosphate dehydrogenase activities, which are involved in glutathione redox cycling. An increase in catalase activity was also observed in the rat lungs. Additionally, a significantly increased level of nuclear factor erythroid 2-related factor 2 (Nrf2) was exhibited in the MSC-treated rat lungs. Heme oxygenase 1, glutathione S-transferase pi, and peroxiredoxin 1, which are known target proteins of Nrf2, were also increased in MSC-treated lungs. These results implicate Nrf2 signaling in the MSC-induced activation of the antioxidant system.
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Affiliation(s)
- Ho-Sang Shin
- Department of Chemistry, University of Incheon, Incheon, Republic of Korea
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Evidence for involvement of cytosolic thioredoxin peroxidase in the excessive resistance of Sf9 Lepidopteran insect cells against radiation-induced apoptosis. PLoS One 2013; 8:e58261. [PMID: 23505474 PMCID: PMC3591413 DOI: 10.1371/journal.pone.0058261] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/13/2012] [Accepted: 02/01/2013] [Indexed: 11/27/2022] Open
Abstract
Lepidopteran insect cells display 50–100 times higher radioresistance compared to human cells, and reportedly have more efficient antioxidant system that can significantly reduce radiation-induced oxidative stress and cell death. However, the antioxidant mechanisms that contribute substantially to this excessive resistance still need to be understood thoroughly. In this study, we investigated the role of thioredoxin peroxidase (TPx) in high-dose γ-radiation response of Sf9 cell line derived from Spodoptera frugiperda, the Fall armyworm. We identified a TPx orthologue (Sf-TPx) in Spodoptera system, with primarily cytosolic localization. Gamma-irradiation at 500 Gy dose significantly up-regulated Sf-TPx, while higher doses (1000 Gy–2000 Gy) had no such effect. G2/M checkpoint induced following 500 Gy was associated with transition of Sf-TPx decamer into enzymatically active dimer. Same effect was observed during G2/M block induced by 5 nM okadaic acid or 10 µM CDK1 (cycline dependent kinase-1) inhibitor roscovitine, thus indicating that radiation-induced Sf-TPx activity is mediated by CDKs. Accumulation of TPx dimer form during G2/M checkpoint might favour higher peroxidase activity facilitating efficient survival at this dose. Confirming this, higher lethal doses (1000 Gy–2000 Gy) caused significantly less accumulation of dimer form and induced dose-dependent apoptosis. A ∼50% knock-down of Sf-TPx by siRNA caused remarkable increase in radiation-induced ROS as well as caspase-3 dependent radiation-induced apoptosis, clearly implying TPx role in the radioresistance of Sf9 cells. Quite importantly, our study demonstrates for the first time that thioredoxin peroxidase contributes significantly in the radioresistance of Lepidopteran Sf9 insect cells, especially in their exemplary resistance against radiation-induced apoptosis. This is an important insight into the antioxidant mechanisms existing in this highly stress-resistant model cell system.
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Pfeifer N, Baston-Büst DM, Hirchenhain J, Friebe-Hoffmann U, Rein DT, Krüssel JS, Hess AP. Selection of the in vitro culture media influences mRNA expression of Hedgehog genes, Il-6, and important genes regarding reactive oxygen species in single murine preimplantation embryos. ScientificWorldJournal 2012; 2012:479315. [PMID: 22919324 PMCID: PMC3417195 DOI: 10.1100/2012/479315] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2012] [Accepted: 04/22/2012] [Indexed: 11/23/2022] Open
Abstract
Background. The aim of this paper was to determine the influence of different in vitro culture media on mRNA expression of Hedgehog genes, il-6, and important genes regarding reactive oxygen species in single mouse embryos. Methods. Reverse transcription of single embryos either cultured in vitro from day 0.5 until 3.5 (COOK's Cleavage medium or Vitrolife's G-1 PLUS medium) or in vivo until day 3.5 post coitum. PCR was carried out for β-actin followed by nested-PCR for shh, ihh, il-6, nox, gpx4, gpx1, and prdx2. Results. The number of murine blastocysts cultured in COOK medium which expressed il-6, gpx4, gpx1, and prdx2 mRNA differed significantly compared to the in vivo group. Except for nox, the mRNA profile of the Vitrolife media group embryos varied significantly from the in vivo ones regarding the number of blastocysts expressing the mRNA of shh, ihh, il-6, gpx4, gpx1 and prdx2. Conclusions. The present study shows that different in vitro culture media lead to different mRNA expression profiles during early development. Even the newly developed in vitro culture media are not able to mimic the female reproductive tract. The question of long-term consequences for children due to assisted reproduction techniques needs to be addressed in larger studies.
