| ??ObjectiveEmbryo implantation is the key to the success of pregnancy.During embryo implantation,endometrium is decidualized under the action of many pregnancy-related factors,which is crucial for embryo implantation,pregnancy establishment and maintenance.Decidualization is characterized by increased endometrial glands,recasting of spiral arteries,recruitment of immune cells,and changes in extracellular matrix components.Another extremely important characteristic is the transformation of endometrial stromal cells into decidual cells,marked by increased secretion of decidual marker prolactin(PRL)and insulin-like growth factor-1(IGFBP-1).Protein glycosylation is an important post-translational processing and modification of proteins,which plays an important role in a variety of physiological and pathological processes,including embryo implantation,cell adhesion,malignant transformation,inflammation,signal transduction.An abnormal glycosylated protein molecule with altered biological properties and functions.Fucosylation is an important form of protein glycosylation modification.in this paper,fucosyltransferase?(FUT7)belong to the N-fucosetransferase,FUT7 through?1,3-glycosidic bond specific catalytic synthesis of sialic acid Louis oligosaccharides antigen-X(sialyl Lewis X,sLe~X).Previous studies have found that FUT7 plays an important role in embryo implantation,such as promoting the adhesion between the embryo and endometrial epithelial cells.However,the role of endometrium decuvialization and its mechanism are still unknown.Therefore,this paper aims to explore the role of FUT7 in the process of endometrial decidua,so as to provide a theoretical basis for understanding the causes of female abortion.??Methods1.Immunohistochemistry,Western blot and Real-time PCR were used to detect the expression of FUT7 and sLe~Xin human endometrium during proliferative phase,secretory phase,early pregnancy and endometrium of patients with inevitable abortion.2.Induced decidualization of endometrial stromal cells(HESCs)in vitro under the combined action of Medroxyprogesterone 17-acetate(MPA)and dibutyryl c AMP(dbc AMP).The expression of FUT7 before and after decidual inducing was detected by Western blot,Real-time PCR and immunofluorescence.3.The expression of FUT7 was up-regulated/down-regulated,and the expression of decidual markers PRL,IGFBP-1 and sLe~Xwas detected by Western blot,Real-time PCR and immunofluorescence.4.The expression of FUT7 was up-regulated/down-regulated,and the proliferation ability of decidual cells was detected by CCK8,plate cloning,Western blot and Real-time PCR.5.Trophoblast cells(HTR)induced endometrial stromal cells to simulate embryo implantation.Western blot and Real-time PCR were used to detect that FUT7 promoted HESCs decidualization by activating the ERK1/2 signaling pathway.6.In animal experiments,Immunohistochemistry experiment detected the expression of PRL,FUT7 and sLe~Xin the endometrium of mouse with different days of pregnancy.FUT7-si RNA was injected into the uterine horn and normal saline on the other side as control,The expression of FUT7,sLe~X,PRL,IGFBP-1 and PCNA as well as ERK1/2and p-ERK1/2 signaling pathway in the endometrium of mice in different treatment groups were detected by Immunohistochemistry experiment.??Results1.Immunohistochemical results showed that the expression of FUT7 and sLe~Xincreased in the proliferative phase,secretory phase and early pregnancy of human endometrial tissue,and significantly decreased in the inevitable abortion endometrial tissue.2.HESCs were induced to decidualization in vitro.Western blot,Real-time PCR and immunofluorescence confirmed that the decidual markers PRL and IGFBP-1 increased significantly after decidualization,indicating that the decidualization was successfully induced in vitro.The expression of FUT7 was increased significantly after decidualization.3.The expression of FUT7 had been up-regulated or down-regulated,and the cells were induced to decidualization at the same time.The results of Western blot,Real-time PCR and immunofluorescence showed that the up-regulated expression of FUT7 could promote the expression of the decidual marker PRL and IGFBP-1.Down-regulated expression of FUT7 could inhibit its expression.4.The results of plate cloning and CCK8 experiment showed that after decidualization,up-regulating the expression of FUT7,the proliferation ability was significantly increased,while down-regulating the expression of FUT7,the proliferation ability was decreased.Western blot and Real-time PCR experiments showed that up-regulation of FUT7 expression increased the expression levels of cell proliferation nuclear antigen(PCNA)and cyclin D,cyclin E,CDK4 and CDK6,and decreased the expression levels of antiproliferation markers(p21 and p27).When the expression of FUT7 was down-regulated,the above indicators also changed accordingly.This indicated that FUT7 promoted cell proliferation by activating cyclin proteins.5.The results of Western blot showed that p-ERK1/2 signal path was activated under the stimulation of HTR cells.After artificial induction of HESCs decidualization,and up-regulated the expression of FUT7,p-ERK1/2 was activated,down-regulated the expression of FUT7,the p-ERK1/2 was inhibited.6.Immunohistochemical results showed that the expressions of PRL,FUT7 and sLeX increased in the endometrium of mice on the days(4-6)of pregnancy.After FUT7-si RNA injection,endometrium decidualization was inhibited and embryo implantation rate was reduced.Immunohistochemical analysis showed that down-regulation of FUT7 could inhibit the occurrence of decidualization,the proliferation of cells and the activation of p-ERK1/2 signaling pathway.??Conclusion1.The expressions of FUT7 and sLe~Xin the decidual tissues of early pregnancy women were increased,while decreased in the inevitable abortion patients.2.FUT7 regulates the synthesis of sLe~Xand promotes the proliferation of stromal cells and affects decidualization of stromal cells.3.Down-regulation of FUT7 hinders the process of uterine decidualization and inhibit embryo implantation in mice. |
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