The Effects Of Polycystic Ovary Syndrome On Immune Cells And Uterine Extracellular Matrix During Peri-Implantation

In 2018,"International Evidence-based Medicine Guide for Polycystic Ovary Syndrome" reported that 8%-13% of women suffered from polycystic ovary syndrome(PCOS),and more than 80% of PCOS patients in childbearing age suffered from infertility.Normal pregnancy requires the interaction between embryos with implantation ability and receptive endometrium,both of which are indispensable.Even if PCOS patients have transplanted high-quality embryos through assisted reproductive technology,the abnormality of endometrium will still lead to implantation failure or poor pregnancy outcome.At present,the research on PCOS endometrial abnormality is relatively insufficient,and the exact mechanism of implantation failure is still unclear.The study on the mechanism of endometrial changes in peri-implantation PCOS can help us find potential therapeutic targets,further improve the success rate of embryo implantation and improve pregnancy loss.This paper consists of three levels: mouse PCOS model,short-term intervention of dehydroepiandrosterone and human PCOS intima specimen,the changes of endometrial receptivity of PCOS during perinatal period were analyzed and explored.In this paper,the mouse PCOS model was established by dehydroepiandrosterone.These mice successfully showed PCOS characteristics such as hyperandrogenism,weight gain and infertility.Transcriptome sequencing was performed in the uterus of PCOS mice on the 4th day of gestation,610 genes were up-regulated and 91 genes were down-regulated.Bioinformatics analysis shows that: In terms of cellular components and molecular functions,the extracellular matrix(ECM)and a variety of related functions were the most abundant of differential genes,such as cell adhesion molecules basal membrane cell desmosome cell apical plasma membrane microvilli micro villi plasma membrane.The enrichment analysis of biological processes showed that the biological processes related to immune response accounted for the highest proportion,Including bacterial immune response and defense response and so on.To further clarify the effects of DHEA on mouse uterine extracellular matrix and immune cells,mice were treated with DHEA on day 3 of gestation,DHEA was administered daily on days 1 to3 of gestation and the pseudo-pregnant mice were treated with DHEA on the 1st-3rd day of pregnancy.In addition,ovariectomized mice were treated with DHEA for 21 days.Based on the above predictive analysis and different DHEA treatments,we studied in detail the number and location of immune cells in mouse uterus.(1)Compared with the control group,the number of neutrophils in the uterus of PCOS mice increased and gathered near the luminal epithelium during peri-implantation,the difference was extremely significant.The number of neutrophils in the uterus increased significantly after DHEA treatment on the 1st to 3rd day of gestation,and they clustered near the uterine cavity.Pseudo-pregnant mice were also treated with DHEA for 3 days,and the number of neutrophils increased,but the location of neutrophils did not change.(2)The number of macrophages in the uterus of peri-implantation PCOS mice was significantly increased and the macrophages gathered near the uterine cavity.DHEA treatment for 3days had little effect on the number and distribution of macrophages.However,longterm exposure to DHEA increases the number of macrophages in the intima and causes them to accumulate near the uterine cavity,and their distribution changes are not related to the embryo in uterine cavity.(3)The number of dendritic cells in the endometrial stroma of PCOS during peri-implantation was decreased,the number of dendritic cells in the muscular layer was not significantly changed.Dendritic cells in the uterus did not change significantly after short-term exposure to DHEA.The number of dendritic cells in uterine stroma of pseudo-pregnant mice increased significantly.The number of dendritic cells in uterine stroma and muscular layer of ovariectomized mice increased significantly after 21 days of DHEA treatment.(4)Compared with the control group,the number of mast cells increased in the myometrium of PCOS mice during periimplantation.DHEA treatment had little effect on the number of mast cells in uteri.(5)The number of B cells in the uterus of PCOS mice increased significantly during periimplantation,and the B cells gathered near the uterine epithelium.Treatment with DHEA for 3 days resulted in the increase of B cells and the accumulation of B cells into the uterine epithelium,but the intrauterine embryos had little effect on the distribution of B cells.In addition,we analyzed in detail the effects of DHEA on peri-implantation uterine intercellular junctions and extracellular matrix.