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Affiliation(s)
- N Pfeifer
- Department of OB/GYN and REI (UniKiD), Medical Center University of Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany
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Woolston CM, Storr SJ, Ellis IO, Morgan DAL, Martin SG. Expression of thioredoxin system and related peroxiredoxin proteins is associated with clinical outcome in radiotherapy treated early stage breast cancer. Radiother Oncol 2011; 100:308-13. [PMID: 21641069 DOI: 10.1016/j.radonc.2011.05.029] [Citation(s) in RCA: 35] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/24/2010] [Revised: 05/09/2011] [Accepted: 05/12/2011] [Indexed: 10/18/2022]
Abstract
BACKGROUND AND PURPOSE Deregulated redox systems provide cancer cells protection from increased oxidative stress, such as that induced by ionizing radiation. Expression of the thioredoxin system proteins (thioredoxin, thioredoxin reductase and thioredoxin interacting protein) and downstream peroxiredoxins (I-VI), was examined in tumor specimens from early stage breast cancer patients, subsequently treated by breast conserving surgery and locoregional radiotherapy, to determine if redox protein expression is associated with clinical outcome. MATERIAL AND METHODS Nuclear and cytoplasmic expression was assessed using conventional immunohistochemistry on a tissue microarray of 224 tumors. RESULTS High expression of cytoplasmic peroxiredoxin-I correlated with a greater risk of local recurrence (p=0.009). When nuclear and cytoplasmic expression patterns were combined, patients with low nuclear but high cytoplasmic expression of peroxiredoxin-I increased significance (p=0.005). Both were independent factors (p=0.006 and 0.003) from multivariate analysis. Associations were obtained between tumor grade and nuclear thioredoxin interacting protein (p=0.01) and with cytoplasmic expression of peroxiredoxin-V (p=0.007) but not with peroxiredoxin-I suggesting that the latter may exert influence via regulation of oxidative stress rather than via altering the tumor phenotype. CONCLUSIONS Results highlight the potential of using redox protein expression, namely peroxiredoxin-I, to predict clinical outcome and support further studies to validate its usefulness as an independent prognostic, and potentially predictive, marker.
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O'Flaherty C, de Souza AR. Hydrogen peroxide modifies human sperm peroxiredoxins in a dose-dependent manner. Biol Reprod 2010; 84:238-47. [PMID: 20864641 DOI: 10.1095/biolreprod.110.085712] [Citation(s) in RCA: 91] [Impact Index Per Article: 6.1] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022] Open
Abstract
Low levels of reactive oxygen species (ROS) modulate signaling pathways required for human sperm activation, but high levels impair sperm function, leading to infertility. Peroxiredoxins (PRDXs) are enzymes with a dual role as ROS scavengers and modulators of ROS-dependent signaling. The present study aimed to characterize PRDXs in human spermatozoa and possible modifications resulting from hydrogen peroxide (H(2)O(2)). We found PRDX1, PRDX4, PRDX5, and PRDX6 in both seminal plasma and spermatozoa. Using immunocytochemistry, we demonstrated that these PRDXs are differentially localized in the head, acrosome, mitochondrial sheath, and flagellum. These observations were confirmed by immunoblotting using cytosolic, Triton-soluble and -insoluble, and head and flagella sperm fractions. PRDXs are dose-dependently modified by H(2)O(2), as seen by the formation of disulfide bridges and high-molecular-mass complexes. This first study, to our knowledge, on PRDXs in human spermatozoa indicates that PRDX1, PRDX4, PRDX5, and PRDX6 are modified when spermatozoa are challenged with H(2)O(2). This suggests that PRDXs may protect these cells at high levels of H(2)O(2) but could also control H(2)O(2) levels within different cell compartments so that normal sperm activation can occur.
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Affiliation(s)
- Cristian O'Flaherty
- Urology Research Laboratory, Urology Division, Faculty of Medicine, McGill University and Research Institute-McGill University Health Centre, Montréal, Québec, Canada.
