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Park J, An G, Lee H, Park S, Ham J, Bazer FW, Song G, Lim W. Beta-cyfluthrin impairs implantation process by inducing mitochondrial defects and changes in reactive oxygen species-mediated signaling pathways in porcine trophectoderm and uterine luminal epithelial cells. THE SCIENCE OF THE TOTAL ENVIRONMENT 2024; 934:173097. [PMID: 38729356 DOI: 10.1016/j.scitotenv.2024.173097] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/06/2024] [Revised: 04/22/2024] [Accepted: 05/07/2024] [Indexed: 05/12/2024]
Abstract
Pyrethroid insecticides, such as beta-cyfluthrin, are used extensively globally, including in households and agriculture, and have been detected in the milk and urine of humans and cattle. Beta-cyfluthrin exhibits toxic effects, including neurotoxicity and male reproductive toxicity; however, few studies have investigated female reproductive toxicity despite its wide environmental distribution. The present study investigates effects of beta-cyfluthrin on implantation in porcine cells (pTr from the trophectoderm and pLE from the endometrial luminal epithelium). To identify the various physiological changes induced by beta-cyfluthrin, such as apoptosis and lipid peroxidation, flow cytometry analysis and immunofluorescence were performed with various reagents. In addition, the expression of genes and proteins associated with intracellular changes was confirmed using qRT-PCR and western blotting. Beta-cyfluthrin induced cell-cycle arrest and altered intracellular calcium flux. It also disrupted the mitochondrial function and promoted reactive oxygen species (ROS) production, leading to lipid peroxidation. Moreover, ROS induced by beta-cyfluthrin altered mitogen-activated protein kinase (MAPK) pathways and decreased cell migration capability. The expression levels of genes that are significant during early pregnancy were altered by beta-cyfluthrin in both cell lines. The changes resulted in apoptosis and diminished cell proliferation of pTr and pLE. Collectively, the results imply that beta-cyfluthrin disrupts the implantation process by affecting the physiology of the trophectoderm and endometrial luminal epithelial cells. The present study is the first to reveal the cellular mechanisms of beta-cyfluthrin on the female reproductive system and highlights the need for further in-depth research into its hazards.
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Affiliation(s)
- Junho Park
- Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea
| | - Garam An
- Department of Biological Sciences, Institute of Basic Science, Sungkyunkwan University, Suwon 16419, Republic of Korea
| | - Hojun Lee
- Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea
| | - Sunwoo Park
- Department of Plant & Biomaterials Science, Gyeongsang National University, Jinju-si, Gyeongnam 52725, Republic of Korea
| | - Jiyeon Ham
- Division of Animal and Dairy Science, Chungnam National University, Daejeon 34134, Republic of Korea
| | - Fuller W Bazer
- Department of Animal Science, Texas A&M University, College Station, TX 77843-2471, USA
| | - Gwonhwa Song
- Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea.
| | - Whasun Lim
- Department of Biological Sciences, Institute of Basic Science, Sungkyunkwan University, Suwon 16419, Republic of Korea.
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Lestari B, Fukushima T, Utomo RY, Wahyuningsih MSH. Apoptotic and non-apoptotic roles of caspases in placenta physiology and pathology. Placenta 2024; 151:37-47. [PMID: 38703713 DOI: 10.1016/j.placenta.2024.03.013] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/15/2023] [Revised: 03/12/2024] [Accepted: 03/27/2024] [Indexed: 05/06/2024]
Abstract
Caspases, a family of cysteine proteases, are pivotal regulators of apoptosis, the tightly controlled cell death process crucial for eliminating excessive or unnecessary cells during development, including placental development. Collecting research has unveiled the multifaceted roles of caspases in the placenta, extending beyond apoptosis. Apart from their involvement in placental tissue remodeling via apoptosis, caspases actively participate in essential regulatory processes, such as trophoblast fusion and differentiation, significantly influencing placental growth and functionality. In addition, growing evidence indicates an elevation in caspase activity under pathological conditions like pre-eclampsia (PE) and intrauterine growth restriction (IUGR), leading to excessive cell death as well as inflammation. Drawing from advancements in caspase research and placental development under both normal and abnormal conditions, we examine the significance of caspases in both cell death (apoptosis) and non-cell death-related processes within the placenta. We also discuss potential therapeutics targeting caspase-related pathways for placenta disorders.
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Affiliation(s)
- Beni Lestari
- Department Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia; Cancer Chemoprevention Research Center, Universitas Gadjah Mada, Yogyakarta, Indonesia
| | - Toshiaki Fukushima
- Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, Japan.
| | - Rohmad Yudi Utomo
- Cancer Chemoprevention Research Center, Universitas Gadjah Mada, Yogyakarta, Indonesia; Department Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia
| | - Mae Sri Hartati Wahyuningsih
- Department Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.
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Gajić M, Schröder-Heurich B, Mayer-Pickel K. Deciphering the immunological interactions: targeting preeclampsia with Hydroxychloroquine's biological mechanisms. Front Pharmacol 2024; 15:1298928. [PMID: 38375029 PMCID: PMC10875033 DOI: 10.3389/fphar.2024.1298928] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/22/2023] [Accepted: 01/23/2024] [Indexed: 02/21/2024] Open
Abstract
Preeclampsia (PE) is a complex pregnancy-related disorder characterized by hypertension, followed by organ dysfunction and uteroplacental abnormalities. It remains a major cause of maternal and neonatal morbidity and mortality worldwide. Although the pathophysiology of PE has not been fully elucidated, a two-stage model has been proposed. In this model, a poorly perfused placenta releases various factors into the maternal circulation during the first stage, including pro-inflammatory cytokines, anti-angiogenic factors, and damage-associated molecular patterns into the maternal circulation. In the second stage, these factors lead to a systemic vascular dysfunction with consecutive clinical maternal and/or fetal manifestations. Despite advances in feto-maternal management, effective prophylactic and therapeutic options for PE are still lacking. Since termination of pregnancy is the only curative therapy, regardless of gestational age, new treatment/prophylactic options are urgently needed. Hydroxychloroquine (HCQ) is mainly used to treat malaria as well as certain autoimmune conditions such as systemic lupus and rheumatoid arthritis. The exact mechanism of action of HCQ is not fully understood, but several mechanisms of action have been proposed based on its pharmacological properties. Interestingly, many of them might counteract the proposed processes involved in the development of PE. Therefore, based on a literature review, we aimed to investigate the interrelated biological processes of HCQ and PE and to identify potential molecular targets in these processes.
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Affiliation(s)
- Maja Gajić
- Department of Obstetrics and Gynecology, Medical University of Graz, Graz, Austria
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He B, Bai J, Wu Z. Glucosamine enhances proliferation, barrier, and anti-oxidative functions in porcine trophectoderm cells. Food Funct 2022; 13:4551-4561. [PMID: 35352734 DOI: 10.1039/d1fo04086c] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
Trophectoderm (TE) is the first epithelium that appears during mammalian embryogenesis, and is a polarized transporting single cell layer that comprises the wall of the blastocyst. Previous studies have revealed the functional roles of glucose (Gluc), fructose (Fruc), and glutamine (Gln), which play a positive role in porcine trophectoderm (pTr) cell proliferation and migration, suggesting the importance of nutrients for normal development of the conceptus and implantation. This work was conducted to test the hypothesis that glucosamine (GlcN), which is synthesized from Gln and Fruc-6-phosphate through the hexosamine biosynthesis pathway (HBP), can stimulate proliferation and sustain the barrier and anti-oxidative functions of pTr cells. Cells were treated with 0, 0.25, or 0.5 mmol L-1 GlcN in the presence or absence of adiquat (DQ) for the indicated time points. The results showed that 0.25 or 0.5 mmol L-1 GlcN stimulated pTr cell viability and DNA replication compared to the control group. The addition of 0.25 mmol L-1 GlcN enhanced the phosphorylation of mTOR signaling proteins, which can be inhibited by the inhibitor of phosphatidylinositol 3-kinase (PI3K), LY294002. Transepithelial electrical resistance (TEER) was increased, and paracellular permeability was correspondingly reduced in GlcN treatment. GlcN attenuated DQ-induced cell death and reduced the level of reactive oxygen species (ROS). The decreased TEER values and increased paracellular permeability caused by DQ treatment were also inhibited by GlcN treatment. The addition of 0.5 mmol L-1 GlcN increased the protein expression of zonula occludens-3 (ZO-3), claudin-3, and claudin-4 in pTr cells, while inhibited the downregulation protein of claudin-1 and claudin-3 brought about by oxidative stress. Collectively, GlcN plays an important role in promoting proliferation and stimulating the mTOR cell signaling pathway, as well as ameliorating oxidative stress and augmenting barrier functions in pTr cells.
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Affiliation(s)
- Beibei He
- State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, China 100193.
| | - Jun Bai
- State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, China 100193.
| | - Zhenlong Wu
- State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, China 100193.
