AMH regulates ovarian granulosa cell growth in PCOS rats through SMAD4

Abstract

Abstract Background Polycystic Povary syndrome(PCOS) is a diverse condition with an unknown cause. Anti-Mullerian hormone(AMH) is a hormone that belongs to the transforming growth factor-β(TGF-β) class. Mothers against decapentaplegic homolog 4(SMAD4) is a crucial transcription factor widely expressed in granulosa cells in the TGF-β signaling pathway. Previous studies have revealed that AMH may be an important factor in follicular developmental disorders in PCOS patients , as a biomarker of PCOS. Objective This study examines the involvement of AMH in the formation of ovulatory abnormalities in PCOS rats and explores its potential causes. Methods A PCOS rat model was created by inducing DHEA, and granulosa cells from the ovaries were extracted and identified. The expression of AMH and SMAD4 in PCOS rats was assessed by ELISA, immu-nohistochemistry, and Western blot; and the effects of different concentrations of AMH recombinant proteins on the expression of SMAD4 and the development of granulosa cells were examined. The effect of knocking down SMAD4 expression with siRNA on granulosa cell development was also examined. Results The expression of AMH and SMAD4 in the ovarian tissues and granulosa cells in the PCOS group was higher (*P<0.05). The expression of PCNA in the ovarian granulosa cells of the PCOS group was lower (**P<0.01), and the expression of BAX was higher (*P<0.05). Western blot analysis indicated that 100 ng/ml rAMH increased the expression of SMAD4 and caspase-3 in granulosa cells (*P<0.05), and decreased CyclinA and BCL-2 expression (**P<0.01). CCK-8 and flow cytometry results showed that 100 ng/ml AMH reduced proliferation and increased apoptosis in granulosa cells (***P<0.001,*P<0.05). siRNA knockdown of the SMAD4 gene led to increased PCNA expression (**P<0.01) and decreased BAX expression (*P<0.05) in granulosa cells of PCOS rats. Conclusions AMH plays a role in controlling the growth and programmed cell death of ovarian granulosa cells in rats with polycystic ovary syndrome through SMAD4.