抗Thy1系膜增殖性肾炎大鼠模型中G0/G1期细胞周期蛋白表达变化

The changes of G0/G1 phase protein expression in the rat model of anti-Thy1 mesangial proliferative glomerulonephritis

目的:探讨抗Thy-1系膜增殖性肾炎大鼠模型中各病理时间点G0/G1期细胞周期蛋白变化。方法:利用OX-7杂交瘤细胞株产生抗Thy1抗体,纯化后经尾静脉注射Wistar大鼠建立抗Thy1系膜增殖性肾炎大鼠模型。用免疫组化方法,检测各病理时间点(第0、3、5、7、10、14d)Ki-67及细胞周期蛋白cyclin D1的表达变化趋势;Western Blot检测各实验组时间点G0/G1期细胞周期相关蛋白cyclin D1、cyclin E、CDK2、CDK4、CDK6、p27kip1表达变化趋势。结果:PAS染色显示各时间点Ki-67表达与病理变化正相关。与对照组比较,病理组d7Ki-67表达量显著增加(P<0.05),并随病理转归而下降。细胞周期蛋白cyclin D1的表达与Ki-67趋势相同,在病理最终的d7表达量最高(P<0.05),随病理转归而回到正常。Western Blot检测病理组细胞周期相关蛋白,与对照组相比,病理组d7,cyclin D1、cyclin E、CDK2、CDK4、CDK6的表达最高(P<0.05),p27kip1表达量最低(P<0.05)。结论:在抗Thy-1系膜增殖性肾炎大鼠模型中证实,G0/G1期细胞周期蛋白与病理变化正相关。

Objective：To explore the change of G0/G1 phase proteins in the different time points of pathological stages in the rat model of anti-Thy-1 mesangial proliferative glomerulonephritis.Methods：We established the anti-Thy1 mesangial proliferative nephritis rats model by using the Wistar rats through injected the anti-Thy1 monoclonal antibody via the tail vein.The expression levels of Ki67 and cyclin D1 at different pathological time point（day 0,3,5,7,10,14）were detected by immune-histochemical methods.And the expression levels of G0/G1 phase of the cell cycle-related proteins,cyclin D1,cyclin E,CDK2,CDK4,CDK6,p27kip1 at different pathological time points were measured by the Western Blot method.Results：There was a positive relationship between the expression of Ki67 and the pathological changes.Compared with the control group,the expression of Ki67 in the model group increased significantly at the 7th day（P〈0.05）,then decreased with pathological development.There was a same trend in the expression of cyclin D1 as Ki67.The expression of cyclin D1,cyclin E,CDK2,CDK4,CDK6 reached to the highest level at the 7th day compared with the control group（P〈0.05）,while the expression of p27^kip1 reduced to the lowest level（P〈0.05）.Conclusion：This study confirmed that the pathological changes was positively associated with the changes of G0/G1 phase protein of the cell cycle in the anti-Thy1 mesangial proliferative glomerulonephritisrat model.