TACE在著床窗口期小鼠子宮內(nèi)膜上皮細(xì)胞中的表達(dá)及其抗體對胚泡粘附的影響
發(fā)布時間：2019-03-08 22:02
【摘要】：目的:通過檢測腫瘤壞死因子α轉(zhuǎn)換酶(Tumor necrosis factor α converting enzyme,TACE)在小鼠子宮內(nèi)膜上皮細(xì)胞中的表達(dá)及其在體外著床模型中對胚泡粘附的影響,初步探討TACE在小鼠生殖中的作用以及其與胚胎著床的關(guān)系。方法:體外培養(yǎng)孕4d(d4)小鼠子宮內(nèi)膜上皮細(xì)胞和胚泡,構(gòu)建胚泡體外著床模型;通過細(xì)胞免疫組化鑒定小鼠子宮內(nèi)膜上皮細(xì)胞;運用RT-PCR和細(xì)胞免疫組化分別檢測小鼠子宮內(nèi)膜上皮細(xì)胞中TACE mRNA和蛋白的表達(dá);向體外著床模型中添加不同濃度的TACE抗體,觀察對胚泡體外著床的影響,并用細(xì)胞免疫化學(xué)檢測抗體干預(yù)后各培養(yǎng)體系中TACE蛋白質(zhì)的表達(dá)情況。結(jié)果:原代培養(yǎng)小鼠子宮內(nèi)膜上皮細(xì)胞生長旺盛,鑒定結(jié)果顯示:上皮細(xì)胞對角蛋白抗體有棕黃色陽性表達(dá),對波形蛋白抗體為陰性表達(dá)。PCR結(jié)果顯示TACE mRNA在共培養(yǎng)體系中有高表達(dá),免疫組化結(jié)果與RT-PCR結(jié)果相一致;小鼠胚泡體外共培養(yǎng)具有較高的粘附率,添加TACE抗體后胚泡帖附率均顯著低于對照組帖附率(P0.05),且隨著抗體濃度的降低胚泡粘附率隨之升高,免疫化學(xué)檢測結(jié)果也顯示隨著抗體濃度的降低TACE的表達(dá)量隨之升高。結(jié)論:TACE可能參與了胚泡著床過程,有利于小鼠胚胎粘附、擴展,為胚胎的著床做準(zhǔn)備。
[Abstract]:Objective: to detect the expression of tumor necrosis factor 偽 converting enzyme (Tumor necrosis factor 偽 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.
【作者單位】： 重慶醫(yī)科大學(xué)公共衛(wèi)生學(xué)院生殖生理教研室;重慶醫(yī)科大學(xué)干細(xì)胞與組織工程研究室;
【基金】：重慶市科委自然科學(xué)基金項目(編號:CSTC,2007BB5273)
【分類號】：R714.12
本文編號：2437244
[Abstract]:Objective: to detect the expression of tumor necrosis factor 偽 converting enzyme (Tumor necrosis factor 偽 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.
【作者單位】： 重慶醫(yī)科大學(xué)公共衛(wèi)生學(xué)院生殖生理教研室;重慶醫(yī)科大學(xué)干細(xì)胞與組織工程研究室;
【基金】：重慶市科委自然科學(xué)基金項目(編號:CSTC,2007BB5273)
【分類號】：R714.12
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1 楊潔玉;TACE在動情周期和早孕小鼠子宮內(nèi)膜的表達(dá)及作用研究[D];重慶醫(yī)科大學(xué);2008年
，本文編號：2437244
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