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Redman CW, Staff AC, Roberts JM. Syncytiotrophoblast stress in preeclampsia: the convergence point for multiple pathways. Am J Obstet Gynecol 2022; 226:S907-S927. [PMID: 33546842 DOI: 10.1016/j.ajog.2020.09.047] [Citation(s) in RCA: 186] [Impact Index Per Article: 62.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/05/2020] [Revised: 09/14/2020] [Accepted: 09/19/2020] [Indexed: 12/29/2022]
Abstract
Preeclampsia evolves in 2 stages: a placental problem that generates signals to the mother to cause a range of responses that comprise the second stage (preeclampsia syndrome). The first stage of early-onset preeclampsia is poor placentation, which we here call malplacentation. The spiral arteries are incompletely remodeled, leading to later placental malperfusion, relatively early in the second half of pregnancy. The long duration of the first stage (several months) is unsurprisingly associated with fetal growth restriction. The first stage of late-onset preeclampsia, approximately 80% of total cases, is shorter (several weeks) and part of a process that is common to all pregnancies. Placental function declines as it outgrows uterine capacity, with increasing chorionic villous packing, compression of the intervillous space, and fetal hypoxia, and causes late-onset clinical presentations such as "unexplained" stillbirths, late-onset fetal growth restriction, or preeclampsia. The second stages of early- and late-onset preeclampsia share syncytiotrophoblast stress as the most relevant feature that causes the maternal syndrome. Syncytiotrophoblast stress signals in the maternal circulation are probably the most specific biomarkers for preeclampsia. In addition, soluble fms-like tyrosine kinase-1 (mainly produced by syncytiotrophoblast) is the best-known biomarker and is routinely used in clinical practice in many locations. How the stress signals change over time in normal pregnancies indicates that syncytiotrophoblast stress begins on average at 30 to 32 weeks' gestation and progresses to term. At term, syncytiotrophoblast shows increasing markers of stress, including apoptosis, pyroptosis, autophagy, syncytial knots, and necrosis. We label this phenotype the "twilight placenta" and argue that it accounts for the clinical problems of postmature pregnancies. Senescence as a stress response differs in multinuclear syncytiotrophoblast from that of mononuclear cells. Syncytiotrophoblast irreversibly acquires part of the senescence phenotype (cell cycle arrest) when it is formed by cell fusion. The 2 pathways converge on the common pathologic endpoint, syncytiotrophoblast stress, and contribute to preeclampsia subtypes. We highlight that the well-known heterogeneity of the preeclampsia syndrome arises from different pathways to this common endpoint, influenced by maternal genetics, epigenetics, lifestyle, and environmental factors with different fetal and maternal responses to the ensuing insults. This complexity mandates a reassessment of our approach to predicting and preventing preeclampsia, and we summarize research priorities to maximize what we can learn about these important issues.
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Pathare-Ingawale P, Chavan-Gautam P. The balance between cell survival and death in the placenta: Do neurotrophins have a role? Syst Biol Reprod Med 2021; 68:3-12. [PMID: 34615417 DOI: 10.1080/19396368.2021.1980132] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/08/2022]
Abstract
Neurotrophins (NT) are a closely related family of growth factors, which regulate the nervous system's development, maintenance, and function. Although NTs have been well studied in neuronal cells, they are also expressed in the placenta. Despite their suggested role in regulating fetoplacental development, their precise functional significance in the placenta remains elusive. NT activate two different classes of receptors. These include the Trk, tropomyosin-related kinase family of high-affinity tropomyosin-related kinase receptors, which induces cell survival, and the p75NTR, p75 neurotrophin receptor, a member of the tumor necrosis factor(TNF) receptor superfamily, which induces apoptosis in neuronal cells. Mature NT molecule results from proteolysis of a biologically active precursor form called pro-neurotrophins (pro-NT) by the intracellular proprotein convertase or furin. Pro-NTs have a regulatory role in determining cell survival and apoptosis. Here, we review the literature on the expression and functions of NTs and their receptors in the placenta and discuss their possible role in placental tissue development and apoptosis. The possible implications of imbalance in pro-NT and mature-NT levels for fetoplacental development are also discussed.Abbreviations AGE/ALEs: Advanced glycation/lipoxidation end products; Bax: Bcl 2 Associated X; Bcl-2: B-cell lymphoma 2; BDNF: Brain-derived neurotrophic factor; FAS/FASL: Fas cell surface death receptor/ ligand; IUGR: Intrauterine growth restriction; JNK: c-Jun amino-terminal kinase; MAP: mitogen-activated protein k; mRNA: Messenger ribonucleic acid; NGF: Nerve growth factor; NT: Neurotrophins; NRAGE: Neurotrophin receptor-interacting MAGE homolog; NRIF: Neurotrophin receptor interacting factor; PE: Preeclampsia; PI3k: Phosphoinositide 3- kinase; PLC: Phospholipase C; p75NTR: p75 neurotrophin receptor; Pro-NT: Pro-neurotrophins; PTB: Preterm birth; p53: Tumor protein p53; TNF: Tumor necrosis factor; TRAF: TNFR-associated factors; Trk: Tropomyosin-related kinase; siRNA: small interfering ribonucleic acid.
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Affiliation(s)
| | - Preeti Chavan-Gautam
- Interdisciplinary School of Health Science, Savitribai Phule Pune University, Pune, India
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Kovács P, Joó JG, Tamás V, Molnár Z, Burik-Hajas D, Bódis J, Kornya L. The role of apoptosis in the complex pathogenesis of the most common obstetrics and gynaecology diseases. Physiol Int 2021; 107:106-119. [PMID: 32491289 DOI: 10.1556/2060.2020.00014] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/09/2019] [Accepted: 01/07/2020] [Indexed: 11/19/2022]
Abstract
Purpose We aimed to assess the etiological role of apoptotic genes Bcl-2 and Bax in the background of major obstetric and gynaecological diseases. Methods Placental tissue samples were collected from 101 pregnancies with intrauterine growth restriction and 104 pregnancies with premature birth with 140 controll samples from term, eutrophic newborns. In addition, gene expression assessment of the genes Bax and Bcl-2 was performed in 101 uterine leiomyoma tissue samples at our disposal with 110 control cases. Gene expression levels were assessed by PCR method. Results The expression of the Bcl-2 gene was decreased in placental samples with intrauterine growth restriction. Significant overexpression of the proapoptotic Bax gene was detected in samples from premature infants. Antiapoptotic Bcl-2 gene expression was found to be significantly increased in fibroid tissues. Conclusion Apoptosis plays a crucial role in the development of the most common OB/GYN conditions. Decrease in the placental expression of the antiapoptotic gene Bcl-2 may upset the balance of programmed cell death.
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Affiliation(s)
- P Kovács
- 1Clinical Research Units Hungary, Miskolc, Hungary.,2Doctoral School of Health Sciences, Faculty of Health Sciences, University of Pécs, Pécs, Hungary
| | - József Gábor Joó
- 2Doctoral School of Health Sciences, Faculty of Health Sciences, University of Pécs, Pécs, Hungary.,3First Department of Obstetrics and Gynaecology, Semmelweis University, Budapest, Hungary
| | - V Tamás
- 2Doctoral School of Health Sciences, Faculty of Health Sciences, University of Pécs, Pécs, Hungary
| | - Z Molnár
- 2Doctoral School of Health Sciences, Faculty of Health Sciences, University of Pécs, Pécs, Hungary
| | - D Burik-Hajas
- 2Doctoral School of Health Sciences, Faculty of Health Sciences, University of Pécs, Pécs, Hungary
| | - J Bódis
- 2Doctoral School of Health Sciences, Faculty of Health Sciences, University of Pécs, Pécs, Hungary.,4Hungarian Academy of Sciences, University of Pécs (MTA-PTE), Human Reproduction Scientific Research Group, University of Pécs, Pécs, Hungary
| | - L Kornya
- 2Doctoral School of Health Sciences, Faculty of Health Sciences, University of Pécs, Pécs, Hungary.,5Central Hospital of Southern Pest, National Institute of Hematology and Infectious Diseases, Budapest, Hungary
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Anti-Apoptotic Effect of Apelin in Human Placenta: Studies on BeWo Cells and Villous Explants from Third-Trimester Human Pregnancy. Int J Mol Sci 2021; 22:ijms22052760. [PMID: 33803239 PMCID: PMC7967155 DOI: 10.3390/ijms22052760] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/16/2021] [Revised: 03/01/2021] [Accepted: 03/03/2021] [Indexed: 12/19/2022] Open
Abstract
Previously, we demonstrated the expression of apelin and G-protein-coupled receptor APJ in human placenta cell lines as well as its direct action on placenta cell proliferation and endocrinology. The objective of this study was to examine the effect of apelin on placenta apoptosis in BeWo cells and villous explants from the human third trimester of pregnancy. The BeWo cells and villous explants were incubated with apelin (2 and 20 ng/mL) alone or with staurosporine for 24 to 72 h. First, we analysed the dose- and time-dependent effect of apelin on the expression of apoptotic factors on the mRNA level by real-time PCR and on the protein level using Western blot. Next, we checked caspase 3 and 7 activity by Caspase-Glo 3/7, DNA fragmentation by the Cell Death Detection ELISA kit and oxygen consumption by the MitoXpress-Xtra Oxygen Consumption assay. We found that apelin increased the expression of pro-survival and decreased proapoptotic factors on mRNA and protein levels in both BeWo cells and villous explants. Additionally, apelin inhibited caspase 3 and 7 activity and DNA fragmentation in staurosporine-induced apoptosis as also attenuated oxidative stress by increasing extracellular oxygen consumption. The antiapoptotic effect of apelin in BeWo cells was mediated by the APJ receptor and mitogen-activated protein kinase (ERK1/2/MAP3/1) and protein kinase B (AKT). The obtained results showed the antiapoptotic effect of apelin on trophoblast cells, suggesting its participation in the development of the placenta.
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Elsalam SA, Mansor AE, Sarhan MH, Shalaby AM, Gobran MA, Alabiad MA. Evaluation of Apoptosis, Proliferation, and Adhesion Molecule Expression in Trophoblastic Tissue of Women With Recurrent Spontaneous Abortion and Infected With Toxoplasma gondii. Int J Gynecol Pathol 2021; 40:124-133. [PMID: 32833877 DOI: 10.1097/pgp.0000000000000683] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2022]
Abstract
Recurrent spontaneous abortion is an obstetric complication with undefined causes. Apoptosis, proliferation, and adhesion are considered important factors in the pathogenesis of abortion. This work aimed to determine Bax and Bcl-2 as a proapoptotic and antiapoptotic protein, Ki67 and P27kip as proliferative and antiproliferative proteins, and E-cadherin and CD44 as adhesion molecules in the trophoblastic tissues in cases with recurrent miscarriage. Immunohistochemistry and quantitative polymerase chain reaction analysis of Bax, Bcl-2, Ki67, P27kip, E-cadherin, and CD44 in paraffin-embedded sections of placental tissues obtained from 108 women were divided into 3 categories: 66 Toxoplasma gondii-positive women with recurrent abortion, 22 T. gondii-negative women with recurrent abortion, and 20 women with no history of abortion as a control group. The mean ratio of the expression of Bax and P27kip proteins was 35.3% and 36.1%, which is significantly higher than that of the second group (19.88 and 20.02%), and the third group (12.3% and 10.98%), while the mean ratio of the expression of Bcl-2, Ki67, E-cadherin, and CD44 proteins was 12.35%, 11.23%, 10.32%, and 9.97%, which is significantly lower than that of the second group (33.75%, 13.18%, 21.88%, and 23.29%) and that of the third group (38.58%, 39.27%, 37.98%, and 35.79%). The presence of proapoptotic protein (Bax) and antiproliferative protein (P27kip) at high levels and the presence of antiapoptotic protein (Bcl-2), proliferative protein (Ki67), and adhesion molecules (E-cadherin and CD44) in lower levels in the T. gondii-positive group clarify the mechanism involved in the induction of abortion and loss of pregnancy.