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33
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Antiapoptotic effect of l-carnitine on testicular irradiation in rats. J Mol Histol 2010; 41:121-8. [DOI: 10.1007/s10735-010-9267-5] [Citation(s) in RCA: 38] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/24/2010] [Accepted: 04/22/2010] [Indexed: 10/19/2022]
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Wang S, Huang W, Shi H, Lin C, Xie M, Wang J. Localization and expression of peroxiredoxin II in the mouse ovary, oviduct, uterus, and preimplantation embryo. Anat Rec (Hoboken) 2010; 293:291-7. [PMID: 19899112 DOI: 10.1002/ar.21031] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/11/2022]
Abstract
Peroxiredoxin (Prx) II belongs to a recently discovered family of peroxidases that play important roles in antioxidation and signal transduction. In this study, we aimed to study the localization and expression of Prx II in the mouse ovary, oviduct, and uterus, and preimplantation embryos. Immunohistochemical staining analysis showed that, in the ovary, Prx II was expressed in the oocyte cytoplasm of the primary follicle, the secondary follicle, and the premature follicle; Prx II was expressed in germinal vesicle-intact oocytes (GV oocytes) and metaphase II eggs (MII eggs), as well as at various stages in early embryos. Reverse transcription polymerase chain reaction (RT-PCR) results indicated that the Prx II mRNA was expressed at a high level in GV eggs, slightly lower levels in MII eggs, and had no detectable expression in four-cell embryos and early blastocysts. In the oviduct, Prx II was expressed in the epithelia, while in the uterus Prx II was mainly distributed in the endometrial stroma. Taken together, our results suggest that Prx II plays a key antioxidation role in the maturation of oocytes and development of early embryos, thus providing crucial experimental evidence for further exploring the function of Prx II in the development of oocytes and preimplantation embryos.
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Affiliation(s)
- Shie Wang
- Department of Human Anatomy, Histology, and Embryology, Fujian Medical University, Fuzhou, People's Republic of China.
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35
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Zhang B, Wang Y, Su Y. Peroxiredoxins, a novel target in cancer radiotherapy. Cancer Lett 2009; 286:154-60. [PMID: 19500902 DOI: 10.1016/j.canlet.2009.04.043] [Citation(s) in RCA: 95] [Impact Index Per Article: 5.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/24/2009] [Revised: 04/29/2009] [Accepted: 04/30/2009] [Indexed: 01/01/2023]
Abstract
Reactive oxygen species (ROS) are toxic at high levels in the mammalian cells. Mammalian cells have developed many enzymatic and nonenzymatic antioxidative systems in various cellular compartments to maintain an appropriate level of ROS and regulate their action. Peroxiredoxins (Prxs), a family of peroxidase that reduced intracellular peroxides (one type of ROS) with the thioredoxin system as the electron donor, were highly expressed in various cellular compartments. In this minireview, we discussed the regulation of Prxs expression in cancer cell and its relationship with ionizing radiation. As Prxs could be induced by radiation and its expression status could determine the radiosensitivity of cancer cells, Prxs might be a potential target for radiotherapy in cancer.
