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Bisphenol A promotes autophagy in ovarian granulosa cells by inducing AMPK/mTOR/ULK1 signalling pathway
Highlights Higher BPA exposure may influence the early outcomes of IVF-ET. BPA could induce autophagy and endocrine dysfunction in GCs. AMPK/mTOR/ULK1 pathway might be the key player in BPA-induced autophagy in GCs.
Abstract Background Bisphenol A (BPA) is a widespread endocrine-disrupting chemical with estrogen like effects, which could interfere with the human reproductive system by disrupting the normal function of granulosa cells (GCs) leading to abnormal ovarian function. However, the mechanism of its toxicity on human GCs has not been clearly described thus far. Methods 106 normogonadotropic infertile women undergoing their first in-vitro fertilization-embryo transfer (IVF-ET) cycle were recruited. Urinary BPA level and the early outcomes of IVF-ET were analysed. Patients were divided to low and high BPA exposure groups using the median urinary BPA concentration as the cut-off value. In-vivo and in-vitro studies were conducted using mice and human granulosa cell line (KGN cells). Female Kunming mice approximately 6–8 weeks of age were poisoned with BPA at different dosages (1, 10 or 100 μg/kg) by oral gavage once daily for 2 weeks, while KGN cells were exposed to BPA at the concentration of 1, 10 or 100 nM for 24 h, 48 h or 72 h. BPA-induced ovarian morphologic changes were analysed by histopathology investigation. Cell viability and apoptosis were evaluated using CCK-8, TUNEL and flowcytometric, respectively. Hormone levels were determined using ELISA and the molecular mechanism studies were conducted using immunofluorescence, RT-PCR and western blots. Results The oocyte retrieval rate, maturation rate and embryo implantation rate significantly decreased with the higher level of urinary BPA concentration. Peak E2 level was lower in high BPA group, but no statistical significance could be observed. In BPA treated mice, cystic dilation of the follicles with a decreased number of GCs could be observed histopathologically. Decreased E2, P4 and AMH level and GCs autophagy could be detected both in-vivo and in-vitro with the activation of AMPK/mTOR/ULK1 signalling pathway. As being confirmed in KGN cells, phosphorylated AMPK and ULK1 increased while phosphorylated mTOR decreased, and by inhibition autophagy using knockdown of AMPK or 3-MA, adverse effects of BPA exposure in-vitro could be reversed. Conclusion BPA exposure might abnormally influence human ovarian functions leading to abnormal folliculogenesis by activation of autophagy in GCs through AMPK/mTOR/ULK1 pathway.
Bisphenol A promotes autophagy in ovarian granulosa cells by inducing AMPK/mTOR/ULK1 signalling pathway
Highlights Higher BPA exposure may influence the early outcomes of IVF-ET. BPA could induce autophagy and endocrine dysfunction in GCs. AMPK/mTOR/ULK1 pathway might be the key player in BPA-induced autophagy in GCs.
Abstract Background Bisphenol A (BPA) is a widespread endocrine-disrupting chemical with estrogen like effects, which could interfere with the human reproductive system by disrupting the normal function of granulosa cells (GCs) leading to abnormal ovarian function. However, the mechanism of its toxicity on human GCs has not been clearly described thus far. Methods 106 normogonadotropic infertile women undergoing their first in-vitro fertilization-embryo transfer (IVF-ET) cycle were recruited. Urinary BPA level and the early outcomes of IVF-ET were analysed. Patients were divided to low and high BPA exposure groups using the median urinary BPA concentration as the cut-off value. In-vivo and in-vitro studies were conducted using mice and human granulosa cell line (KGN cells). Female Kunming mice approximately 6–8 weeks of age were poisoned with BPA at different dosages (1, 10 or 100 μg/kg) by oral gavage once daily for 2 weeks, while KGN cells were exposed to BPA at the concentration of 1, 10 or 100 nM for 24 h, 48 h or 72 h. BPA-induced ovarian morphologic changes were analysed by histopathology investigation. Cell viability and apoptosis were evaluated using CCK-8, TUNEL and flowcytometric, respectively. Hormone levels were determined using ELISA and the molecular mechanism studies were conducted using immunofluorescence, RT-PCR and western blots. Results The oocyte retrieval rate, maturation rate and embryo implantation rate significantly decreased with the higher level of urinary BPA concentration. Peak E2 level was lower in high BPA group, but no statistical significance could be observed. In BPA treated mice, cystic dilation of the follicles with a decreased number of GCs could be observed histopathologically. Decreased E2, P4 and AMH level and GCs autophagy could be detected both in-vivo and in-vitro with the activation of AMPK/mTOR/ULK1 signalling pathway. As being confirmed in KGN cells, phosphorylated AMPK and ULK1 increased while phosphorylated mTOR decreased, and by inhibition autophagy using knockdown of AMPK or 3-MA, adverse effects of BPA exposure in-vitro could be reversed. Conclusion BPA exposure might abnormally influence human ovarian functions leading to abnormal folliculogenesis by activation of autophagy in GCs through AMPK/mTOR/ULK1 pathway.
Bisphenol A promotes autophagy in ovarian granulosa cells by inducing AMPK/mTOR/ULK1 signalling pathway
Lin, Miaoling (Autor:in) / Hua, Rui (Autor:in) / Ma, Jing (Autor:in) / Zhou, Yao (Autor:in) / Li, Pei (Autor:in) / Xu, Xiya (Autor:in) / Yu, Zhiqiang (Autor:in) / Quan, Song (Autor:in)
20.11.2020
Aufsatz (Zeitschrift)
Elektronische Ressource
Englisch
Bisphenol A , Sex hormones , Autophagy , Apoptosis , AMPK , BPA , bisphenol A , IVF/ICSI , in vitro fertilization/intracytoplasmic sperm injection , GC , granulosa cell , RAPA , rapamycin , 3-MA , 3-methyladenine , shRNA , short hairpin RNA , LDH , lactate dehydrogenase , LH , luteinizing hormone , AMH , anti-Mullerian hormone , E2 , oestradiol , P4 , progesterone , adenosine 5′-monophosphate (AMP)-activated protein kinase , ULK1 , unc-51-like kinase 1 , mTOR , mammalian target of rapamycin , PCOS , polycystic ovary syndrome , POF , premature ovarian failure , GCT , granulosa cell tumours , RT-PCR , real-time PCR , CCK-8 , Cell Counting Kit-8 , IF , immunofluorescence staining , GnRH-a , gonadotropin-releasing hormone agonist , shRNA-AMPK , shRNA against AMPK , NC , negative control , TUNEL , TdT-mediated dUTP nick-end labelling , ELISA , enzyme-linked immunosorbent assay , ROS , reactive oxygen species , FBS , foetal bovine serum , DMEM , Dulbecco's modified Eagle’s medium
| DOAJ | 2021