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Liu T, Ma Y, Yin Q, Zhou H, Fang Y. Association of β-arrestin1 and p53-Mdm2 signaling in the development of missed abortion. J Obstet Gynaecol Res 2021; 47:1675-1685. [PMID: 33611816 DOI: 10.1111/jog.14643] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2020] [Revised: 12/02/2020] [Accepted: 12/17/2020] [Indexed: 12/14/2022]
Abstract
BACKGROUND Missed abortion is a peculiar form of spontaneous abortion before 20 weeks' gestation. The definite etiology and pathogenesis are not fully understood. Recent studies have demonstrated that p53/Mdm2-mediated ubiquitination of the IGF-1R may be closely related to G-protein-coupled receptor kinases (GRK)/β-arrestin1 system. Our previous studies have confirmed that the elevated expression of p53 and Mdm2 may be responsible for apoptosis during missed abortion. However, there was no information surrounding β-arrestin1 in missed abortion. METHODS The mRNA levels of β-arrestin1 in villous samples of 30 missed abortion patients and 31 healthy controls were determined by real-time quantitative polymerase chain reaction (PCR). Immunohistochemistry was used to explore the expression and location of β-arrestin1, p53, Mdm2, VEGF and HIF-lα in trophoblasts. Transwell assays were performed to examine the influences of β-arrestin1 expression on cell invasion. Furthermore, we tested the effect of β-arrestin1 on the expression of p53, Mdm2, ERK, AKT and NF-κB. RESULTS The expression of β-arrestin1 in the villous samples of missed abortion group was dramatically lower than control group by quantitative real-time-PCR and immunohistochemistry. Furthermore, the patients with missed abortion showed significantly higher levels of p53, Mdm2, HIF-lα and lower level of VEGF than healthy controls by immunohistochemistry. Functional studies showed that suppression of β-arrestin1 in HTR-8 cells inhibited cell invasion. The protein expressions of ERK and AKT in HTR-8 cells were significantly downregulated by reducing the expression of β-arrestin1, while the expressions of p53, Mdm2, NF-κB were enhanced. Overexpression of β-arrestin1 exhibited the adverse effect. CONCLUSION Our data indicated that β-arrestin1 play an important role in maintaining the maternal-fetal tolerance, the decreased expression of β-arrestin1 in the villous samples may be related with the development of missed abortion.
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Affiliation(s)
- Ting Liu
- Department of Gynecology and Obstetrics, Qilu Hospital of Shandong University, Jinan, Shandong, China
| | - Yuyan Ma
- Department of Gynecology and Obstetrics, Qilu Hospital of Shandong University, Jinan, Shandong, China
| | - Qihui Yin
- Department of Gynecology and Obstetrics, Qilu Hospital of Shandong University, Jinan, Shandong, China
| | - Huanyu Zhou
- Department of Gynecology and Obstetrics, Qilu Hospital of Shandong University, Jinan, Shandong, China
| | - Yan Fang
- Department of Gynecology and Obstetrics, Qilu Hospital of Shandong University, Jinan, Shandong, China
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Wang R, Huang X, Ma C, Zhang H. Toxicological Effects of BPDE on Dysfunctions of Female Trophoblast Cells. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2021; 1300:151-160. [PMID: 33523433 DOI: 10.1007/978-981-33-4187-6_7] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Abstract
Polycyclic aromatic hydrocarbons (PAHs) are widely spread persistent environmental toxicants. Its typical representative benzo[a]pyrene (BaP) is a human carcinogen. BaP can pass through the placental barrier and is finally metabolized into benzo[a]pyren-7, 8-dihydrodiol-9, 10-epoxide (BPDE). BPDE can form DNA adducts, which directly affect the female reproductive health. Based on the special physiological functions of trophoblast cells and its important effect on normal pregnancy, this chapter describes the toxicity and molecular mechanism of BPDE-induced dysfunctions of trophoblast cells. By affecting the invasion, migration, apoptosis, proliferation, inflammation, and hormone secretion of trophoblast cells, BPDE causes diseases such as choriocarcinoma, intrauterine growth restriction, eclampsia, and abortion. In the end, it is expected to provide a scientific basis and prevention approach for women's reproductive health and decision-making basis for the formulation of environmental health standards.
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Affiliation(s)
- Rong Wang
- Key Laboratory of Environment and Female Reproductive Health, West China School of Public Health, Sichuan University, Chengdu, China
| | - Xinying Huang
- Key Laboratory of Environment and Female Reproductive Health, West China School of Public Health, Sichuan University, Chengdu, China
| | - Chenglong Ma
- Key Laboratory of Environment and Female Reproductive Health, West China School of Public Health, Sichuan University, Chengdu, China
| | - Huidong Zhang
- Key Laboratory of Environment and Female Reproductive Health, West China School of Public Health, Sichuan University, Chengdu, China.
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Fang J, Xie B, Chen B, Qiao C, Zheng B, Luan X, Liu J, Yan Y, Zheng Q, Wang M, Chen W, He Z, Shen C, Li H, Chen X, Yu J. Biochemical clinical factors associated with missed abortion independent of maternal age: A retrospective study of 795 cases with missed abortion and 694 cases with normal pregnancy. Medicine (Baltimore) 2018; 97:e13573. [PMID: 30558023 PMCID: PMC6320114 DOI: 10.1097/md.0000000000013573] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/25/2022] Open
Abstract
The incidence of fertile women with missed abortion dramatically increased in recent years, while very few serum indices have been identified for the diagnosis of missed abortion. The aim of this study was to identify related factors for missed abortion through a retrospective study of serum indices.A total of 795 cases of women with missed abortion and 694 cases of women with normal pregnancy between March 2014 and March 2017 were included in the present study. The diagnosis of missed abortion was based on clinical history, clinical examination, and transvaginal ultrasound findings. The final diagnosis of missed abortion was based on assessment of pregnancy structures (i.e., a gestational sac without fetal heart rate) via transvaginal ultrasound. We evaluated the clinical values of 4 serum indices and their relationship to missed abortion: gamma-glutamyltransferase (GGT), lactate dehydrogenase (LDH), adenosine deaminase (ADA), and fibrinogen (FIB).The serum levels of GGT, ADA, and FIB showed statistically significant differences comparing women who experienced missed abortion with women who had normal pregnancies (controls). Among women with missed abortion, the levels of GGT and ADA were dramatically increased (GGT: P < .0001; ADA: P = .0459), while FIB levels were slightly lower (P = .0084) compared to controls. The LDH levels exhibited a non-significant trend toward lower levels in the missed abortion group (P = .3951). Interestingly, the observed significant increase in serum GTT levels among women with missed abortion was not affected by maternal age.This study found that GTT may be a useful marker which was associated with missed abortion, indicating its potential clinical roles in missed abortion.
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Affiliation(s)
- Jie Fang
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
| | - Bing Xie
- Department of Obstetrics and Gynecology, the Fourth People's Hospital of Zhenjiang, Zhenjiang
| | - Binghai Chen
- Department of Urology, Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
| | - Chen Qiao
- Department of Clinical Pharmacy, Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
| | - Bo Zheng
- Center for Reproduction and Genetics, Suzhou Municipal Hospital, the Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou
| | - Xiaojin Luan
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
| | - Jiajia Liu
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
| | - Yidan Yan
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
| | - Qianwen Zheng
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
- Reproductive Sciences Institute of Jiangsu University, Jiangsu University, Zhenjiang, China
| | - Min Wang
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
| | - Wanyin Chen
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
| | - Zeyu He
- Department of Clinical Medicine, China Medical University, Shenyang
| | - Cong Shen
- Center for Reproduction and Genetics, Suzhou Municipal Hospital, the Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou
| | - Hong Li
- Center for Reproduction and Genetics, Suzhou Municipal Hospital, the Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou
| | - Xia Chen
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
- Reproductive Sciences Institute of Jiangsu University, Jiangsu University, Zhenjiang, China
| | - Jun Yu
- Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang
- Reproductive Sciences Institute of Jiangsu University, Jiangsu University, Zhenjiang, China
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13
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Funghi L, Torricelli M, Novembri R, Vannuccini S, Cevenini G, Di Tommaso M, Severi FM, Petraglia F. Placental and maternal serum activin A in spontaneous and induced labor in late-term pregnancy. J Endocrinol Invest 2018; 41:171-177. [PMID: 28612286 DOI: 10.1007/s40618-017-0640-z] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/19/2017] [Accepted: 02/14/2017] [Indexed: 11/29/2022]
Abstract
PURPOSE Feto-placental unit represents an important source of activin A, a member of transforming growth factors-β involved in the mechanisms of labor. No evidences are available on activin A in pregnancies beyond 41 weeks of gestation, where induction of labor is often required. The present study aimed to evaluate activin A maternal serum levels and placental mRNA expression in term and late-term pregnancy, with spontaneous or induced labor, and its possible role to predict the response to labor induction. METHODS Maternal serum samples and placental specimens were collected from women with singleton pregnancy admitted for either term spontaneous labor (n = 23) or induction of labor for late-term pregnancy (n = 41), to evaluate activin A serum levels and placental mRNA expression. Univariate and multivariate analyses on activin A serum levels, maternal clinical parameters, and cervical length were conducted in women undergoing induction of labor. RESULTS Maternal serum activin A levels and placental activin A mRNA expression in late-term pregnancies were significantly higher than at term. Late-term pregnancies who did not respond to induction of labor showed significantly lower levels of activin A compared to responders. The combination of serum activin A and cervical length achieved a sensitivity of 100% and a specificity of 93.55% for the prediction of successful induction. CONCLUSION Late-term pregnancy is characterized by hyperexpression of placental activin A and increased maternal activin A secretion. By combining maternal serum activin A levels with cervical length, a good predictive model for the response to induction of labor was elaborated.