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Affiliation(s)
- Bo Zhang
- Department of Medical Genetics, Third Military Medical University, Chongqing 400038, China
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Seo JH, Lim JC, Lee DY, Kim KS, Piszczek G, Nam HW, Kim YS, Ahn T, Yun CH, Kim K, Chock PB, Chae HZ. Novel protective mechanism against irreversible hyperoxidation of peroxiredoxin: Nalpha-terminal acetylation of human peroxiredoxin II. J Biol Chem 2009; 284:13455-13465. [PMID: 19286652 DOI: 10.1074/jbc.m900641200] [Citation(s) in RCA: 42] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022] Open
Abstract
Peroxiredoxins (Prxs) are a group of peroxidases containing a cysteine thiol at their catalytic site. During peroxidase catalysis, the catalytic cysteine, referred to as the peroxidatic cysteine (C(P)), cycles between thiol (C(P)-SH) and disulfide (-S-S-) states via a sulfenic (C(P)-SOH) intermediate. Hyperoxidation of the C(P) thiol to its sulfinic (C(P)-SO(2)H) derivative has been shown to be reversible, but its sulfonic (C(P)-SO(3)H) derivative is irreversible. Our comparative study of hyperoxidation and regeneration of Prx I and Prx II in HeLa cells revealed that Prx II is more susceptible than Prx I to hyperoxidation and that the majority of the hyperoxidized Prx II formation is reversible. However, the hyperoxidized Prx I showed much less reversibility because of the formation of its irreversible sulfonic derivative, as verified with C(P)-SO(3)H-specific antiserum. In an attempt to identify the multiple hyperoxidized spots of the Prx I on two-dimensional PAGE analysis, an N-acetylated Prx I was identified as part of the total Prx I using anti-acetylated Lys antibody. Using peptidyl-Asp metalloendopeptidase (EC 3.4.24.33) peptide fingerprints, we found that N(alpha)-terminal acetylation (N(alpha)-Ac) occurred exclusively on Prx II after demethionylation. N(alpha)-Ac of Prx II blocks Prx II from irreversible hyperoxidation without altering its affinity for hydrogen peroxide. A comparative study of non-N(alpha)-acetylated and N(alpha)-terminal acetylated Prx II revealed that N(alpha)-Ac of Prx II induces a significant shift in the circular dichroism spectrum and elevation of T(m) from 59.6 to 70.9 degrees C. These findings suggest that the structural maintenance of Prx II by N(alpha)-Ac may be responsible for preventing its hyperoxidation to form C(P)-SO(3)H.
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Affiliation(s)
- Jae Ho Seo
- School of Biological Sciences and Technology Chonnam National University, Gwangju 500-757, Korea
| | - Jung Chae Lim
- School of Biological Sciences and Technology Chonnam National University, Gwangju 500-757, Korea
| | - Duck-Yeon Lee
- Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892
| | - Kyung Seok Kim
- School of Biological Sciences and Technology Chonnam National University, Gwangju 500-757, Korea
| | - Grzegorz Piszczek
- Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892
| | - Hyung Wook Nam
- Department of Biochemistry, College of Science, Protein Network Research Center, Yonsei University, Seoul 120-749, Korea
| | - Yu Sam Kim
- Department of Biochemistry, College of Science, Protein Network Research Center, Yonsei University, Seoul 120-749, Korea
| | - Taeho Ahn
- Department of Biochemistry, Chonnam National University, Gwangju 500-757, Korea
| | - Chul-Ho Yun
- School of Biological Sciences and Technology Chonnam National University, Gwangju 500-757, Korea
| | - Kanghwa Kim
- Department of Food and Nutrition and College of Veterinary Medicine Chonnam National University, Gwangju 500-757, Korea
| | - P Boon Chock
- Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892
| | - Ho Zoon Chae
- School of Biological Sciences and Technology Chonnam National University, Gwangju 500-757, Korea.
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Manandhar G, Miranda-Vizuete A, Pedrajas JR, Krause WJ, Zimmerman S, Sutovsky M, Sutovsky P. Peroxiredoxin 2 and peroxidase enzymatic activity of mammalian spermatozoa. Biol Reprod 2009; 80:1168-77. [PMID: 19208552 DOI: 10.1095/biolreprod.108.071738] [Citation(s) in RCA: 34] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/01/2022] Open
Abstract
Peroxiredoxin 2 (PRDX2) is a highly efficient redox protein that neutralizes hydrogen peroxide, resulting in protection of cells from oxidative damage and in regulation of peroxide-mediated signal transduction events. The oxidized form of PRDX2 is reverted back to the reduced form by the thioredoxin system. In the present study, we investigated the presence of PRDX2 in mouse and boar spermatozoa and in mouse spermatids using proteomic techniques and immunocytochemistry. Sperm and spermatid extracts displayed a 20-kDa PRDX2 band on Western blotting. PRDX2 occurred as a Triton-soluble form in spermatids and as a Triton-insoluble form in mature spermatozoa. Boar seminiferous tubule extracts were immunoprecipitated with PRDX2 antibody and separated by SDS-PAGE. Peptide mass fingerprinting by matrix-assisted laser desorption ionization-time of flight (TOF) and microsequencing by nanospray quadrupole-quadrupole TOF tandem mass spectrometry revealed the presence of PRDX2 ions in the immunoprecipitated band, along with sperm mitochondria-associated cysteine-rich protein, cellular nucleic acid-binding protein, and glutathione peroxidase 4. In mouse spermatocytes and spermatids, diffuse labeling of PRDX2 was observed in the cytoplasm and residual bodies. After spermiation, PRDX2 localization became confined to the mitochondrial sheath of the sperm tail midpiece. Boar spermatozoa displayed similar PRDX2 localization as in mouse spermatozoa. Boar spermatozoa with disrupted acrosomes expressed PRDX2 in the postacrosomal sheath region. Peroxidase enzyme activity of boar sperm extracts was evaluated by estimating the rate of NADPH oxidation in the presence or absence of a glutathione depletor (diethyl maleate) or a glutathione reductase inhibitor (carmustine). Diethyl maleate partially inhibited peroxidase activity, whereas carmustine showed an insignificant effect. These observations suggest that glutathione and glutathione reductase activity contribute only partially to the total peroxidase activity of the sperm extract. While the specific role of PRDX2 in the total peroxidase activity of sperm extract is still an open question, the present study for the first time (to our knowledge) shows the presence of PRDX2 in mammalian spermatozoa. Peroxidase activity in sperm extracts is not due to the glutathione system and therefore possibly involves PRDX2 and other peroxiredoxins.
