The Investigation On The Correlation Between Endometriosis With Sromal Cell-derived Factor-1 And Its Receptor

Objective To explore the method of culture for human stromal cell of normal endometrium, eutopic endometrium and ectopic endometrium.Methods Enzymolyses,filtration ,sedimentation and pasted wall purification were used to isolate and culture the stromal cells of twelve samples of normal endometrium, nine sampls of eutopic endometrium and six samples of ectopic endometrium. These stromal cells were observed by light microscopy and identified by immunocytochemical staining uising mouse antihuman cytokeratin and vimentin antibody.Results Ten normal endometrial sampls, six eutopic endometrial sampls and four ectopic endometrial sampls are isolated and cultured successfully. The purity of stromal cells is 93% verified by cytokeratin and vimentin antibody staining, all of these tromel cells could be transfered for six to eight generations.Conclusion Highly purified stromal cells can be obtained through enzymolysis filtration, sedimentation and pasted wall ,purification, the cellular model will be helpful to investigate the pathogenesis of endometriosis and therapy of endometriosis. Objective:To investigate the effect of 17β-estradiol on expression of SDF-1/CXCR4 mRNA and protein by endometrial stromal cells of endometriosis and the role of SDF-1/CXCR4 in pathogenesis of endometriosis.Methods:Separate and culture the primary eutopic ESC of endometriosis in vitro, The cells were treated by 17β-estradiol (17β-E2 )with various concentrations(0,10-12,10-10,10-8,10-6 mol/L)for different time(0, 12, 24, 48h). The expressions of SDF-1/CXCR4 mRNA and protein in ESC of endometriosis were analyzed by Real-time quantitative PCR(RQ-PCR) and Flow Cytometry. Meanwhile, the expression of SDF-1/CXCR4 mRNA in cells by the ER inhibitor ICI182780(10-8 mol/L) and 17β-E2 treated were detected in the same waysResults:After treating of 17β-estradiol in different concentration for 12h,24h and 48h, the expression of SDF-1/CXCR4 mRNA and protein are both up-regulated to different extents , the role of 17β-estradiol is in evidence in concentration of 10-10mol/L. Theses kinds of roles of 17β-estradiol on SDF-1 and CXCR4ER can be blocked by ER inhibitor ICI182780 (10-8 mol/L) obviously.Conclusion:The expression of SDF-1/CXCR4 mRNA and protein can be promoted by 17β-estradiol in eutopic ESC of endometriosis. SDF-1 and its receptor CXCR4 may have correlation with the development of endometriosis.