1. Vigilin interacts with ER-β to protect against palmitic acid-induced granulosa cells apoptosis via inhibiting calcineurin-mediated Drp1 signaling pathway
- Author
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Yu-Lin Nie, Jun Zhou, Yang Cao, Mei-Qing Li, Liao Hongqing, and Jing Zhou
- Subjects
Mitochondrial ROS ,Dynamins ,Clinical Biochemistry ,Palmitic Acid ,030209 endocrinology & metabolism ,Apoptosis ,Mitochondrion ,Biochemistry ,Flow cytometry ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Western blot ,Cell Line, Tumor ,medicine ,Estrogen Receptor beta ,Humans ,Viability assay ,RNA, Messenger ,Molecular Biology ,Pharmacology ,Granulosa Cells ,medicine.diagnostic_test ,Chemistry ,Calcineurin ,Organic Chemistry ,RNA-Binding Proteins ,Cell biology ,Gene Expression Regulation ,Cell culture ,Cytoprotection ,030220 oncology & carcinogenesis ,Female ,Signal transduction ,Signal Transduction - Abstract
Background Hypercholesterolemia is one of the causes of female infertility, and as a common fatty acid in follicular fluid, palmitic acid (PA) level plays a vital role in granule cell which is closely related to the developmental potential of follicle. Methods The ovarian granulosa cell-like human granulosa (KGN) cell line and the immortalized normal ovarian surface epithelial cell line (IOSE80) were used to verify the effect of PA on cell viability and apoptosis by MTT and flow cytometry assay, respectively. Then mitochondria damage was confirmed by mitochondrial membrane potential, mitochondrial ROS detection assay and western blot in KGN cells. Thorough luciferase reporter assay and RIP-qPCR, the relationship between vigilin and ER-β was investigated. Results In our study, PA induced mitochondrial damage-mediated cell apoptosis of KGN cells was dose-dependently, while PA shown no effects on in IOSE80 cells. Then the role of calcineurin (CnA)-mediated Drp1 signaling pathway on KGN cells was confirmed by treating with Mdivi-1 or FK506T. In addition, the changed level of vigilin and ER-β was observed in cell apoptosis of KGN cells induced by PA. By transfecting vigilin vector or ER-β vector into KGN cells, respectively, vigilin and ER-β were demonstrated to regulate the apoptosis of KGN cells. And vigilin was a binding protein of ER-β mRNA. Conclusion Vigilin could interact with ER-β mRNA to promote ER-β expression. And Vigilin/ ER-β relieve the mitochondrial damage and cell apoptosis induced by PA through regulating CnA-mediated Drp1 signaling pathway, which revealed the mechanism and strategy of hypercholesterolemia in female infertility.
- Published
- 2020