Mechanism Of MiR-199a-3p Promoting Apoptosis Of PCOS Granulosa Cells

Objective: Polycystic ovary syndrome(PCOS)is the most common reproductive and metabolic disorder in women of childbearing age.The expression of miR-199a-3p in follicular fluid of PCOS patients is increased,while its mechanism is not clear.The purpose of this paper is to study the effect of miR-199a-3p on glycolysis and apoptosis of PCOS-like granulosa cells at cellular level,in order to explore the mechanism of miR-199a-3p in the pathogenesis of PCOS at the molecular level,so as to provide a new experimental basis for further expounding the etiology of PCOS.Methods: QRT-PCR was used to detect the expression of miR-199a-3p in follicular fluid of PCOS patients and normal women.KGN cell line was cultured in vitro,and the PCOS granulosa cell model was constructed by testosterone intervention.The expression of miR-199a-3p in PCOS granulosa cells was detected by qRT-PCR.Determination the glycolytic metabolites of granulosa cells after testosterone intervention by ELISA;Bioinformatics analysis verified the potential targeting relationship between miR-199a-3p and LDHA.After transfection of mir-199a-3p inhibitors and analogues,qRT-PCR was used to detect the expression of mir-199a-3p、 LDHA、 proliferation and apoptosis related genes.Cell viability was measured by CCK8,and apoptosis was measured by flow cytometry.Results: 1.QRT-PCR showed that the expression of miR-199a-3p in follicular fluid of PCOS patients was significantly higher than that of normal women.2.There was miR-199a-3p expression in KGN cells,and the expression of miR-199a-3p increased after testosterone intervention.3.Testosterone promotes the production of pyruvate and inhibits the production of lactic acid.4.Bioinformatics analysis showed that miR-199a-3p could bind to LDHA.5.Mi R-199a-3p increased pyruvate production,decreased lactate production and decreased LDHA expression.6.Mi R-199a-3p decreased the activity and increased the apoptosis of granulosa cells.Conclusions: MiR-199a-3p can promote the apoptosis of PCOS granulosa cells,and its mechanism may be related to inhibiting the expression of LDHA in the glycolytic pathway of granulosa cells and affecting the energy metabolism of granulosa cells.