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Affiliation(s)
- L Funghi
- Obstetrics and Gynecology, Department of Molecular and Developmental Medicine, University of Siena, Policlinico "Santa Maria alle Scotte" Viale Bracci, 53100, Siena, Italy
| | - M Torricelli
- Obstetrics and Gynecology, Department of Molecular and Developmental Medicine, University of Siena, Policlinico "Santa Maria alle Scotte" Viale Bracci, 53100, Siena, Italy
| | - R Novembri
- Obstetrics and Gynecology, Department of Molecular and Developmental Medicine, University of Siena, Policlinico "Santa Maria alle Scotte" Viale Bracci, 53100, Siena, Italy
| | - S Vannuccini
- Obstetrics and Gynecology, Department of Molecular and Developmental Medicine, University of Siena, Policlinico "Santa Maria alle Scotte" Viale Bracci, 53100, Siena, Italy
| | - G Cevenini
- Department of Medical Biotechnology, University of Siena, Siena, Italy
| | - M Di Tommaso
- Department of Health Sciences, University of Florence, Florence, Italy
| | - F M Severi
- Obstetrics and Gynecology, Department of Molecular and Developmental Medicine, University of Siena, Policlinico "Santa Maria alle Scotte" Viale Bracci, 53100, Siena, Italy
| | - F Petraglia
- Obstetrics and Gynecology, Department of Molecular and Developmental Medicine, University of Siena, Policlinico "Santa Maria alle Scotte" Viale Bracci, 53100, Siena, Italy.
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14
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Fan J, Yu S, Cui Y, Xu G, Wang L, Pan Y, He H. Bcl-2/Bax protein and mRNA expression in yak (Bos grunniens) placentomes. Theriogenology 2017; 104:23-29. [DOI: 10.1016/j.theriogenology.2017.07.045] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/30/2017] [Revised: 07/11/2017] [Accepted: 07/28/2017] [Indexed: 01/01/2023]
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15
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Xia S, Zhen Y, Ma H, Wang A. Abnormal expression of microRNA-575 leads to missed abortion through regulating apoptosis and angiogenesis. Exp Ther Med 2017; 14:3993-4000. [PMID: 29104621 PMCID: PMC5658693 DOI: 10.3892/etm.2017.5086] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/17/2016] [Accepted: 03/31/2017] [Indexed: 01/08/2023] Open
Abstract
Numerous microRNA (miR) are important for placental development and function. miR-575 has been demonstrated to be upregulated in maternal placenta in patients who have experienced a miscarriage. The present study aimed to explore the role of abnormal expression of miR-575 in missed abortion (MA) and to further analyze the potential molecular mechanisms. Embryo villus tissue samples were extracted from 10 childless women with MA and 10 fertile women without a history of MA. Additionally, human choriocarcinoma cells, JEG-3, were used in the present study, which were transfected with miR-575 mimic, inhibitor and scramble. The expression of miR-575 in embryo villus tissues and in JEG-3 cells was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Apoptosis in villus tissues of patients with MA and in JEG-3 cells of miR-575 mimic, inhibitor and scramble groups were detected by flow cytometry. Furthermore, the expression levels of apoptosis-related proteins, including B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) and phosphorylated (p)-p53, and angiogenesis-related proteins, including vascular endothelial growth factor (VEGF) and angiopoietin 2 (Ang-2), were measured by RT-qPCR and western blotting. Additionally, the target of miR-575 was predicted and clarified by luciferase reporter assay. miR-575 was significantly overexpressed in MA villus tissue compared with normal tissue (P<0.05). The percentage of apoptotic cells in MA embryo villus tissue was significantly higher than that in normal tissue (P<0.05). After JEG-3 cells were transfected with miR-575 inhibitor, the expression of miR-575 and the percentage of apoptotic cells decreased significantly compared with the control (P<0.05). MiR-575 suppression significantly increased the expression of Bcl-2 (P<0.05), and decreased the expressions of Bax (P<0.05) and p-p53 (P<0.01) compared with the control. Furthermore, miR-575 suppression significantly increased the expressions of angiogenesis-related proteins, Ang-2 and VEGF (P<0.01). Superoxide dismutase 2 was identified as the target of miR-575. Therefore, abnormal expression of miR-575 may lead to MA through regulating apoptosis and angiogenesis. Inhibition of miR-575 may inhibit apoptosis and promote angiogenesis in MA.
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Affiliation(s)
- Shuqin Xia
- Department of Gynecology and Obstetrics, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.,Department of Gynecology and Obstetrics, Inner Mongolia People's Hospital, Hohhot, Inner Mongolia 010017, P.R. China.,Department of Gynecology and Obstetrics, Navy General Hospital PLA China, Beijing 100048, P.R. China
| | - Yihui Zhen
- Department of Gynecology and Obstetrics, Navy General Hospital PLA China, Beijing 100048, P.R. China
| | - Hongsheng Ma
- Department of Gynecology and Obstetrics, Navy General Hospital PLA China, Beijing 100048, P.R. China
| | - Aiming Wang
- Department of Gynecology and Obstetrics, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.,Department of Gynecology and Obstetrics, Inner Mongolia People's Hospital, Hohhot, Inner Mongolia 010017, P.R. China
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16
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Chromium VI - Induced developmental toxicity of placenta is mediated through spatiotemporal dysregulation of cell survival and apoptotic proteins. Reprod Toxicol 2016; 68:171-190. [PMID: 27443218 DOI: 10.1016/j.reprotox.2016.07.006] [Citation(s) in RCA: 41] [Impact Index Per Article: 4.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2016] [Revised: 07/06/2016] [Accepted: 07/09/2016] [Indexed: 12/22/2022]
Abstract
Environmental contamination with hexavalent chromium (CrVI) is a growing problem both in the U.S and developing countries. CrVI is a heavy-metal endocrine disruptor; women working in Cr industries exhibit an increased incidence of premature abortion and infertility. The current study was designed to understand the mechanism of CrVI toxicity on placental cell survival/death pathways. Pregnant mothers were treated with or without CrVI (50ppmK2Cr2O7) through drinking water from gestational day (GD) 9.5-14.5, and placentas were analyzed on GD 18.5. Results indicated that CrVI increased apoptosis of trophoblasts, vascular endothelium of the metrial glands and yolk sac epithelium through caspase-3 and p53-dependent pathways. CrVI increased apoptosis in labyrinth and basal zones in a caspase-3-independent manner via AIF, and through an ATM-p53-NOXA-PUMA-p27 network. CrVI downregulated cell survival proteins Bcl-2, Bcl-XL and XIAP in the placenta. CrVI disrupts placental histoarchitecture and increases cell death by spatiotemporal modulation of apoptotic signaling.
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17
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Mastrolia SA, Weintraub AY, Sciaky-Tamir Y, Tirosh D, Loverro G, Hershkovitz R. Placental calcifications: a clue for the identification of high-risk fetuses in the low-risk pregnant population? J Matern Fetal Neonatal Med 2015; 29:921-7. [DOI: 10.3109/14767058.2015.1023709] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
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Aires MB, Santos ACVD. Effects of maternal diabetes on trophoblast cells. World J Diabetes 2015; 6:338-344. [PMID: 25789116 PMCID: PMC4360428 DOI: 10.4239/wjd.v6.i2.338] [Citation(s) in RCA: 30] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/14/2014] [Revised: 11/15/2014] [Accepted: 12/31/2014] [Indexed: 02/05/2023] Open
Abstract
Diabetes mellitus (DM) is a health condition characterized by hyperglycemia over a prolonged period. There are three main types of DM: DM type 1 (DM1), DM2 and gestational DM (GDM). Maternal diabetes, which includes the occurrence of DM1 and DM2 during pregnancy or GDM, increases the occurrence of gesttional complications and adverse fetal outcomes. The hyperglycemic intrauterine environment affects not only the fetus but also the placental development and function in humans and experimental rodents. The underlying mechanisms are still unclear, but some evidence indicates alterations in trophoblast proliferation, apoptosis and cell cycle control in diabetes. A proper coordination of trophoblast proliferation, differentiation and invasion is required for placental development. Initially, increased expression of proliferative markers in junctional and labyrinth zones of rat placentas and villous cytotrophoblast, syncytiotrophoblast, stromal cells and fetal endothelial cells in human placentas is reported among diabetics. Moreover, reduced apoptotic index and expression of some apoptotic genes are described in placentas of GDM women. In addition, cell cycle regulators including cyclins and cyclin-dependent kinase inhibitors seem to be affected by the hyperglycemic environment. More studies are necessary to check the balance between proliferation, apoptosis and differentiation in trophoblast cells during maternal diabetes.
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Zhang Z, Yang X, Zhang L, Duan Z, Jia L, Wang P, Shi Y, Li Y, Gao J. Decreased expression and activation of Stat3 in severe preeclampsia. J Mol Histol 2015; 46:205-19. [DOI: 10.1007/s10735-015-9613-8] [Citation(s) in RCA: 28] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2014] [Accepted: 03/03/2015] [Indexed: 01/09/2023]
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20
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Chen H, Deng X, Yang Y, Shen Y, Chao L, Wen Y, Sun Y. Expression of GRIM-19 in missed abortion and possible pathogenesis. Fertil Steril 2015; 103:138-46.e3. [DOI: 10.1016/j.fertnstert.2014.10.012] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/05/2014] [Revised: 09/21/2014] [Accepted: 10/09/2014] [Indexed: 01/20/2023]
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21
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Modulation of wolframin expression in human placenta during pregnancy: comparison among physiological and pathological states. BIOMED RESEARCH INTERNATIONAL 2014; 2014:985478. [PMID: 24588001 PMCID: PMC3920918 DOI: 10.1155/2014/985478] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 04/29/2013] [Revised: 10/24/2013] [Accepted: 11/06/2013] [Indexed: 01/10/2023]
Abstract
The WFS1 gene, encoding a transmembrane glycoprotein of the endoplasmic reticulum called wolframin, is mutated in Wolfram syndrome, an autosomal recessive disorder defined by the association of diabetes mellitus, optic atrophy, and further organ abnormalities. Disruption of the WFS1 gene in mice causes progressive β-cell loss in the pancreas and impaired stimulus-secretion coupling in insulin secretion. However, little is known about the physiological functions of this protein. We investigated the immunohistochemical expression of wolframin in human placenta throughout pregnancy in normal women and diabetic pregnant women. In normal placenta, there was a modulation of wolframin throughout pregnancy with a strong level of expression during the first trimester and a moderate level in the third trimester of gestation. In diabetic women, wolframin expression was strongly reduced in the third trimester of gestation. The pattern of expression of wolframin in normal placenta suggests that this protein may be required to sustain normal rates of cytotrophoblast cell proliferation during the first trimester of gestation. The decrease in wolframin expression in diabetic placenta suggests that this protein may participate in maintaining the physiologic glucose homeostasis in this organ.
