The Mechanism Of Mouse Ovarian Follicle Development Regulated By AMPK-mTOR-HIF1-VEGF Pathway

Aim: To determine the regulation mechanism of mouse follicle development through AMPK-mTOR-HIF1-VEGF pathway by using AMPK inhibitor,compound C.Method: The changes in AMPK-mTOR-HIF1-VEGF pathway,follicle development or blood vessel growth of the ovaries from wild or Vegf+/? heterozygote,10 d or 3w mice,in vitro cultured,in vivo transplanted or intrabursal injected with compound C or DMSO which was as control,were detected by H&E staining,CLARITY technique with immunohistochemistry,Western blotting or quantitative rt-PCR.Result: The ovarian phosphorylation of mTOR and downstream proteins,including eIF4 B and S6,in AMPK-mTOR-HIF1-VEGF signaling pathway was upregulated by compound C while TSC was downregulated.And compound C also increased the expression of HIF-1α and VEGF gene,activated primordial follicle in vitro,promoted primary,early and late secondary follicle growth in vitro,developed primary,antrum follicle and blood vessel growth in vivo.Intrabursal injection with compound C combined with controlled ovarian hyperstimulation can increase the number of ovulated oocytes in wild type mice,but not Vegf+/? hetrozygote mice.Conclusion: AMPK inhibitor can activate primordial follicle,promote follicle development and blood vessel growth through AMPK-mTOR-HIF1-VEGF pathway.This study may deepen our understanding of intraovarian mechanisms underlying early folliculogenesis,help to find out the hidden pathogenesis in the patients diagnosed with primary ovarian insufficiency,polycystic ovary syndrome,and poor ovarian response to FSH stimulation,as well as for infertile women of advanced reproductive age,and allow us to develop novel therapeutic strategies for them.