Effect Of CYP19A1 In Rat Ovary With Polycystic Ovary Syndrome By Human Menstrual Blood-derived Mesenchymal Stem Cells

Background:Polycystic ovary syndrome?PCOS?is a common endocrine disease in women of childbearing age that can lead to infertility.The increase in androgen in PCOS?Polycystic ovary syndrome?patients is closely related to a series of pathological changes.Aromatase,encoded by CYP19A1,can catalyze the transformation of androgen to estrogen.Mesenchymal stem cells?MSCs?are currently used in the treatment of many diseases,among which the human menstrual blood-derived mesenchymal stem cells?MenSCs?are newly found,and there is no research report for the treatment of polycystic ovary syndrome.Objective:This study investigated the relationship between human menstrual blood-derived mesenchymal stem cells?MenSC?therapy and CYP19A1 mRNA expression of ovaries in rats with PCOS.Methods:Sixty 6-week-old female rats were randomly selected,fourty rats were injected with insulin+hCG,and twenty rats were set as the control group.The changes of estrous cycle were observed by detecting the concentration of serum testosterone and the changes of vaginal exfoliated cells.Four rats in PCOS group and two rats in control group were sacrificed,ovaries were taken,and one side of ovaries was sectioned to observe pathological changes,the other ovary was used for RNA extraction and protein extraction to detect the success of polycystic ovary modeling.Human menstrual blood-derived mesenchymal stem cells?MenSC?were cultured,and their surface antigen molecules were identified by flow cytometry,and the differentiation function of the cells was identified;Some of the cells were transfected with lentivirus with genes carrying Luciferase Report System and green fluorescent protein gene.After culture and proliferation,they were injected into rats through tail vein to observe the location of their implantation and whether there was any implantation in ovaries.There were 15 rats in PCOS-MenSCs group,9 rats in PCOS-DMEM group,9 rats in NC-MenSCs group and 8rats in NC-DMEM group.The estrous cycle of rats in each group was dynamically observed.Eighteen days after injection,blood samples were collected to observe changes in serum testosterone;We sacrificed the rats and took bilateral ovarian tissues.One side of the ovaries were paraffin sections to observe the pathological changes.The other side of the ovary was used to extract RNA and protein to be tested.RNA was extracted from the ovaries of each group of rats,and the RNA was reverse transcribed into cDNA by PCR,The concentration was detected by real-time quantitative PCR.After obtaining the C?t?value,double-tailed t-test analysis was performed to analyze the increase in CYP19A1expression in PCOS rats after MenSC treatment(Take 2^-?C?t?value as the comparison standard.).And we calculated the 2^-??C?t?value to get the difference change value.Results:The estrous cycle of PCOS model group was disordered,and serum testosterone levels of PCOS model rats was significantly higher than that of the control group,the differences were statistically significant.Identification of menstrual blood-derived mesenchymal stem cells:We used flow cytometry to identify cell surface molecules.The results showed that CD34/?-?,CD45/?-?,HLA-DR/?-?,CD44/?+?,CD73/?+?,CD90/?+?,CD105/?+?;After corresponding stimulation,mesenchymal stem cells successfully induced adipogenic differentiation,chondrogenic differentiation and osteogenic differentiation,these results are in line with the international standards for the identification of mesenchymal stem cells.CYP19A1 mRNA expression of PCOS rats in the MenSC-treated group was significantly higher than that of PCOS rats in the DMEM-treated group,control rats in the MenSC-treated group,and PCOS rats in the DMEM-treated group?P<0.05?.The serum testosterone levels of PCOS rats treated with MenSCs were lower than that of untreatedPCOS rats?P<0.05?.Conclusions:The cells isolated from human menstrual blood by relevant laboratory researchers had been proved to be MenSCs in our study;Subcutaneous injection of insulin+hCG can successfully induce PCOS rat model.Menscs can successfully homed into ovary by tail vein transplantation in mice.MenSCs implanted into the ovaries of rats can promote CYP19A1 expression and reduce serum testosterone concentration,thus having significance for PCOS treatment.