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22
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Fraga LR, Dutra CG, Boquett JA, Vianna FSL, Gonçalves RO, Paskulin DD, Costa OL, Ashton-Prolla P, Sanseverino MTV, Schuler-Faccini L. p53 signaling pathway polymorphisms associated to recurrent pregnancy loss. Mol Biol Rep 2014; 41:1871-7. [PMID: 24435975 DOI: 10.1007/s11033-014-3036-6] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/12/2013] [Accepted: 01/04/2014] [Indexed: 01/24/2023]
Abstract
The p53 protein is known for performing essential functions in the maintenance of genomic stability in somatic cells and prevention of tumor formation. Studies of the p53 signaling pathway have suggested associations between some polymorphisms and infertility, post-in vitro fertilization implantation failure and recurrent abortions. The TP53 Pro72Arg polymorphism has been implicated as a risk factor for recurrent pregnancy loss (RPL); however, the association is controversial. In this study, our objective was to evaluate selected polymorphisms in genes of the p53 signalling pathway [TP53 c.215G>C (Pro72Arg), MDM2 c.14+309T>G (SNP309) and LIF c.1414T>G in the region 3' UTR] and determine their effect as risk factors for RPL. In a case-control study, we investigated 120 women with two or more pregnancy losses and 143 fertile control women reporting at least two live births and no history of pregnancy loss. When analyzed separately, the allele and genotype distributions of the polymorphisms in the two groups were not different. However, in a multivariate analysis adjusted for alcohol consumption, smoking, ethnicity, and number of pregnancies, the interaction between the genotypes TP53 Arg/Arg (rs1042522) and MDM2 TT (rs2279744) showed to be associated to RPL, increasing the risk for this condition (OR = 2.58, 95% CI: 1.31-5.07, p = 0.006). In conclusion, our study indicates that the combination of TP53 Arg/Arg (rs1042522) and MDM2 TT (rs2279744) genotypes may be a risk factor for RPL.
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Affiliation(s)
- L R Fraga
- Post-Graduation Program in Genetics and Molecular Biology, Departament of Genetics, Biosciences Institute, Universidade Federal do Rio Grande do Sul (UFRGS), Caixa Postal 15031 - Agencia Campus UFRGS, Porto Alegre, RS, 91501-970, Brazil,
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23
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Wei D, Wu Q, Shi H. Apoptosis and p53 expression in the placental villi of females with unexplained recurrent spontaneous abortion. Exp Ther Med 2013; 7:191-194. [PMID: 24348788 PMCID: PMC3861175 DOI: 10.3892/etm.2013.1399] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/04/2013] [Accepted: 10/28/2013] [Indexed: 01/17/2023] Open
Abstract
The aim of this study was to explore the level of apoptosis and p53 expression in the placental villi of patients with unexplained recurrent spontaneous abortion (URSA). Fifty-three pregnant females with URSA and 32 pregnant females who required an induced abortion were selected as the subjects of this study. Placental villus tissues were collected from June 2010 to June 2012 and quantitative polymerase chain reaction (qPCR) and immunohistochemical analysis were performed to determine the mRNA and protein levels of p53 in the placental villus tissues. The level of apoptosis in the tissues was studied using terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) assay. The mRNA and protein expression levels of p53 in the URSA group were significantly higher than those in the control group (P<0.05). Furthermore, the levels of apoptosis were increased markedly in the URSA group compared with the control group (P<0.05). In conclusion, the placental villi of patients with URSA express a high level of p53, which may result in cell apoptosis and lead to recurrent spontaneous abortion.
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Affiliation(s)
- Dehua Wei
- Department of Gynecology and Obstetrics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450000, P.R. China ; Department of Gynecology and Obstetrics, Puyang People's Hospital, Puyang, Henan 457000, P.R. China
| | - Qinghua Wu
- Department of Gynecology and Obstetrics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450000, P.R. China
| | - Huirong Shi
- Department of Gynecology and Obstetrics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450000, P.R. China
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24
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Sharma M, Kumar R, Bhatla N, Dhingra R. A comparative study of apoptosis in placentas of normal and preeclamptic Indian pregnant women by TUNEL assay and M30 immunostaining. J Clin Lab Anal 2013; 26:459-66. [PMID: 23143629 DOI: 10.1002/jcla.21547] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/02/2023] Open
Abstract
BACKGROUND Preeclampsia is a unique life-threatening disorder of human pregnancy associated with the abnormal placentation caused by the inadequate trophoblastic invasion due to altered apoptosis of these cells. The aim of the present study was to assess and compare the apoptosis in trophoblastic cells in various zones (villous and extravillous) of placentas of preeclamptic and normotensive nonproteinuric pregnant women. METHODS Hematoxylin eosin staining, TUNEL assay and M30 immunostaining techniques were used for studying apoptosis in trophoblastic cells of placentas of two groups. The results of apoptotic indices by these techniques were compared in preeclamptic group. RESULTS The TUNEL apoptotic indices were higher in all the zones of placentas of preeclamptic group as compared to control group though the results were not statistically significant. M30 immunostaining also gave higher apoptotic indices in all the zones of preeclamptic placentas as compared to the normal group but the result of apoptotic index of basal plate was not statistically significant. M30 immunostaining was absent in stromal cells. CONCLUSION In our study, the trophoblastic apoptotic rates were more in villous as well as extravillous zones of preeclamptic placentas as compared to normal placentas and M30 immunostaining was found to be more sensitive and specific for the apoptotic trophoblastic cells as compared to TUNEL assay.
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Affiliation(s)
- Mona Sharma
- Department of Anatomy, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.
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25
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Börzsönyi B, Demendi C, Rigó J, Szentpéteri I, Rab A, Joó JG. The regulation of apoptosis in intrauterine growth restriction: a study ofBcl-2andBaxgene expression in human placenta. J Matern Fetal Neonatal Med 2012; 26:347-50. [DOI: 10.3109/14767058.2012.733770] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
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Demendi C, Börzsönyi B, Végh V, Nagy ZB, Rigó J, Pajor A, Joó JG. Gene expression patterns of the Bcl-2 and Bax genes in preterm birth. Acta Obstet Gynecol Scand 2012; 91:1212-7. [PMID: 22524261 DOI: 10.1111/j.1600-0412.2012.01428.x] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/15/2023]
Abstract
OBJECTIVE The apoptotic genes Bax and Bcl-2 are both involved in the pathogenesis of preterm delivery in conjunction with additional factors. We characterized gene expression patterns of these apoptotic regulatory genes as well as relevant environmental factors. DESIGN A gene expression study with evaluation of clinical data. SETTING Semmelweis University, Budapest, Hungary. SAMPLE Human placental samples from 104 preterm and 140 full-term pregnancies. METHODS Gene tests were performed using real-time PCR to assess gene expression patterns of Bax and Bcl-2 in human placental samples. Clinical data were collected from our computerized database. MAIN OUTCOME MEASURES Apoptotic gene expression pattern and clinical information against the background of preterm delivery. RESULTS In placental samples from preterm delivery pregnancies, expression of the Bcl-2 gene was unchanged, whereas the Bax gene was overexpressed. Placental gene expression of Bax in preterm delivery was dependent on gestational age with gestational weeks 28-32 and 32-36 associated with overexpression, and no overexpression in gestational weeks 24-28. Preterm delivery began with premature rupture of membranes in 70.2% and spontaneous uterine activity in 29.8%. CONCLUSIONS The Bax gene was overexpressed in preterm delivery, whereas expression of the Bcl-2 gene remained unchanged. After the 28(th) gestational week, apoptosis appears to be a key factor in the pathogenesis of preterm delivery.
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Affiliation(s)
- Csaba Demendi
- Second Department of Gynecology and Obstetrics, Semmelweis University, Budapest, Hungary
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Soni S, Rath G, Prasad CP, Salhan S, Saxena S, Jain AK. Apoptosis and Bcl-2 protein expression in human placenta over the course of normal pregnancy. Anat Histol Embryol 2012; 39:426-31. [PMID: 20608924 DOI: 10.1111/j.1439-0264.2010.01012.x] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
Apoptosis plays a central role in organ development, homeostasis and immune defence in multicellular organisms and is strictly controlled in part by members of Bcl-2 family. The Bcl-2 is a pro-survival molecule identified through its involvement in B-cell lymphomas. The aim of the study was to evaluate the incidence of apoptosis in the human placenta at different stages of pregnancy and to correlate it further with Bcl-2 expression. A total of 96 placental samples from first trimester, mid-trimester and uncomplicated term pregnancies were collected (n = 32 + 32 + 32). M30 cyto death monoclonal antibody was used to identify apoptotic cells. The apoptosis index of first trimester placentae was 2.33 ± 1.70, mid- trimester was 1.77 ± 1.36 and term placenta was 1.15 ± 0.21. Bcl-2 protein was found immunolocalized in the cytoplasm of syncytiotrophoblast. Apoptosis index was significantly reduced in term cases as compared with first trimester (P < 0.002) and mid-trimester placentae (P = 0.01). On the contrary, Bcl-2 expression was significantly higher at term cases than in first trimester (P < 0.0001) and mid-trimester cases (P < 0.001). The present study divulges the importance of apoptosis in permitting normal physiological turnover of villous trophoblast and also exhibits the contribution of bcl-2 in maintaining syncytial integrity throughout normal pregnancy.