(1)Compared with the control group,the expression of Ezrin in the uterine cavity and glandular epithelium of PCOS mice during peri-implantation period was significantly increased.Treatment with DHEA for either 1-or 3-days during peri-implantation significantly increased Ezrin expression in mouse uterine epithelium,and influence their patterns of expression.The effect of embryo on Ezrin expression in utero was not obvious.In the window period of human embryo implantation,the expression of Ezrin in endometrium and glandular epithelium of PCOS patients was higher than that of control group,and the difference was significant.(2)The expression of E-E-E-cadherin in the uterine cavity and glandular epithelium of PCOS mice was significantly increased during peri-implantation.DHEA treatment for 3 days had little effect on E-cadherin in utero.However,after short-term treatment with DHEA,the expression of E-cadherin in the uterine cavity and glandular epithelium of pseudo-pregnant mice increased.In the window period of human embryo implantation,the expression of E-cadherin in endometrium of PCOS patients was significantly higher than that of the control group,there was no significant difference in glandular epithelial E-cadherin expression between the two groups.(3)Compared with the control group,the expression of laminin in myometrium of PCOS increased significantly during peri-implantation.The deposition of laminin in the basal membrane of the luminal epithelium and in the basal membrane of the glandular epithelium and the vascular basal membrane was thin,from the normal serrated into a thin line.Treatment with DHEA for 3 days during peri-implantation changed the morphology of laminin deposition in the epithelial and vascular basement membranes.However,it has little effect on its thickness.At the same time,the expression of laminin in myometrium was significantly increased,and the embryo had no significant effect on the change of laminin in uterine myometrium during peri-implantation.(4)The expression of laminin? 1 in uterine stroma of PCOS mice decreased with the decrease of the number of stromal vessels during peri-implantation.DHEA treatment for 3 days significantly decreased the expression of laminin ? 1 in glandular epithelial basement membrane.During the window of human embryo implantation,there was no significant difference in the expression of laminin ? 1 between endometrial luminal epithelium and glandular epithelium in patients with PCOS.(5)Compared with the control group,the deposition of collagen IV in the luminal epithelial basement membrane or glandular epithelial basement membrane in the uterus of PCOS mice became thinner during periimplantation.With the decrease of blood vessels in uterine stroma,the expression of collagen IV decreased significantly.After 3 days of DHEA treatment,the change was similar to that of PCOS.(6)CD31 is mainly expressed in the cytoplasm of vascular endothelial cells,and the expression of CD31 is denser in the uterus of mice on the 4th day of pregnancy.However,the expression decreased in the uterus of PCOS mice,and the difference was extremely significant.CD31 in the uterus of a PCOS mouse shows a circular cut of blood vessels,the density decreased obviously,and more vessels were long and narrow in the section lumen,there are fewer of them.Uterine blood vessels were significantly reduced after 3 days of DHEA treatment,and the embryo may have an effect on the number of blood vessels in the peri-implantation period.(7)The expression of Ulex Europaeus Agglutinin(UEA)receptor in uterine cavity epithelium of PCOS mice during peri-implantation was stronger than that in control group.UEA receptors were not uniformly distributed in the two groups.There was no significant difference in the expression of UEA receptor in glandular epithelium.The UEA receptor was strongly expressed in the uterine epithelium treated with DHEA during periimplantation.After 3 days of DHEA treatment,the expression of UEA receptor in the luminal epithelium was decreased and showed inhomogeneity.UEA receptors were not expressed in the endometrial epithelium and glandular epithelium after ovariectomy.Embryos in the uterine cavity during peri-implantation may induce increased expression of UEA receptorsIn conclusion,there were significant differences in m RNA levels between PCOS mice and control mice on day 4 of pregnancy.The main changes were in immune response and extracellular matrix.DHEA has a significant effect on the number and distribution of immune cells,such as neutrophils,macrophages,dendritic cells,B cells and mast cells,in the peri-implantation uterus.This may have adverse effects on its normal immune function.Extracellular matrix is an important component of cell communication and cytoskeleton.DHEA significantly altered the expression levels of Ezrin and E-cadherin,which may affect the normal adhesion and polarity of cells.The effect of DHEA on laminin and its subtype collagen IV and CD31 may lead to the remodeling of cytoskeleton and the abnormal intercellular communication during embryo implantation.