TGF-β1通过p38MAPK信号通路调节大鼠睾丸支持细胞闭锁蛋白表达的研究

TGF-β1 regulates the expression of occludin protein in rat sertoli cell by p38MAPK signaling pathway

目的探讨TGF-β1介导的p38MAPK信号通路对大鼠睾丸支持细胞闭锁蛋白表达的作用及意义。方法体外分离培养大鼠睾丸支持细胞,随机分为空白对照组、刺激组、受体阻滞剂组、受体阻滞剂+刺激组、抑制因子组和抑制因子+刺激组6组;测定各组细胞增殖情况、p38MAPK mRNA和闭锁mRNA的表达水平以及各组细胞闭锁蛋白和P-p38MAPK蛋白表达水平。结果组间比较差异有统计学意义(F=4.86,P<0.05)。刺激组细胞增殖活性升高,差异有统计学意义(P<0.05);刺激组闭锁蛋白mRNA表达比空白对照组降低,p38MAPK mRNA表达升高;抑制因子组p38MAPK mRNA表达降低,差异均有统计学意义(P<0.05)。受体阻滞剂+刺激组闭锁蛋白mRNA表达比刺激组升高,p38MAPK mRNA表达降低;抑制因子+刺激组闭锁蛋白mRNA表达升高,p38MAPK mRNA表达降低,差异均有统计学意义(P<0.05)。刺激组闭锁蛋白表达比对照组降低,P-p38MAPK蛋白表达升高;抑制因子组P-p38MAPK蛋白表达降低,差异有统计学意义(P<0.05)。受体阻滞剂+刺激组闭锁蛋白表达比刺激组升高,P-p38MAPK蛋白表达降低;抑制因子+刺激组闭锁蛋白表达升高,P-p38MAPK蛋白表达降低差异均有统计学意义(P<0.05)。结论 TGF-β1可能通过p38MAPK信号通路调控大鼠睾丸支持细胞闭锁蛋白的表达。

Objective To investigate the role of TGF-β1 mediated p38MAPK signaling pathway in the expression of occludin protein in rat sertoli cell. Methods The rats sertoli cells were isolated and cultured in vitro,and then randomly assigned to blank control group,stimulus group,receptor blockers group,receptor blockers ＋ stimulus group,inhibitory factor group and inhibitory factor ＋ stimulus group. MTT method was used to determine the proliferation of rat sertoli cells in each group.RT-PCR was used to determine the expression of p38MAPK mRNA and occludin mRNA in each group. Occludin protein and p-p38MAPK protein were detected by Western-blot. Results The results of MTT showed that the difference between groups was statistically significant（ F = 4. 86,P〈0. 05）. The cell proliferation activity of stimulus group significantly increased compared with the other five groups（ P〈0. 05）. In stimulus group,the expression of occludin mRNA significantly decreased,and p38MAPK mRNA significantly increased,compared with the blank control group. In inhibitory factor group,p38MAPK mRNA significantly decreased compared with the blank control group（ P〈0. 05）. In the stimulus group,occludin mRNA significantly increased,with the difference statistically significant（ P〈0. 05）. mRNA increased but p38MAPK mRNA decreased in receptor blockers ＋ stimulus group,with the differences statistically significant（ P〈0. 05）. Compared with the control group,p38MAPK mRN increased in the stimulus group; in inhibitory group,p38MAPK mRN decreased,with the differences statistically significant（ P〈0. 05）. Compared with the stimulus group,P-p38MAPK protein significantly decreased in receptor blockers ＋ stimulus group. Compared with the stimulus group,P-p38MAPK protein significantly decreased in inhibitory factor ＋ stimulus group,with the differences statistically significant（ P〈0. 05）. Conclusion TGF-β1 may regulate the expression of occludin protein in rat sertoli cell by p38MAPK signaling pathway.