Affects And Mechanisms Of ESCs-sEVs On POI

ObjectiveThe objective of this study is to evaluate the curative effect of small extracellular vesicles derived from embryonic stem cells on premature ovarian failure mice and further explore the possible molecular mechanism implicated in its actions.MethodIn this study,a mouse model of premature ovarian insufficient(POI)was established by intraperitoneal injection of the chemotherapy drugs cyclophosphamide and busulan.Small extracellular vesicles derived from embryonic stem cells(ESCs-sEVs)were administered via tail vein injection.The serum estradiol,follicle stimulating hormone,and anti-Mullerian hormone contents of the mice were measured at 1 and 2 weeks after treatment,respectively.HE staining was used to evaluate ovarian structure and number of follicles,and TUNEL staining was used to observe apoptosis of mouse ovarian granulosa cells.Then the mouse ovarian granulosa cells were extracted and cultured in vitro.After the intervention of ESCs-sEVs,the proliferation ability of granulosa cells was detected by EdU and CCK-8 experiments.Western-blot was used to detect PI3K/AKT pathway and downstream apoptosis-related protein expression.ResultsAfter treatment with ESCs-sEVs,serum sex hormones of mice returned to normal levels,and the number of follicles at various levels in the ovary increased,and the number of apoptotic granulosa cells decreased.Further research found that after ESCs-sEVs intervention,the proliferation of granulosa cells increased,PI3K/AKT pathway was simultaneously activated,the expression of downstream apoptosis proteins Caspase-3 and Bax decreased,and the expression of anti-apoptotic protein Bcl-2 increased.Moreover,when the PI3K/AKT pathway is inhibited,the affects of ESCs-sEVs on the proliferation and anti-apoptosis of granulocytes are weakened.ConclusionThis study found that ESCs-sEVs can activate the PI3K/AKT pathway,affect the activity of ovarian granular cells and promote the recovery of ovarian function in mice,providing new ideas and directions for the clinical treatment of premature ovarian insufficient.