Human Placenta-Derived Mesenchymal Stem Cells Transplantation Can Restore Ovarian Function In Autoimmune-Induced Premature Ovarian Failure Mice

1.Restoring ovarian function with hPMSCs transplantation into autoimmune-induced POF mice mediated by Treg cells and TGF-β,IFN-y cytokinesObjective:To explore the restoring function with hPMSCs transplantation into autoimmune-induced POF Mice,and to investigate its association with Treg cells and associated cytokines(TGF-β and IFN-y).Methods:Following ZP3 treatment,hPMSCs suspension were intravenously injected into part of mice,which were chosen with the qualitative detection of AZPAb by ELISA.Estrous cycle,ovarian morphologic changes and folliculogenesis were measured by HE detection.The serum levels of E2 were detected by ELISA,as well as the FSH detection.And the detection of apoptotic GCs in ovarian tissues were performed by IHC.To determine if immune response is involved in the regulation of ovarian function change,the population of Treg cell populations were detected by FCM and expression of associated cytokines,e.g.,TGF-β and IFN-y were measured by ELISAResults:After hPMSCs transplantation,the injured ovarian function is significantly improved.Compared with mice only receiving ZP3 treatment,the estrous cycles were almost back to normal in mice receiving hPMSCs transplantation,and the degree of ovarian fibrosis were reduced,also,with increased functional follicles and decreased atretic follicles detected Serum levels of E2 and FSH were nearly the same with those in normal mice.Additionally,significantly decreased apoptotic GCs were also detected.The transplantation of hPMSCs significantly increased the population of Treg cell which were inhibited by ZP3 treatment,and the serum levels of decrease in TGF-β and increase in IFN-γ caused by ZP3 treatment have been reversed following hPMSCs transplantationConclusion:HPMSCs transplantation can restore ovarian function into POF mice,and the function may through up-regulating the expression of Treg cells and TGF-β,and down-regulating the expression of IFN-y.2.HPMSCs transplantation restoring ovarian function in premature ovarian failure mice may associated with the change of Th17/Tc17 and Th17/Treg cells ratio through activating PI3K/Akt signal pathwayObjective:To investigate whether PI3K/Akt signal pathway could be involved in the ovarian restoring function with hPMSCs transplanted into autoimmune-induced POF mice,and whether the restoring function could be associated with Th17,Tc17 and Treg cells.Methods:HPMSCs were intravenously injected into mice following ZP3 treatment in vivo.LY294002,the inhibitor of PI3K/Akt,were administrated two weeks after hPMSCs transplantation.Serum levels of E2,FSH,LH,AMH and AZPAb were detected by ELISA,the ovarian morphological changes were observed by HE staining,and the expression of caspase-3 in the ovarian tissue detected by IHC were used to observe the apoptosis of GCs,the expression of Akt and p-Akt protein were detected by IHC and WB assay in vivo.The ovarian GCs were separated and cultured with or without the supernatant produced by hPMSCs,and observed under the influence of different concentrations of LY294002.To further explore the relationship of the restoring function with immune factors,the percentage of Th 17,Tc 17 and Treg cells were detected by FCM,along with the expression of interleukin(IL)-17 in blood sample measured by ELISA.Also,a MTT assay to assess cell proliferation and a FCM to analyze cell apoptosis were performed in vitro.Results:With LY294002 administration,serum levels of E2 and AMH were down-regulated,while serum levels of FSH,LH and AZPAb showed up-regulated compared with mice receiving hPMSCs transplantation.The ovarian morphology presented as atrophy and fibrosis with functional follicles exhausted,also,GCs apoptosis shown significantly increased.The expression of p-Akt in the ovary showed decrease.The ratios of Th17/Tc17 and Th17/Treg,as well as the serum levels of IL-17,were significantly increased with LY294002 administrated.Thus,LY294002 significantly decreased GCs proliferation and markedly increased apoptosis of GCs.Conclusion:Recovering ovarian function in POF mice with hPMSCs transplantation may associate with activated PI3K/Akt signal pathway through down-regulating the ratios of Th17/Tc17 and Th17/Treg cells.