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Affiliation(s)
- Gaurishankar Manandhar
- Division of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri 65211, USA.
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Eleuterio E, Di Giuseppe F, Sulpizio M, di Giacomo V, Rapino M, Cataldi A, Di Ilio C, Angelucci S. Proteome analysis of X-ray irradiated human erythroleukemia cells. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS 2008; 1784:611-20. [DOI: 10.1016/j.bbapap.2008.01.018] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 08/29/2007] [Revised: 12/28/2007] [Accepted: 01/20/2008] [Indexed: 11/16/2022]
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Zhang B, Wang Y, Liu K, Yang X, Song M, Wang Y, Bai Y. Adenovirus-mediated transfer of siRNA against peroxiredoxin I enhances the radiosensitivity of human intestinal cancer. Biochem Pharmacol 2008; 75:660-7. [DOI: 10.1016/j.bcp.2007.09.016] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/26/2007] [Revised: 09/10/2007] [Accepted: 09/17/2007] [Indexed: 10/22/2022]
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Bystrova MF, Budanova EN. Hydrogen peroxide and peroxiredoxins in redox regulation of intracellular signaling. BIOCHEMISTRY (MOSCOW) SUPPLEMENT SERIES A: MEMBRANE AND CELL BIOLOGY 2007. [DOI: 10.1134/s1990747807020018] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
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Lee W, Choi KS, Riddell J, Ip C, Ghosh D, Park JH, Park YM. Human peroxiredoxin 1 and 2 are not duplicate proteins: the unique presence of CYS83 in Prx1 underscores the structural and functional differences between Prx1 and Prx2. J Biol Chem 2007; 282:22011-22. [PMID: 17519234 DOI: 10.1074/jbc.m610330200] [Citation(s) in RCA: 120] [Impact Index Per Article: 6.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/11/2023] Open
Abstract
Human peroxiredoxins 1 and 2, also known as Prx1 and Prx2, are more than 90% homologous in their amino acid sequences. Prx1 and Prx2 are elevated in various cancers and are shown to influence diverse cellular processes. Although their growth regulatory role has traditionally been attributed to the peroxidase activity, the physiological significance of this function is unclear because the proteins are highly susceptible to inactivation by H(2)O(2). A chaperone activity appears to emerge when their peroxidase activity is lost. Structural studies suggest that they may form a homodimer or doughnut-shaped homodecamer. However, little information is available whether human Prx1 and Prx2 are duplicative in structure and function. We noted that Prx1 contains a cysteine (Cys(83)) at the putative dimer-dimer interface, which is absent in Prx2. We studied the role of Cys(83) in regulating the peroxidase and chaperone activities of Prx1, because the redox status of Cys(83) might influence the oligomeric structure and consequently the functions of Prx1. We show that Prx1 is more efficient as a molecular chaperone, whereas Prx2 is better suited as a peroxidase enzyme. Substituting Cys(83) with Ser(83) (Prx1C83S) results in dramatic changes in the structural and functional characteristics of Prx1 in a direction similar to those of Prx2. Here we also report the first crystal structure of human Prx1 and the presence of the Cys(83)-Cys(83) bond at the dimer-dimer interface of decameric Prx1. These findings are consistent with the hypothesis that human Prx1 and Prx2 possess unique functions and regulatory mechanisms and that Cys(83) bestows a distinctive identity to Prx1.