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Affiliation(s)
- S Soni
- Department of Anatomy, Vardhman Mahavir Medical College & Safdarjung Hospital, New Delhi, India
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28
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Hills FA, Mehmet H, Sullivan MH. Insulin-like growth factor-II and heparin are anti-apoptotic survival factors in human villous cytotrophoblast. Eur J Obstet Gynecol Reprod Biol 2012; 163:11-6. [DOI: 10.1016/j.ejogrb.2012.03.013] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/08/2011] [Revised: 11/22/2011] [Accepted: 03/08/2012] [Indexed: 10/28/2022]
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Torricelli M, Novembri R, Conti N, De Falco G, De Bonis M, Petraglia F. Correlation With Placental Kisspeptin in Postterm Pregnancy and Apoptosis. Reprod Sci 2012; 19:1133-7. [DOI: 10.1177/1933719112443878] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Affiliation(s)
- Michela Torricelli
- Department of Pediatrics, Obstetrics and Reproductive Medicine, Division of Obstetrics and Gynecology, University of Siena, Siena, Italy
| | - Romina Novembri
- Department of Pediatrics, Obstetrics and Reproductive Medicine, Division of Obstetrics and Gynecology, University of Siena, Siena, Italy
| | - Nathalie Conti
- Department of Pediatrics, Obstetrics and Reproductive Medicine, Division of Obstetrics and Gynecology, University of Siena, Siena, Italy
| | - Giulia De Falco
- Department of Human Pathology and Oncology, University of Siena, Siena, Italy
| | - Maria De Bonis
- Department of Pediatrics, Obstetrics and Reproductive Medicine, Division of Obstetrics and Gynecology, University of Siena, Siena, Italy
| | - Felice Petraglia
- Department of Pediatrics, Obstetrics and Reproductive Medicine, Division of Obstetrics and Gynecology, University of Siena, Siena, Italy
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Fang Y, Kong B, Yang Q, Ma D, Qu X. The p53-HDM2 gene-gene polymorphism interaction is associated with the development of missed abortion. Hum Reprod 2011; 26:1252-8. [DOI: 10.1093/humrep/der017] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/25/2022] Open
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Fang Y, Kong B, Yang Q, Ma D, Qu X. MDM2 309 polymorphism is associated with missed abortion. Hum Reprod 2009; 24:1346-9. [PMID: 19246469 DOI: 10.1093/humrep/dep044] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
BACKGROUND In this study, we assessed whether the single nucleotide polymorphism in the murine double minute 2 (MDM2) promoter (SNP309) was associated with the occurrence of missed abortion. METHODS Genotyping of MDM2 SNP309 polymorphism was conducted by polymerase chain reaction-restriction fragment length polymorphism with blood and villous samples from 95 women diagnosed as having 1st trimester missed abortion. RESULTS The MDM2 SNP309 G/G genotype was associated with a higher risk of missed abortion compared with the T/T+ T/G genotype in blood (P = 0.010; odds ratio (OR): 2.164; 95% confidence interval (CI): 1.207-3.878) and villous samples (P = 0.043; OR: 2.767; 95% CI: 1.092-7.011). CONCLUSIONS The MDM2 SNP309 G/G genotype may be a genetic risk factor for missed abortion.
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Affiliation(s)
- Yan Fang
- Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, 107 Wenhuaxi Road, Ji'nan 250012, Shandong, People's Republic of China
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Yamauchi H, Katayama KI, Ueno M, Kanemitsu H, Nam C, Mikami T, Saito A, Ishida Y, Uetsuka K, Doi K, Ohmach Y, Nakayama H. Etoposide induces TRP53-dependent apoptosis and TRP53-independent cell cycle arrest in trophoblasts of the developing mouse placenta. Biol Reprod 2008; 80:813-22. [PMID: 19109225 DOI: 10.1095/biolreprod.108.069419] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023] Open
Abstract
Abnormal regulation of placental apoptosis and proliferation has been implicated in placental disorders. Recently, several DNA-damaging agents were reported to induce excessive apoptosis and reduce cell proliferation in the placenta; however, the molecular pathways of these toxic effects on the placenta are unclear. The aim of the present study was to determine the involvement of TRP53, a tumor suppressor that mediates cellular responses to DNA damage, in the induction of apoptosis and cell cycle arrest in the developing placenta. For this purpose, we treated pregnant mice on Day 12 of gestation with 10 mg/kg of etoposide and 5-Gy gamma irradiation, potent inducers of DNA damage. We found an increase in the number of trophoblastic apoptoses 8 and 24 h after etoposide injection and 6 and 24 h after irradiation in the placental labyrinth zone. The number of mitoses and DNA syntheses in trophoblasts decreased after treatment. The accumulation and phosphorylation of TRP53 protein were detected 8 and 6 h after etoposide injection and irradiation, respectively. In Trp53-deficient placentas, the induction of etoposide-induced trophoblastic apoptosis is abrogated, while the reduction of proliferation occurred similarly as in wild-type placentas. CDC2A, a regulator of G2/M progression, was inactivated by phosphorylation after etoposide injection and irradiation, suggesting that the cell cycle was arrested at the G2/M border by treatment. Our study demonstrated that etoposide injection induced TRP53-dependent apoptosis and TRP53-independent cell cycle arrest in labyrinthine trophoblasts, providing insights into the molecular pathway of placental disorders.
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Affiliation(s)
- Hirofumi Yamauchi
- Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
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Daher S, Guimarães AJ, Mattar R, Ishigai MMS, Barreiro EG, Bevilacqua E. Bcl-2 and Bax expressions in pre-term, term and post-term placentas. Am J Reprod Immunol 2008; 60:172-8. [PMID: 18705844 DOI: 10.1111/j.1600-0897.2008.00609.x] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022] Open
Abstract
PROBLEM Placental apoptosis-associated protein imbalance might contribute to the pathogenesis of pre- and post-term birth. Therefore, we evaluated the expression and distribution of pro-apoptotic (Bax) and anti-apoptotic (Bcl-2) molecules in term, pre-term and post-term placentas. METHOD OF STUDY Placental samples were collected from women with term (n = 25), pre-term (n = 7) and post-term (n = 10) deliveries. The expression of Bcl-2 and Bax was assessed by immunochemistry on paraffin-embedded placental specimens. RESULTS The pattern of immunostaining for Bcl-2 and Bax was the same in all samples, but not the intensity. The Bax/Bcl-2 ratio was higher in both pre-term and post-term placental samples compared with term placentas as a result of intense reactivity for the pro-apoptotic factor, Bax in pre-term and post-term placentas and, for Bcl-2 decrease in pre-term placentas. CONCLUSION These findings suggest that different unbalance mechanisms in placental apoptotic-associated protein expressions may be involved in the physiopathology of pre-term and post-term births.
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Affiliation(s)
- Silvia Daher
- Department of Obstetrics, Paulista Medical School, São Paulo Federal University, São Paulo, Brazil.
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Nakashima A, Shiozaki A, Myojo S, Ito M, Tatematsu M, Sakai M, Takamori Y, Ogawa K, Nagata K, Saito S. Granulysin produced by uterine natural killer cells induces apoptosis of extravillous trophoblasts in spontaneous abortion. THE AMERICAN JOURNAL OF PATHOLOGY 2008; 173:653-64. [PMID: 18688023 DOI: 10.2353/ajpath.2008.071169] [Citation(s) in RCA: 75] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
Immune changes are known to occur in recurrent spontaneous abortion, but it is unclear whether either maternal natural killer (NK) cells or T cells attack fetus-derived trophoblasts. To clarify the immunological causes of spontaneous abortion, we examined the relationship between cytotoxic granule proteins in decidual lymphocytes, such as granulysin, granzyme B, and perforin, and the induction of apoptosis in extravillous trophoblasts (EVTs). The number of granulysin-positive CD56(bright) NK cells increased significantly in the decidua basalis during spontaneous abortion compared with normal pregnancy; however, granzyme B- and perforin-positive cells did not change. Interestingly, the expression of granulysin was also detected in the nuclei of EVTs in spontaneous abortion samples. When IL-2-stimulated CD56(bright) NK cells were cocultured with EVT cells (HTR-8/SV40neo), granulysin was found initially in the cytoplasm and then accumulated in the nuclei of the HTR-8/SV40neo cells. Furthermore, transfected cells expressing a GFP-granulysin fusion protein induced apoptosis in HTR-8/SV40neo cells independently of caspases. Our results suggest that granulysin-positive uterine NK cells attack EVTs; subsequently, the uNK-derived granulysin actively accumulates in the nuclei of EVTs, causing the death of EVTs due to apoptosis. These data support a new apoptosis pathway for trophoblasts via uNK-derived granulysin, suggesting that granulysin is involved in spontaneous abortion.
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Affiliation(s)
- Akitoshi Nakashima
- Department of Obstetrics and Gynecology, Faculty of Medicine, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan
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Guvendag Guven ES, Okur H, Beksac MS. ORIGINAL ARTICLE: Placental Fas/Fas Ligand Expression in Early Pregnancy Losses. Am J Reprod Immunol 2008; 60:1-7. [DOI: 10.1111/j.1600-0897.2008.00583.x] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022] Open
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Yamauchi H, Katayama KI, Ueno M, He XJ, Mikami T, Uetsuka K, Doi K, Nakayama H. Essential role of p53 in trophoblastic apoptosis induced in the developing rodent placenta by treatment with a DNA-damaging agent. Apoptosis 2008; 12:1743-54. [PMID: 17594519 DOI: 10.1007/s10495-007-0099-z] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/23/2022]
Abstract
Placental apoptosis plays important roles in both normal morphogenesis and pathogenesis. We previously reported that administration of cytosine arabinoside (Ara-C), a DNA-damaging agent, to pregnant rats induced apoptosis of trophoblasts in the placental labyrinth zone. Our aim here was to clarify the molecular pathway of DNA damage induced-trophoblastic apoptosis. We found the accumulation and phosphorylation of p53 protein, a tumor suppressor that mediates apoptosis under various cellular stresses, in Ara-C-treated rat placentas. Expression of the mRNAs of downstream targets of p53 was upregulated, suggesting that p53 exerts its function as a transcription factor. We also observed release of mitochondrial cytochrome c and activation of caspase-9, hallmarks of the intrinsic apoptotic pathway. Phosphorylation of Chk1 and H2A.X, target substrates of DNA damage transducers, was detected immediately after Ara-C treatment, suggesting activation of DNA damage cascades to phosphorylate p53. Ara-C-induced trophoblastic apoptosis was almost completely abrogated in placentas of Trp53 (coding p53)-deficient mice, whereas the levels of physiological apoptosis in trophoblasts were similar among wild-type and Trp53-deficient mice. These results indicate that p53 is essential for DNA damage-induced trophoblastic apoptosis and suggest that the mechanisms that regulate the damage-induced apoptosis differ from those that regulate physiological apoptosis.
