BMP15 Regulates AMH Expression By P38 MAPK Passway In The Goat Ovary

Individual AMH level is directly related with ovarian reserve.The higher level of AMH means the stronger ovarian reserve in individual.It has been confirmed that AMH level can be used as the evaluation criteria of ovarian reserve.On the regulation of AMH expression,domestic and foreign research has confirmed that BMPs family plays a role of regulating the expression of AMH by SMAD pathway,and literatures have been reported that BMP15 can up-regulate the expression of AMH in human and sheep ovary.But the specific mechanism of BMP15 regulating AMH is not clear so far.Chongqing local goats in sexual maturity were used as experimental samples in order to analyze the mechanism of BMP15 regulating AMH expression.And both sides of the ovaries were collected to culture granulosa cells.The expression of AMH in granulosa cells was up-regulated and p38 MAPK phosphorylation was activated by 10 ng/mL of BMP15 after different concentrations of exogenous recombinant human(rh)BMP15 protein dealt with granulosa cells.On this basis,the granulosa cells were dealt with 10 ng/m L of BMP15 with or without p38 MAPK pathway inhibitor SB203580 or with or without siRNA of p38 MAPK,and the expression of AMH and AMH transcription factor related gene SOX9 were measured in both transcriptional level and translational level.According to the expression changes of each factor,the role of p38 MAPK pathway in the process of BMP15 regulating AMH was speculated.The main results were as follows:1.The expression of AMH was up-regulated by BMP15 in granulosa cells of goats,and the 100 ng/mL of BMP15 had more significant effect on increasing the expression of AMH.At the same time,the phosphorylation of p38 MAPK was activated by the concentration of 10 ng/mL of BMP15.2.The concentration of the inhibitor under 25 μM would not affect the activity of granulusa cells,and when the inhibitor concentration was 20μM,the effect was best on the p38 MAPK passway.Compared with the cells dealt with BMP15,the granulusa cells that dealt with BMP15 and SB203580 had a lower AMH expression and SOX9 expression(P<0.05),what suggested that BMP15 up-regulated AMH and SOX9 expression by p38 MAPK passway.3.The expression of p38 MAPK gene could be silenced by interference fragment of p38 MAPK.The expression of AMH and SOX9 were significantly decreased(P<0.05)after the granulosa cells dealt with BMP15 after p38 MAPK gene silenced,which further proved that p38 MAPK pathway plays an important role in the process of BMP15 regulating AMH.The results of this research indicated that BMP15 stimulate the expression of AMH mainly by activating p38 MAPK pathway.SOX9 is involved in the process.This study provided basic data for the research of AMH,and also laid a solid foundation to clear the molecular mechanism of regulation of AMH expression.