孕酮对奶牛子宫内膜上皮细胞Wnt/β-catenin信号通路关键蛋白表达的影响

Effects of Progesterone on the Expression of Wnt/β-catenin Signaling Pathway in Endometrial Epithelial Cells of Dairy Cows

为研究孕酮对奶牛子宫内膜上皮细胞Wnt/β-catenin信号通路关键蛋白表达的影响,本试验以原代奶牛子宫内膜上皮细胞为材料,添加不同浓度孕酮(0、1、3、5 ng/mL)对其培养,采用CCK-8法检测孕酮对细胞增殖的影响;免疫印迹法检测孕酮对Wnt/β-catenin信号通路关键蛋白表达的影响。结果表明:与对照组相比,添加1、3 ng/mL孕酮组奶牛子宫内膜上皮细胞增殖率提高(P>0.05),5 ng/mL孕酮组细胞增殖率降低(P>0.05);与对照组相比,1 ng/mL和3 ng/mL孕酮组Wnt/β-catenin信号通路关键蛋白β-catenin、cyclin D1和c-Myc的表达水平均升高(P<0.01),而5 ng/mL孕酮则抑制了β-catenin的表达(P<0.05),且cyclin D1和c-Myc蛋白表达较3 ng/mL孕酮组有下降趋势(P>0.05)。综上,高浓度孕酮抑制了体外培养的原代奶牛子宫内膜上皮细胞Wnt/β-catenin信号通路的激活和表达,提示该信号通路参与了奶牛产后子宫复旧延迟进程,为进一步研究奶牛子宫内膜的发病机制提供思路。

Endometritis in dairy cows is one of the important clinical diseases.In this reseach,the primary endometrial epithelial cells of diary cows were used as materials to simulate the progesterone level in peripheral blood of postpartum dairy cows.The effects of progesterone in different concentrations(0、1、3、5 ng/mL)on proteins expression in Wnt/β-catenin signaling pathway were studied.The proliferation and the expression of key proteins of endometrial epithelial cells induced by progesterone were detected with CCK-8 and western blot methods respectively.Results showed that 1 and 3 ng/mL progesterone groups promoted the proliferation rate of bovine endometrial epithelial cells.5 ng/mL progesterone decreased the proliferation rate of the cells,but the difference was not significant(P>0.05),compared with the control group(0 ng/mL).1 ng/mL and 3ng/mL progesterone significantly promoted the expression of key proteins in Wnt/β-catenin signaling pathway,and the expression ofβ-catenin,cyclin D1 and c-Myc protein increased significantly,compared with the control group(P<0.01).While 5 ng/mL progesterone significantly inhibitedβ-catenin expression(P<0.05),and the expression of cyclin D1 and c-Myc proteins decreased,compared with the 3 ng/mL progesterone group,which was not significant(P>0.05).In summary,progesterone in high concentrations inhibited the activation and expression of Wnt/β-catenin signaling pathway in primary endometrial epithelial cells of dairy cows,which verified that Wnt/β-catenin signaling pathway was involved in the delayed progression of postpartum uterine involution in dairy cows.The research provides new idea for further study on the pathogenesis of bovine endometrium.