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Affiliation(s)
- Hirofumi Yamauchi
- Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
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Abstract
Infantile haemangioma is the most common tumour of infancy, yet the origin of these lesions remains controversial and the predictable life cycle is poorly understood. Much new information on infantile haemangiomas has emerged over the past decade, but experts continue to debate fundamental features, including cell of origin, nonrandom distribution, and mechanisms regulating the sometimes explosive growth and slow involution. The development of useful laboratory models has been difficult, in turn restricting the development of treatment options available to the clinician. Despite this, new research and creative thinking has spawned several hypotheses on the origin of these tumours and their interesting clinical behaviour, including suggestions of an intrinsic defect in local endothelial cells, a contribution of circulating endothelial progenitors or haemangioblasts, embolisation of shed placental cells and developmental field defects. While no single hypothesis seems to describe all features of infantile haemangioma, continued research seeks to integrate these ideas, create a better understanding of these important tumours and bring new treatments to the clinic.
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Tjoa ML, Cindrova-Davies T, Spasic-Boskovic O, Bianchi DW, Burton GJ. Trophoblastic oxidative stress and the release of cell-free feto-placental DNA. THE AMERICAN JOURNAL OF PATHOLOGY 2006; 169:400-4. [PMID: 16877342 PMCID: PMC1698796 DOI: 10.2353/ajpath.2006.060161] [Citation(s) in RCA: 161] [Impact Index Per Article: 8.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
Considerable quantities of cell-free fetal DNA circulate in the maternal blood during human pregnancy, but the origin of the DNA remains uncertain. Circumstantial evidence suggests the placenta is the principal source, so we tested the hypothesis that release occurs from the syncytiotrophoblast after the induction of apoptotic changes. Villous explants from normal placentas delivered by elective caesarean section were cultured under normoxic conditions (10% oxygen) for up to 20 hours or exposed to hypoxia (0.5% oxygen) for 1 hour followed by reoxygenation. The concentration of beta-globin cell-free DNA in the supernatant, measured using real-time polymerase chain reaction methodology, was significantly increased at 20 hours after hypoxia-reoxygenation. Release was associated with increased apoptosis, confirmed by increased activation of caspase-3 on Western blotting, and immunolocalized to the syncytiotrophoblast; necrosis was also evidenced by release of lactate dehydrogenase. Both release of cell-free DNA and apoptosis could be significantly reduced by the addition of antioxidant vitamins C and E to the culture medium. This study provides the first evidence of a mechanistic and quantitative link between placental apoptosis/necrosis and release of cell-free DNA, hence confirming that maternal serum/plasma concen-trations of cell-free DNA may act as a biomarker of trophoblast well-being during pregnancy.
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Affiliation(s)
- May Lee Tjoa
- Department of Pediatrics, Division of Genetics, Tufts-New England Medical Center, Boston, Massachusetts, USA
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Sgarbosa F, Barbisan LF, Brasil MAM, Costa E, Calderon IMP, Gonçalves CR, Bevilacqua E, Rudge MVC. Changes in apoptosis and Bcl-2 expression in human hyperglycemic, term placental trophoblast. Diabetes Res Clin Pract 2006; 73:143-9. [PMID: 16563550 DOI: 10.1016/j.diabres.2005.12.014] [Citation(s) in RCA: 51] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/30/2005] [Revised: 12/05/2005] [Accepted: 12/22/2005] [Indexed: 12/25/2022]
Abstract
Apoptosis and its associated regulatory mechanisms are physiological events crucial to the maintenance of placental homeostasis; imbalance of these processes, however, such as occurs under various pathological conditions, may compromise placenta function and, consequently, pregnancy success. Increased apoptosis occurs in the placentas of pregnant women with several developmental disabilities, while increased Bcl-2 expression is generally associated with pregnancy-associated tumors. Herein, we tested the hypothesis that apoptosis-associated disturbs might be involved in the placental physiopathology subjected to different maternal hyperglycemic conditions. Thus, in the present study we investigated and compared the incidence of apoptosis using TUNEL reaction and Bcl-2 expression, in term-placentas of normoglycemic, diabetic and daily hyperglycemic patients. Tissue samples were collected from 37 placentas, being 15 from healthy mothers with normally delivered healthy babies, and 22 from mothers with glucose disturbances. From these latter 22 patients, 10 showed maternal daily hyperglycemia and 12 were clinically diabetics. Both Bcl-2 expression and apoptotic DNA fragmentation were established and quantified in the trophoblasts of healthy mothers. Compared to these reference values, a higher apoptosis index and lower Bcl-2 expression were disclosed in the placentas of the diabetic women, while in the daily hyperglycemic group, values were intermediate between the diabetic and normoglycemic patients. The TUNEL/Bcl-2 index ratio in the placentas varied from 0.02 to 0.09 for pregnant normoglycemic and diabetic women, respectively, revealing a predominance of apoptosis in the diabetic group. Our findings suggest that hyperglycemia may be a key factor evoking apoptosis in the placental trophoblast, and therefore, is relevant to diabetic placenta function.
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Affiliation(s)
- Fabio Sgarbosa
- Department of Gynecology and Obstetrics, School of Medicine of Botucatu, UNESP - São Paulo State University, 18618-000 Botucatu, SP, Brazil
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Toledo MT, Ventrucci G, Marcondes MCCG. Cancer during pregnancy alters the activity of rat placenta and enhances the expression of cleaved PARP, cytochrome-c and caspase 3. BMC Cancer 2006; 6:168. [PMID: 16800886 PMCID: PMC1534057 DOI: 10.1186/1471-2407-6-168] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2005] [Accepted: 06/26/2006] [Indexed: 02/08/2023] Open
Abstract
Background The presence of cancer makes it difficult to predict the progress of pregnancy and can be deleterious to the maternal-foetal relationship. Apoptosis may affect a range of placental functions and result in the retardation of foetal growth. In this work, we investigated the placental alterations produced by tumour growth and the effects on the expression of apoptotic factors in placental tissue. Methods Adult female Wistar rats (90 days old, n = 54) were allocated to control (C), tumour-bearing (W), or ascitic fluid-injected (A) groups and were killed on the 16th, 19th or 21st day of pregnancy. Placental tissues were analysed using biochemical and histochemical assays. Results The placental protein content and glutathione-S-transferase activity were decreased in groups W and A. Histochemical analysis showed an increase in the number of cells with cleaved PARP, caspase 3 and cytochrome-c in groups W and A, indicating that the tumour growth clearly damaged placental tissue and affected the levels of apoptotic factors. These results were confirmed by western blotting. Conclusion Since trophoblastic cells are responsible for maintaining a normal placental function, the uncontrolled death of these cells in response to tumour cell growth or substances derived from ascitic fluid could have a negative impact on foetal development. Further knowledge of these events may help to preserve the foetus and placenta during development.
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Affiliation(s)
- Mércia Tancredo Toledo
- Departamento de Fisiologia e Biofísica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), Campinas, CP 6109, 13083-970, SP, Brazil
| | - Gislaine Ventrucci
- Departamento de Fisiologia e Biofísica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), Campinas, CP 6109, 13083-970, SP, Brazil
| | - Maria Cristina Cintra Gomes Marcondes
- Departamento de Fisiologia e Biofísica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), Campinas, CP 6109, 13083-970, SP, Brazil
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Ganapathy R, Ayling LJ, Whitley GSJ, Cartwright JE, Thilaganathan B. Effect of first-trimester serum from pregnant women with high-resistance uterine artery Doppler resistance on extravillous trophoblast invasion. Hum Reprod 2006; 21:1295-8. [PMID: 16431899 DOI: 10.1093/humrep/dei482] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/14/2022] Open
Abstract
BACKGROUND Abnormal uterine artery Doppler indices are associated with pregnancy complications such as pre-eclampsia and intrauterine growth restriction. Poor trophoblast invasion may be a consequence of, or be associated with, abnormal Doppler indices. OBJECTIVE To evaluate in vitro trophoblast function following exposure to first-trimester serum from pregnancies with high uterine artery Doppler resistance indices. METHODS Doppler ultrasound examination of the maternal uterine arteries was performed on women at 10-14 weeks' gestation. Serum was collected from women with bilateral uterine artery notches with resistance indices above the 95th centile and from patients with normal uterine artery indices. The effect of serum on trophoblast invasion was determined using an established in vitro model from the extravillous trophoblast-derived cell line SGHPL-4. RESULTS Trophoblastic invasion was significantly reduced when treated with serum from women with high-resistance compared with normal-resistance uterine artery Doppler indices (P < 0.05). CONCLUSION Maternal serum in the first trimester of pregnancy from patients with high-resistance uterine artery Doppler indices appears to inhibit trophoblast invasion. This experimental model allows further investigation of factors responsible and the evaluation of therapeutic strategies.
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Affiliation(s)
- R Ganapathy
- Biochemistry and Immunology, Division of Basic Medical Sciences, Division of Clinical Developmental Sciences, St George's University of London, London, UK
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Dash PR, Whitley GSJ, Ayling LJ, Johnstone AP, Cartwright JE. Trophoblast apoptosis is inhibited by hepatocyte growth factor through the Akt and beta-catenin mediated up-regulation of inducible nitric oxide synthase. Cell Signal 2005; 17:571-80. [PMID: 15683732 DOI: 10.1016/j.cellsig.2004.09.015] [Citation(s) in RCA: 38] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/26/2004] [Revised: 09/13/2004] [Accepted: 09/20/2004] [Indexed: 12/18/2022]
Abstract
Excessive apoptosis of trophoblast cells is thought to be a contributing factor in complications of pregnancy such as pre-eclampsia. Hepatocyte growth factor (HGF) inhibits apoptosis in trophoblasts and we have investigated the signalling pathways through which this anti-apoptotic effect is mediated. Treatment of cells with HGF led to rapid phosphorylation of Akt while an Akt inhibitor blocked the protective effect of HGF. Glycogen synthase kinase-3beta (GSK-3beta) was found to be one of the downstream targets of Akt. HGF treatment inactivated GSK-3beta which in turn led to the activation of the transcription factor beta-catenin. Pharmacological inhibition of GSK-3beta, independently of HGF treatment, strongly increased both beta-catenin activity and cell survival, suggesting that beta-catenin alone has a pronounced anti-apoptotic effect. We also found that both HGF treatment and pharmacological activation of beta-catenin leads to increased expression of inducible nitric oxide synthase (iNOS). We suggest that the Akt mediated activation of beta-catenin leads to inhibition of trophoblast apoptosis following increased expression of iNOS.
