TACE在着床窗口期小鼠子宫内膜上皮细胞中的表达及其抗体对胚泡粘附的影响  

Expression of tace in mouse uterine epithelial cell during implantation window period and effect of tace antibody on the embryo implantation

作　　者：刘帅[1] 姜蓉[2] 郗强[1] 杨戎[1] 张文文[1] 

机构地区：[1]重庆医科大学公共卫生学院生殖生理教研室,重庆400016 [2]重庆医科大学干细胞与组织工程研究室,重庆400016

出　　处：《重庆医科大学学报》2010年第8期1133-1137,共5页Journal of Chongqing Medical University

基　　金：重庆市科委自然科学基金项目(编号:CSTC;2007BB5273)

摘　　要：目的:通过检测肿瘤坏死因子α转换酶(Tumor necrosis factor α converting enzyme,TACE)在小鼠子宫内膜上皮细胞中的表达及其在体外着床模型中对胚泡粘附的影响,初步探讨TACE在小鼠生殖中的作用以及其与胚胎着床的关系。方法:体外培养孕4d(d4)小鼠子宫内膜上皮细胞和胚泡,构建胚泡体外着床模型;通过细胞免疫组化鉴定小鼠子宫内膜上皮细胞;运用RT-PCR和细胞免疫组化分别检测小鼠子宫内膜上皮细胞中TACE mRNA和蛋白的表达;向体外着床模型中添加不同浓度的TACE抗体,观察对胚泡体外着床的影响,并用细胞免疫化学检测抗体干预后各培养体系中TACE蛋白质的表达情况。结果:原代培养小鼠子宫内膜上皮细胞生长旺盛,鉴定结果显示:上皮细胞对角蛋白抗体有棕黄色阳性表达,对波形蛋白抗体为阴性表达。PCR结果显示TACE mRNA在共培养体系中有高表达,免疫组化结果与RT-PCR结果相一致;小鼠胚泡体外共培养具有较高的粘附率,添加TACE抗体后胚泡帖附率均显著低于对照组帖附率(P<0.05),且随着抗体浓度的降低胚泡粘附率随之升高,免疫化学检测结果也显示随着抗体浓度的降低TACE的表达量随之升高。结论:TACE可能参与了胚泡着床过程,有利于小鼠胚胎粘附、扩展,为胚胎的着床做准备。Objective：To detect expression of tumor necrosis factor-converting enzyme（TACE）on the mouse uterine epithelial cell and effect of implantation in vitro. To analyze its role in blastocyst implantation and to investigate the relation TACE withimplantation of mouse embryo. Methods：Co-culturing endometrial epithelial cells and blastocyst of the pregnant 4-day mouse to establish the implantation model in vitro. Epithelial cells was identified by Immunocytochemistry. RT-PCR and Immunocytochemistry were used to detect the expression of TACE gene and protein in the endometrial epithelial cells. Different concentrations of TACE antibody were added to the implantation model to observe effect of TACE antibody on mouse blastocyst implantation in vitro. Expression of TACE protein was detected by Immunocytochemistry in each antibody interference system. Results：Endometrial epithelial cells growed vigorously in vitro. Keratin antibody showed positive expression,meanwhile,Vimentin antibody showed negative expression in the endometrial epithelial cells. PCR showed that the expression of TACE gene was high and results of Immunocytochemistry was consisted with PCR. The adhesion rate of mouse blastocyst was high in the co-culture system and decreased remarkably when TACE antibody was added （P0.05）. The adhesion rate of mouse blastocyst appeared a increased trend as the concentration of TACE antibody lowed. Immunocytochemistry also showed that expression of TACE protein rised as the concentration of TACE antibody lowed. Conclusion：We presumed that TACE participated in the process of mouse blastocyst implantation,which promoted adhesion and outgrowth of blastocyst to prepare forblastocyst implantation.

关 键 词：TACE 小鼠体外着床 子宫内膜上皮细胞 原代培养 

分 类 号：R714.12[医药卫生—妇产科学]