Hydrogen Sulfide Regulates The Decidualization Of Human Endometrial Stromal Cells And Its Biological Characteristics

Objective: Embryo implantation is a series of complex physiological processeswhich from the blastocyst implantation to placenta formation. In this process, embryonicdevelopment to the blastocyst begin stage implantation, deciduad endometriumproliferation and differentiation to the state of receptivity, mutual cooperation between theembryo and the mother. A variety of cytokines, transcription factors and hormones actingon the process, Embryo implantation failure is a key factor of the early abortion and thefailure transplant in invitrofertilization-embryo transfer(IVF-ET).Currently the molecular mechanisms studies in the process of embryo implantation not yet very clear. In terms of mother, on the one hand Decidualization endometrial stromal cells with a variety of cytokines, growth factors and hormones regulate embryonic extravillous trophoblast cells express a variety of genes, includingprostaglandin E2can promote uterine endometrial stromal cells decidualization in embryo implantation. On the other hand, the migration and invasion of endometrial stromal cells can enhance the migration and invasion of trophoblast cells. They can form a good interaction that ing blastocyst implantation. Therefore,the decidualization、migration and invassion of endometrial stromal cells plays a very important rolein embryo implantation process.Hydrogen sulfide (hydrogen sulfide, H2S) is a gaseous signaling molecule inthe body, which have some functions in a variety of physiological and pathologicalprocesses. Studies have shown that H2S as inflammatory cytokines involved in inflammation, regulating the function of vascular smooth muscle,and nervous system, and also plays an important role in the female reproductive system.In this study,we investigate the roles of H2S in the decidualization and the biological characteristics of human endometrial stromal cells.Thus to explore its effecton embryo implantation.Methods:1、the roles of H2S in the decidualization of human endometrial stromal cells(ESCs). To etect the expression of CBS and CSE which is the synthesis enzymes of H2S in the human endometrial tissue by using immunohistochemistry and Western-blot.The results showed that the expression of CBS is higer than CSE,and the endometrial tissue can produce H2S. We isolate, culture and identificateprimary human endometrial stromal cells, and build the decidualed human endometrial stromal cells model in vitro. We detect the expression of CBS and CSE in human endometrial stromal cells by Western-blot, founding that the expression of CBSincreased in the process of the decidualization, suggesting that H2S involved in the decidualization of human endometrial stromal cells. To study the role of H2S inthe decidualization of human endometrial stromal cells, we added AOAA which isthe inhibitor of CBS into the process of the decidualization of human endometrialstromal cells. Using Real-Time PCR to detect the the expression PRL and IGFBP1mRNA,which is the marker of decidualization, ELISA was also used to detect itsexpression in the cell supernatant.the results suggest that AOAA can reduce the decidualization of ESCs. Meanwhile, we used CBS siRNA transfect the endometrialstromal cells to interfere its expression of CBS which can reduce the synthesis ofH2S, Real-TimePCR and ELISA was used to detect the expression of PRL and IGFBP1, the results show that the decidualization reduced of ESCs reduced. That candemonstrate that H2S can promot the the decidualization of ESCs.2、The role of H2S in generation of PGE2which in the decidualization ofESCs. we added AOAA into the process of the decidualization of ESCs. To detectthe the expression of COX-2protein by Western-blot,which is the sythase of PGE2, ELISA was also used to detect the expression of PGE2in the cell supernatant.the results suggest that AOAA can reduce the the expression of COX-2and PGE2. Mean while, we added CBS siRNA into the process of the decidualization of ESCs. The expression of COX-2and PGE2also decreced.The results show that H2Scan promot the generation of PGE2in the decidualization of ESCs. Because PGE2can promote the decidualization of ESCs, so we believe H2S by upregulating thesynthesis of PGE2to promote the decidualization of ESCs by upregulating the generation of PGE2.3、The role of H2S in the biological characteristics of ESCs in decidualization. We added AOAA into the decidualization of ESCs，then examine the proliferation,migration and the invasion of ESCs by MTT,wound healing assay and transwellassay.The results show that the proliferation of the ESCs has no siginificant change,the motility and invasion decreased.We also added CBS siRNA into the process ofthe decidualization of ESCs. The results show that the motility and invasion also decreased. These results suggest that H2S can promote the migration and invasion of ESCs in the decidualization.Conclusions: This study demonstrated that H2S may promote the decidualizationof ESCs by upregulating the synthesis of prostaglandin E2, H2S can enhance themigration and invasion of decidual endometrial stromal cells. It is suggested that H2S play a important role in the process of embryo implantation. It can also provide a theoretical basis for the clinical research on early spontaneous abortion and failure transplant of IVF-ET.