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Affiliation(s)
- Philip R Dash
- Department of Basic Medical Sciences, Division of Biochemistry and Immunology, St. George's Hospital Medical School, University of London, Cranmer Terrace, London SW17 ORE, UK.
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Akirav C, Lu Y, Mu J, Qu DW, Zhou YQ, Slevin J, Holmyard D, Foster FS, Adamson SL. Ultrasonic detection and developmental changes in calcification of the placenta during normal pregnancy in mice. Placenta 2005; 26:129-37. [PMID: 15708114 DOI: 10.1016/j.placenta.2004.05.010] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Accepted: 05/19/2004] [Indexed: 11/29/2022]
Abstract
High resolution ultrasound imaging of the mouse placenta during development revealed highly echogenic foci localized near the materno-placental interface in early gestation and, near term, in the placental labyrinth (the exchange region of the placenta). Echogenic foci and calcium deposits identified in histological sections using Alizarin red staining showed similar localization and changes with gestation. Calcium deposits caused the echogenic foci because incubating uteri in a decalcifying solution eliminated both the deposits and echogenic foci. Transmission electron microscopy, X-ray microanalysis, and electron diffraction were used to show that deposits were calcium hydroxyapatite crystals. Calcium deposits were extensive and densely packed at days 7.5-9.5 of gestation at the border between the maternal decidua and the fetal trophoblast giant cells of ectoplacental cone. After the formation of the chorio-allantoic placenta (approximately day 10.5), calcification deposits appeared larger and more rarefied but were still localized at the border between the maternal decidua and the fetal trophoblast giant cells of the placenta. Calcification deposits were not observed in the labyrinthine region of the mouse placenta until > or = day 15.5 (day 18.5 is full term). We conclude that deposits of calcium hydroxyapatite crystals in the mouse placenta are detectable by high resolution ultrasound imaging. These deposits provide an ultrasound detectable marker of the maternal-placental interface that is particularly prominent during the establishment of the chorio-allantoic placenta between days 7.5 and 9.5 of gestation.
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Affiliation(s)
- C Akirav
- Samuel Lunenfeld Research Institute of Mount Sinai Hospital, Toronto, Ontario, Canada
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Austgulen R, Isaksen CV, Chedwick L, Romundstad P, Vatten L, Craven C. Pre-eclampsia: associated with increased syncytial apoptosis when the infant is small-for-gestational-age. J Reprod Immunol 2004; 61:39-50. [PMID: 14967220 DOI: 10.1016/j.jri.2003.10.001] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2003] [Revised: 10/20/2003] [Accepted: 10/20/2003] [Indexed: 10/26/2022]
Abstract
The present investigation was undertaken to study the association between placental apoptosis and pre-eclampsia, discriminating between pre-eclamptic pregnancies with appropriate-, and small-for-gestational-age (SGA), infants. Twenty pregnancies with pre-eclampsia and SGA (birth weight at or below -2 standard deviations) infants were selected in a retrospective study. Subsequently, corresponding numbers of gestational age-matched pre-eclampsia cases with appropriate-gestational-age (AGA) (birth weight at or above the 50% centile) infants and AGA controls without pre-eclampsia were selected. Formalin-fixed placental tissue was obtained from all groups. Apoptosis was assessed by a monoclonal antibody (M30), detecting a neoepitope of cytokeratin that is generated early in the apoptotic cascade. M30-positive cells were counted in villous and decidual/ basal plate tissue fields, and results were given as numbers of M30-positive cells per field. The study was performed blinded. Increased apoptosis was found in the syncytiotrophoblast layer in pre-eclampsia with SGA infants (0.14 apototic incidents per field of villous tissue, with 0.04-0.23 as the corresponding 25-75% inter quartile range (IQR) (P=0.05)). Syncytial apoptosis in the syncytial layer in the pre-eclampsia group with AGA infants was lower (0.09, IQR 0.03-0.15) and corresponded to the level detected among controls (0.06, IQR 0.03-0.17). Apoptosis in other placental cellular compartments did not differ between groups. The increased syncytial apoptosis found in placentas from pregnancies with SGA infants may either be due to specific mechanisms associated with pre-eclampsia complicated with growth restriction, or may simply reflect the presence of syncytiotrophoblast layer damage, regardless of underlying pathological condition.
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Affiliation(s)
- Rigmor Austgulen
- Faculty of Medicine, Institute of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, 7489 Trondheim, Norway.
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Yamauchi H, Katayama KI, Ueno M, Uetsuka K, Nakayama H, Doi K. Involvement of p53 in 1-beta-D-arabinofuranosylcytosine-induced trophoblastic cell apoptosis and impaired proliferation in rat placenta. Biol Reprod 2004; 70:1762-7. [PMID: 14766721 DOI: 10.1095/biolreprod.103.026252] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/01/2022] Open
Abstract
1-beta-D-Arabinofuranosylcytosine (Ara-C), a DNA-damaging agent, severely inhibits fetal growth and has teratogenicity. Recently, we reported that Ara-C also causes placental growth retardation and increases placental apoptosis. The aim of the present study is to elucidate the mechanisms of placental injury induced by genotoxic stress and involvement of p53, which mediates apoptosis and cell-cycle arrest after DNA damage. We injected Ara-C into pregnant rats on Day 13 of gestation and examined the placentas from 1 to 48 h after the administration. Terminal deoxynucleotidyltransferase-mediated dUTP end-labeling (TUNEL) revealed that the apoptosis of trophoblastic cells in the placental labyrinth zone increased from 3 h after the treatment and peaked at 6 h before returning to control levels at 48 h. An increase in cleaved caspase-3 immunoreactivity was also detected at 6 h. Proliferative activity as measured by immunohistochemistry for topoisomerase II alpha and by mitotic index significantly decreased after the treatment in the labyrinth zone. Immunoreactivity for p53 protein in the placental labyrinth zone was remarkably enhanced and peaked at 3 h after treatment, although no increase in p53 mRNA expression was detected with a reverse transcription- polymerase chain reaction. Regarding p53 target genes, p21, cyclinG1, and fas mRNA levels increased significantly and peaked at around 9 h after the treatment. These results indicate that Ara-C would induce apoptosis and impair cell proliferation in the placental labyrinth zone, and p53 and its transcriptional target genes may play an important role in the pathogenesis of the Ara-C-induced placental toxicity.
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Affiliation(s)
- Hirofumi Yamauchi
- Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo,Tokyo 113-8657, Japan
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Wang X, Yi S, Athayde N, Trudinger B. Endothelial cell apoptosis is induced by fetal plasma from pregnancy with umbilical placental vascular disease. Am J Obstet Gynecol 2002; 186:557-63. [PMID: 11904623 DOI: 10.1067/mob.2002.121257] [Citation(s) in RCA: 18] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
OBJECTIVE Vascular disease in the umbilical placental circulation that is detected by umbilical artery Doppler study is associated with adverse fetal outcome. Endothelial cell activation and platelet consumption are features of this pathologic condition. We postulated that this was due to the local release of factors that cause endothelial cell injury and that these would spill into the fetal circulation. To test this hypothesis, we examined for the presence in fetal plasma of factors that induced endothelial cell apoptosis in pregnancies that were complicated by umbilical placental vascular disease. STUDY DESIGN Isolated and cultured human umbilical vein endothelial cells were exposed to fetal plasma from the 3 fetal groups: normal pregnancy (n = 32 patients), pregnancy with umbilical placental vascular disease that was identified by an abnormal umbilical artery Doppler study (n = 38 patients), and pregnancy with maternal preeclampsia and normal umbilical artery Doppler study (n = 16 patients). Early apoptosis can be recognized by a loss of plasma membrane asymmetry with membrane uptake of annexin V. This was measured with annexin V and propidium iodide staining by fluorescent-activated cell scanning. Cells that underwent early apoptosis stained positive for annexin V and negative for propidium iodide (in contrast with cells that underwent necrosis). Cytosolic proteolytic activity was also measured. The lysates from endothelial cells that were stimulated by fetal plasma from umbilical placental vascular disease were tested for caspase-3 and caspase-8 activities by a fluorescent assay with spectrofluorophotometry. RESULTS The percentage of endothelial cells that underwent apoptosis was significantly higher (P <.05) when stimulated with fetal plasma from pregnancies with umbilical placental vascular disease (17.71% +/- 1.31%) than with fetal plasma from normal pregnancies (9.76% +/- 0.87%). In the presence of maternal preeclampsia with normal umbilical artery Doppler study, the percent of apoptotic cells (11.31% +/- 1.59%) was similar to that of the normal group. In the group with abnormal umbilical artery Doppler study, there was no difference between pregnancies with preeclampsia (n = 17 pregnancies) and without preeclampsia (n = 21 pregnancies). The protease activity of caspase-3 was significantly enhanced in the group with umbilical placental vascular disease compared with normal pregnancy (0.79 +/- 0.06 vs 0.45 +/- 0.08 microMol/L). However, no difference in caspase-8 activity was detected (0.66 +/- 0.05 vs 0.56 +/- 0.05 microMol/L). CONCLUSION Endothelial cell apoptosis is a feature of umbilical placental vascular disease. Our study demonstrates the presence of factors in the fetal plasma that caused endothelial cells to undergo early apoptosis. This increased apoptosis was only seen in the presence of placental vascular disease and was independent of the presence or absence of maternal preeclampsia. Our results indicate that programmed endothelial cell death occurs in the fetal circulation as a part of the injury that is associated with the development of umbilical placental vascular disease. The caspase-3, rather than caspase-8, signal transduction pathway appears to be involved in the mediation of endothelial cell apoptosis that was detected in our study.
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Affiliation(s)
- Xin Wang
- Department of Obstetrics and Gynaecology, University of Sydney at Westmead Hospital, NSW, Australia
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