Effect Of Metformin On Endometrial Receptivity In Patients With PCOS By MiR-1910-3p And MiR-491-3p

Polycystic ovarian syndrome(PCOS)is a common disease affecting female reproductive endocrine disorders in the growth period.It is mainly manifested as rare ovulation or anovulation,clinical or biochemical hyperandrogenism and imaging ovarian polycystic changes.Some with insulin resistance(IR),obesity and seriously affect the fertility of women of childbearing age.The prerequisite for successful pregnancy is the successful implantation of the embryo into the endometrium,while requiring high quality embryos and well tolerated endometrium,as well as complex interactions between the two.Endometrial receptivity(ER)refers to the ability of endometrial receptive embryos,the human endometrium in the menstrual cycle on the 19 th to 23 days to accept the embryo,this period called the window of implantation(WOI).There is a good signal pathway in the endometrium with good receptivity,which promotes the interaction between the embryonic cells and the endometrial stroma.Many studies have shown that PCOS endometrial tolerance affected embryo implantation,abortion rate increased.Because PCOS patients with insulin resistance,and metformin as the most commonly used clinical insulin sensitizer,also used in PCOS patients.Metformin can improve the sensitivity of cells to insulin and improve the metabolism of tissues and organs.It has been reported that metformin can improve the pregnancy rate ofPCOS patients by improving the receptivity of endometrium and reduce the abortion rate,but the mechanism is unclear.Endometrial receptivity is regulated by a variety of molecules and genes,including microRNAs(miRNAs).MiRNAs are a class of non-coding single-stranded RNA molecules that regulate mRNA and protein expression at both transcribed and post-transcriptional levels by completely or non-completely complementary to the 3'untranslated region(3'-UTR)of the target gene.MiRNAs are involved in the establishment of endometrial receptivity and the process of embryo implantation.Previous studies have shown that the rate of pregnancy after treatment with PCOS infertility is significantly increased,abortion rates are significantly reduced,endometrial receptive molecular markers HOXA10 and ITGB3 expression increased.At the same time,there was a difference in the expression of endometrial miRNA between implantation group and control group.Suggesting that metformin may improve the expression of HOXA10 and ITGB3 in the endometrium of PCOS patients by miRNA regulation of endometrial receptivity.MiR-1910-3p and miR-491-3p were predicted and screened for miRDB and TargetScan database,and related to HOXA10 and ITGB3.But what exactly is the specific mechanism by which miRNAs are involved in regulation is not yet clear.In this study,the effects of drugs on HOXA10 and ITGB3 expression were observed by using different concentrations of metformin and different time-acting endometrial epithelial cells Ishikawa.The expression of miR-1910-3p and miR-491-3p in the endometrium of PCOS and control group and its effect on HOXA10 and ITGB3 were studied.The results showed that the expression of miR-1910-3p and miR-491-3p was the basis for the evaluation of endometrial receptivity Clinical treatment to provide new ideas.Objective1.Effects of metformin on the expression of Ishikawa HOXA10 and ITGB3 in endometrial epithelial cells under different concentration and time of action;2.To detect the expression of miR-1910-3p and miR-491-3p in PCOS groupand control group of endometrial epithelial cells.3.To investigate the effects of miR-1910-3p and miR-491-3p on the expression of Ishikawa HOXA10 and ITGB3 in endometrial epithelial cells,and to explore the relationship between miR-1910-3p and miR-491-3p and HOXA10 and ITGB3.Methods1.The effect of metformin on Ishikawa HOXA10 and ITGB3 protein expression in endometrial epithelial cells was detected by Western-blotting at different concentrations and time of action.2.The endometrial epithelial cells were isolated from the endometrial tissue,and the miR-1910-3p and miR-491-3p were quantitatively analyzed by real time-PCR in the treatment group and control group in the expression.3.Overexpression of Ishikawa cell models were established by transfection of mimic of miR-1910-3p and miR-491-3p.The effects of changes in miR-1910-3p and miR-491-3p expression on endometrial markers HOXA10 and ITGB3 were detected by RT-PCR and Western-blotting.Results1.The expression of HOXA10 and ITGB3 in endometrial epithelial cells was increased in a dose-dependent manner(P<0.05).Compared with untreated cells,the expression of HOXA10 and ITGB3 was significantly increased at 12 h,24h and 48 h after treatment with metformin(P<0.05),but there was no obvious time-dependent relationship.2.Real time-PCR showed that the expression of miR-1910-3p and miR-491-3p in the endometrium of PCOS patients was significantly lower than that of the control group(P<0.05).3.The mRNA and protein expression levels of HOXA10 and ITGB3 were lower in the Ishikawa cells than those in the control group(P <0.001),while the expression of miR-1910-3p mimetic,HOXA10 mRNA and protein were transfected with miR-491-3p mimetic(P <0.001),but there was no significant change in ITGB3level(P> 0.05).ConclusionsMetformin significantly increased the expression of HOXA10 and ITGB3 in endometrial epithelial cells of Ishikawa;miR-1910-3p and miR-491-3p inhibited the expression of HOXA10 and ITGB3 in endometrial epithelial cells Ishikawa;metformin improved endometrial tolerance Sexual process may exist miR-1910-3p,miR-491-3 involved in regulation.