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Affiliation(s)
- Weonsup Lee
- Department of Cell Stress Biology, Roswell Park Cancer Institute, Buffalo, NY 14263, USA
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Abstract
Reactive oxygen species have been implicated in gametogenesis and embryo development in animals. As peroxiredoxins are now recognized as important protective antioxidant enzymes as well as modulators of hydrogen peroxide-mediated signaling, we addressed here the putative role of this novel family of peroxidases in gamete maturation and during embryogenesis in mammals and insects.
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Affiliation(s)
- Isabelle Donnay
- Veterinary Unit, Institut des Sciences de la Vie, Université catholique de Louvain, Louvain-La-Neuve, Belgium
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Abstract
We and others have shown that foam cell formation initiated by exposing macrophages to oxidized low density lipoprotein (oxLDL) triggers the differential expression of a number of proteins. Specifically, our experiments have identified peroxiredoxin I (Prx I) as one of these up-regulated proteins. The peroxiredoxins, a family of peroxidases initially described for their antioxidant capability, have generated recent interest for their potential to regulate signaling pathways. Those studies, however, have not examined peroxiredoxin for a potential dual functionality as both cytoprotective antioxidant and signal modulator in a single, oxidant-stressed system. In this report, we examine the up-regulation of Prx I in macrophages in response to oxLDL exposure and its ability to function as both antioxidant enzyme and regulator of p38 MAPK activation. As an antioxidant, induction of Prx I expression led to improved cell survival following treatment with oxLDL or tert-butyl hydroperoxide. The improved survival coincided with a decrease in measurable reactive oxygen species (ROS), and both the increased survival and reduced ROS were reversed by Prx I small interfering RNA transfection. Additionally, our data show that activation of p38 MAPK in oxLDL-treated macrophages was dependent on the up-regulation of Prx I. Reduction of Prx I expression by small interfering RNA transfection resulted in a significant decrease in p38 MAPK activation, whereas the up-regulation of Prx I expression with either oxLDL or ethoxyquin led to increased p38 MAPK activation. These results are consistent with multiple roles for Prx I in macrophage-derived foam cells that include functionality as both an antioxidant and a regulator of oxidant-sensitive signal transduction.
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Affiliation(s)
- James P Conway
- Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio 44106, USA
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Di Pietro R, Fang H, Fields K, Miller S, Flora M, Petricoin EC, Dveksler G, Rana RA, Grimley PM. Peroxiredoxin genes are not induced in myeloid leukemia cells exposed to ionizing radiation. Int J Immunopathol Pharmacol 2006; 19:517-24. [PMID: 17026836 DOI: 10.1177/039463200601900307] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023] Open
Abstract
Peroxiredoxins (Prx) comprise an extended family of small antioxidant proteins which conserve a thioredoxin-dependent catalytic function that can contribute to cell protection from reactive oxygen species (ROS). ROS generation is one of the deleterious intracellular effects of ionizing radiation, but the role of Prx during radiation treatment has not been extensively explored. Present experiments measure effects of ionizing radiation on expression of human Prx types I (PAGA), II (NKEF-B) and IV (AOE372) in human myeloid leukemia cells (K562). Prx gene transcription was analyzed by amplifying with RT-PCR cDNAs complementary to each Prx-specific coding sequence and by identifying the derived products with Southern blotting procedure. Transcripts of GAPDH were used as the endogenous standard for semi-quantitative comparisons. No consistent increase in Prx gene expression was detected at time intervals up to 72 h after gamma radiation doses that caused cell cycle arrest and nuclear damage (maximum 20 Gy). Immunoblots also were consistent with a prolonged expression or stability of the Prx I/II proteins. Similarly, a cytotoxic concentration of the oxidant hemin, which stimulates rapid hemoglobinization of K562 cells, caused no induction of Prx gene expression. Our results indicate a high Prx stability in human radio-resistant leukemia cells.
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Affiliation(s)
- R Di Pietro
- Department of Biomorphology, G. d'Annunzio University, Chieti-Pescara, Italy.
