Effect And Mechanism Of Adipose-derived Mesenchymal Stem Cells In The Treatment Of Chemotherapy-induced Premature Ovarian Failure

Premature ovarian failure is the phenomenon of premature ovarian function decline.The specific clinical feature are markedly elevated gonadotropin(more than40IU/L),markedly reduced estrogen,and amenorrhea at least four years before the age of 40.There are many causes of premature ovarian failure,one of them is due to chemotherapeutics,which is called chemotherapy-induced premature ovarian failure(CIPOF).Premature ovarian failure seriously affects women’s health,including fertility,bone mineral density,cardiovascular,nervous system,mental health,etc.Therefore,as long as it does not interfere with cancer treatment,treatment of chemotherapy-induced premature ovarian failure is an urgent clinical problem.With the rapid development of regenerative medicine,stem cells have played a huge role in the treatment of various diseases.In recent years,adipose-derived mesenchymal stem cells have broad prospects of application in the field of regenerative medicine.Adipose-derived mesenchymal stem cells can secrete a series of functional factors,participating in the repair of angiogenesis,immune regulation,and inflammation.Therefore,adipose-derived mesenchymal stem cells are expected to become a new method for the treatment of premature ovarian failure.Based on the analysis of proteomics and bioinformatics,the effect and mechanism of adipose-derived Mesenchymal Stem Cells in the treatment of premature ovarian failure were studied to provide theoretical support for clinical treatment.Methods:Inguinal fat of SD rats was isolated,primary cultured,cell phenotype was identified by flow cytometry,osteogenic and adipogenic ability of differentiation induction was detected,and cell morphology was observed,providing a basis for subsequent cell transplantation.The following experiments were employed 3-5generations of mesenchymal stem cells.First,an animal model of chemotherapyinduced ovarian insufficiency was established by intraperitoneal injection of cisplatin.Seven days after the modeling,the rats were randomly divided into three groups,namely the control group(PBS group)and the model group(cisplatin group).,treatment group(transplanted ADSCs group).ADSCs were transplanted by intraperitoneal injection for treatment.In the experiment,the general state of the rats was observed,the body weight was weighed.after ADSCs transplantation for one week,the ovarian tissue frozen section was stained with HE to evaluate the morphological structure of the ovarian tissue.During the experiment,the estrous cycle of the rats was monitored,and the serum levels of FSH,LH,and E2 were detected by ELISA to explore the repair of adipose-derived mesenchymal stem cells to the damage of ovarian function in rats.The whole protein was extracted from ovarian tissue,and the differentially expressed genes between the treatment group and the model group were analyzed by bioinformatics,and the differentially expressed genes were subjected to KEGG,Mfuzz cluster analysis,which analysis the functions of differentially expressed genes,related pathways and the interaction of proteins.Based on proteomics to explore the role of transplantation of ADSCs in the treatment of chemotherapy-induced premature ovarian failure,providing relevant theoretical basis for the treatment of chemotherapy-induced ovarian insufficiency.Result:(1)After the third generation,adipose-derived mesenchymal stem cells are dendritic fibroblast-like cells.Adipose-derived mesenchymal stem cells have multidirectional differentiation potential,highly expressing stem cell marker CD90,low expressing non-stem cell markers CD44,CD45,CD103.(2)The estrous cycle of the rats in the model group was disordered,and the specific manifestations were prolonged interval,rough hair,decreased activity,and body weight was significantly lower than the control group.After two weeks of treatment,The cycle of the rats in the treatment group gradually returned to normal and the general condition improved.(3)The ovarian structure of the model group was obviously disordered,normal follicles were not found,and atretic follicles increased.After transplantation of adipose-derived mesenchymal stem cells,the number of follicles in the ovary increased,the number of atretic follicles decreased,and the degree of interstitial fibrosis was alleviated.(4)Compared with the control group,the serum E2 level in the model group was significantly lower(P<0.05),and the FSH and LH level were increased compared with the control group(P<0.05).The serum levels of FSH and LH in the treatment group were decreased,and the E2 level was increased(P<0.05).(5)The differentially expressed proteins were screened.Compared with the control group,the model group had 119 up-regulated proteins and 62 down-regulated proteins.Compared with the model group,the transplanted ADSCs group had 77 upregulated proteins and 81 down-regulated proteins.The protein expression pattern was analyzed,and the proteins whose expression changed before and after treatment were screened out.The pathway enrichment analysis was performed to screen out typical pathways.The results showed that the levels of PI3 K and TIMP-1 were decreased,and the levels of LKB-1 were up-regulated.Conclusion:(1)ADSCs can improve the endocrine dysfunction of the ovary caused by chemotherapy,improve the ovarian structure damage in model mice,and have a certain effect in the treatment of premature ovarian failure.(2)ADSCs in the treatment of premature ovarian failure mainly play a role in PI3 k,HIF-1/VEGF,LKB-1/AMPK and other pathways by reducing the levels of TIMP-1 and PI3 K and increasing the level of LKB-1.