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
发布时间：2019-03-08 22:02
【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(P0.05),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。
[Abstract]:Objective: to detect the expression of tumor necrosis factor 伪 converting enzyme (Tumor necrosis factor 伪 converting enzyme,TACE in mouse endometrial epithelial cells and its effect on blastocyst adhesion in vitro implantation model. To explore the role of TACE in mouse reproduction and its relationship with embryo implantation. Methods: endometrial epithelial cells and blastocysts of mouse endometrial epithelial cells and blastocysts were cultured in vitro on the 4th day of gestation (d4) to construct blastocyst implantation model, and the mouse endometrial epithelial cells were identified by immunocytochemistry. The expression of TACE mRNA and protein in mouse endometrial epithelial cells were detected by RT-PCR and immunocytochemistry, respectively. The effects of different concentrations of TACE antibody on blastocyst implantation were observed in vitro implantation model, and the expression of TACE protein in different culture systems was detected by immunocytochemistry. Results: the primary cultured mouse endometrial epithelial cells grew exuberantly. The results showed that the positive expression of keratin antibody in epithelial cells was brown-yellow. The expression of vimentin antibody was negative. The results showed that TACE mRNA was highly expressed in co-culture system, and the results of immunohistochemistry were consistent with the results of RT-PCR. Mouse blastocyst co-culture in vitro had a higher adhesion rate, and the blastocyst adhesion rate after the addition of TACE antibody was significantly lower than that of the control group (P0.05), and the blastocyst adhesion rate increased with the decrease of antibody concentration. The results of immunohistochemistry also showed that the expression of TACE increased with the decrease of antibody concentration. Conclusion: TACE may be involved in the process of blastocyst implantation, which is beneficial to mouse embryo adhesion, expansion and preparation for embryo implantation.
【作者单位】： 重庆医科大学公共卫生学院生殖生理教研室;重庆医科大学干细胞与组织工程研究室;
【基金】：重庆市科委自然科学基金项目(编号:CSTC,2007BB5273)
【分类号】：R714.12
本文编号：2437244
[Abstract]:Objective: to detect the expression of tumor necrosis factor 伪 converting enzyme (Tumor necrosis factor 伪 converting enzyme,TACE in mouse endometrial epithelial cells and its effect on blastocyst adhesion in vitro implantation model. To explore the role of TACE in mouse reproduction and its relationship with embryo implantation. Methods: endometrial epithelial cells and blastocysts of mouse endometrial epithelial cells and blastocysts were cultured in vitro on the 4th day of gestation (d4) to construct blastocyst implantation model, and the mouse endometrial epithelial cells were identified by immunocytochemistry. The expression of TACE mRNA and protein in mouse endometrial epithelial cells were detected by RT-PCR and immunocytochemistry, respectively. The effects of different concentrations of TACE antibody on blastocyst implantation were observed in vitro implantation model, and the expression of TACE protein in different culture systems was detected by immunocytochemistry. Results: the primary cultured mouse endometrial epithelial cells grew exuberantly. The results showed that the positive expression of keratin antibody in epithelial cells was brown-yellow. The expression of vimentin antibody was negative. The results showed that TACE mRNA was highly expressed in co-culture system, and the results of immunohistochemistry were consistent with the results of RT-PCR. Mouse blastocyst co-culture in vitro had a higher adhesion rate, and the blastocyst adhesion rate after the addition of TACE antibody was significantly lower than that of the control group (P0.05), and the blastocyst adhesion rate increased with the decrease of antibody concentration. The results of immunohistochemistry also showed that the expression of TACE increased with the decrease of antibody concentration. Conclusion: TACE may be involved in the process of blastocyst implantation, which is beneficial to mouse embryo adhesion, expansion and preparation for embryo implantation.
【作者单位】： 重庆医科大学公共卫生学院生殖生理教研室;重庆医科大学干细胞与组织工程研究室;
【基金】：重庆市科委自然科学基金项目(编号:CSTC,2007BB5273)
【分类号】：R714.12
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1 杨洁玉;TACE在动情周期和早孕小鼠子宫内膜的表达及作用研究[D];重庆医科大学;2008年
，本文编号：2437244
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