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An JH, Seong JS. Proteomics analysis of apoptosis-regulating proteins in tissues with different radiosensitivity. JOURNAL OF RADIATION RESEARCH 2006; 47:147-55. [PMID: 16819141 DOI: 10.1269/jrr.47.147] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/10/2023]
Abstract
The aim of this study was to identify of radiosusceptibility proteins in tissues with different radiosensitivity. C3H/HeJ mice were exposed to 10 Gy. The tissues were processed for proteins extraction and were analyzed by 2-dimensional electrophoresis. The proteins were identified by matrix-assisted laser desorption ionizing time-of-flight mass spectrometry and validated by immunohistochemical staining and Western blotting. The peaks of apoptosis levels were 35.3 +/- 1.7% and 0.6 +/- 0.2% in the spleen and the liver, respectively, after ionizing radiation. Analysis of liver tissue showed that the expression level of ROS related proteins such as cytochrome c, glutathione S transferase, NADH dehydrogenase and peroxiredoxin VI increased after radiation. The expression level of cytochrome c increased to 3-fold after ionizing radiation in both tissues. However in spleen tissue, the expression level of various kinds of apoptosis regulating proteins increased after radiation. These involved iodothyronine, CD 59A glycoprotein precursor, fas antigen and tumor necrosis factor -inducible protein TSG-6n precursor after radiation. The difference in the apoptosis index between the liver and spleen tissues is closely associated with the expression of various kinds of apoptosis-related proteins. The result suggests that the expression of apoptosis-related protein and redox proteins play important roles in this radiosusceptibility.
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Affiliation(s)
- Jeung Hee An
- Department of Radiation Oncology, Brain Korea 21 Project for Medical Science, Yonsei University Medical College, Seoul Korea.
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Abstract
Oxidative damage is one threat spermatozoa have to face during epididymal maturation and storage. However, it is clear that reactive oxygen species (ROS) are also central for sperm physiology in processes such as sperm maturation and capacitation. It is therefore essential that there exists around sperm cells a fine balance between ROS production and recycling. To do so, sperm cells and epididymal epithelial cells rely on common enzymatic ROS scavengers such as superoxide dismutase (SOD), glutathione peroxidases (GPX) and catalase (CAT) as well as more specific types such as indoleamine dioxygenase (IDO). Among the catalytic triad (SOD/GPX/CAT), the glutathione peroxidase protein family occupies a peculiar position, since several GPX have been found to be present on and around epididymal transiting sperm cells. Here, we will review our present knowledge regarding GPX expression, presence and putative role(s) within the epididymis and on spermatozoa. Taking into account our recent findings regarding the epididymal expression of indoleamine dioxygenase in mouse we will also discuss how we think this superoxide anion recycling enzyme completes the complex ROS generation/recycling balance in this organ.
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Affiliation(s)
- Joël R Drevet
- Université Blaise Pascal, CNRS UMR 6547 GEEM, 24 Avenue des Landais, 63177 Aubière, France.
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47
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Zhang B, Su Y, Ai G, Wang Y, Wang T, Wang F. Involvement of peroxiredoxin I in protecting cells from radiation-induced death. JOURNAL OF RADIATION RESEARCH 2005; 46:305-12. [PMID: 16210786 DOI: 10.1269/jrr.46.305] [Citation(s) in RCA: 39] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/04/2023]
Abstract
Peroxiredoxin I (Prx-I), a key member of the peroxiredoxin family, reduces peroxides and equivalents through the thioredoxin system. Our previous work has shown that expression of Prx-I in mammalian cells increases following ionizing radiation (IR), indicating that Prx-I actively responds to IR-induced reactive oxygen species (ROS) and suggesting that Prx-I plays an important role in protecting cells from IR-induced death. To test this hypothesis, we suppressed the expression of Prx-I in SW480 cells by RNA interference. Our results show that IR induces the expression of Prx-I in SW480 cells in a dose- and time-dependent manner. The recombinant siRNA vector targeting Prx-I dramatically reduced the expression of Prx-I in SW480 cells. When Prx-I was knocked down in SW480 cells, the cells exhibited a decreased growth rate, a reduced antioxidant capability following IR and became more sensitive to IR-induced apoptosis. Together, our results demonstrate that Prx-I plays an important role in protecting cells from IR-induced cell death, which might be through scavenging IR-induced ROS in the cells.
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Affiliation(s)
- Bo Zhang
- Combined Injury Institute of Third Military Medical University, State Key Laboratory of Trauma, Burns and Combined Injury
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48
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Abstract
Since radiation treatment has been reappraised in the treatment of hepatic tumors, radiation response in the liver is emerging as an interesting new area of investigation. In this study, identification of the repertoire of signaling proteins was performed using a proteomics approach involving cellular responses of liver tissue to ionizing radiation. Approximately 800 protein spots were detected. Among them, at least 28 proteins showed significant quantitative alterations after radiation. The significantly altered proteins were categorized as those related to reactive oxygen species (ROS) metabolism, metabolic pathway proteins, and G-type proteins. Particularly, the expression levels of proteins related to ROS metabolism, including cytochrome c, glutathione S-transferase Pi, NADH dehydrogenase, and peroxiredoxin VI, were increased after radiation. It is suggested that although radiation initiates cytotoxic effects, it can also induce a radioprotective antioxidant system.
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Affiliation(s)
- Jeung Hee An
- Department of Radiation Oncology, Brain Korea 21 Project for Medical Science, Yonsei University Medical College, Shinchon-dong 134, Seodamun-Ku, Seoul 120-752, Korea
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49
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Abstract
Peroxiredoxins (Prxs) are a family of multifunctional antioxidant thioredoxin-dependent peroxidases that have been identified in a large variety of organisms. The major functions of Prxs comprise cellular protection against oxidative stress, modulation of intracellular signaling cascades that apply hydrogen peroxide as a second messenger molecule, and regulation of cell proliferation. In the present review, we discuss pertinent findings on the protein structure, the cell- and tissue-specific distribution, as well as the subcellular localization of Prxs. A particular emphasis is put on Prx I, which is the most abundant and ubiquitously distributed member of the mammalian Prxs. Major transcriptional and posttranslational regulatory mechanisms and signaling pathways that control Prx gene expression and activity are summarized. The interaction of Prx I with the oncogene products c-Abl and c-Myc and the regulatory role of Prx I for cell proliferation and apoptosis are highlighted. Recent findings on phenotypical alterations of mouse models with targeted disruptions of Prx genes are discussed, confirming the physiological functions of Prxs for antioxidant cell and tissue protection along with an important role as tumor suppressors.
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Affiliation(s)
- Stephan Immenschuh
- Institut für Klinische Immunologie und Transfusionsmedizin, Justus-Liebig-Universität Giessen, Giessen, Germany.
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50
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Zhang B, Su YP, Wang T, Wang FC, Ai GP, Xu H, Wang JP, Huang YS, Jiang JX. Cloning and expression of mouse peroxiredoxin I in IEC-6 Cells. World J Gastroenterol 2004; 10:2109-12. [PMID: 15237445 PMCID: PMC4572344 DOI: 10.3748/wjg.v10.i14.2109] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
AIM: To clone and express mouse peroxiredoxin I in IEC-6 cells.
METHODS: Total RNAs were isolated from cultured IEC-6 cells, and the coding region of peroxiredoxin I was amplified by RT-PCR. After it was cloned into T-vector and sequenced, pSG5 was used to transiently express peroxiredoxin I in IEC-6 by liposome-mediated transfection, and the expression of peroxiredoxin I was evaluated by RT-PCR and Western blot.
RESULTS: A DNA fragment about 750 bp was amplified from total RNAs of IEC-6 cells using specific primers of peroxiredoxin I. The sequencing confirmed the coding region was successfully cloned into T-vector, which was completely coincident with the sequence in GeneBank. After the EcoR I-BamH I fragment of T-vector containing peroxiredoxin I was inserted into pSG5, the recombinant plasmid was transferred to IEC-6 cells. RT-PCR assay showed that a DNA fragment of 930 bp could be amplified, which indicated the transcription of pSG5-Prx. Western blot confirmed the expression of peroxiredoxin I in IEC-6 cells.
CONCLUSION: Mouse peroxiredoxin I can be successfully expressed in IEC-6 cells.
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Affiliation(s)
- Bo Zhang
- Institute of Combined Injury of PLA, State Key Laboratory of Trauma, Third Military Medical University, Gaotanyan Street 30, Chongqing 400038, China
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