过度训练大鼠肾组织细胞凋亡的基因调控机制及山莨菪碱干预的影响

Gene regulation of renal cell apoptosis and effect of anisodamine on overtrained rat by exhausting swimming

目的研究过度训练大鼠肾组织细胞Bax和Bcl-2的表达以及Bax/Bcl-2比值与肾组织细胞凋亡的关系,观察山莨菪碱对上述蛋白表达的影响,探讨肾组织细胞凋亡的基因调控及山莨菪碱抗肾组织细胞凋亡的分子机制。方法采用大鼠游泳至力竭建立过度训练模型,将大鼠随机分为3组:安静对照组(CN,n=8)、力竭运动组(分为ESI、ES 6h、ES 24h 3个时间点,每个时间点8只动物)、山莨菪碱防治组(分为AD6h、AD24h 2个时间点,每个时间点8只动物)。采用免疫组织化学方法检测大鼠肾组织Bax、Bcl-2蛋白的表达;采用CMIAS病理图像分析仪测量Bax、Bcl-2蛋白表达的平均光密度;对各组大鼠肾组织Bax/Bcl-2比值与肾组织细胞凋亡率进行相关性分析。结果与CN组相比,ESI、ES 6h、ES 24h组大鼠肾组织Bax的表达逐渐增强,Bcl-2的表达逐渐减弱,Bax/Bcl-2比值呈进行性增高(P<0.05)。AD6h、AD24h组大鼠与同期力竭组比较,Bax的表达减弱,Bcl-2的表达增强,Bax/Bcl-2比值降低(P<0.05)。各组大鼠肾组织Bax/Bcl-2比值与肾组织细胞凋亡率呈明显的正相关(r=0.674,P<0.01)。结论Bax、Bcl-2及Bax/Bcl-2比值作为调控细胞凋亡的中心环节共同参与过度训练致肾组织细胞凋亡发生的调控,山莨菪碱可通过调节Bax、Bcl-2及Bax/Bcl-2比值起到抗肾组织细胞凋亡的作用。

Objective To study the expressions of apoptosis related genes Bcl-2 and Bax proteins in rafs renal tissue and the rela tionship between the ratio of Bcl-2/Bax and renal cell apoptosis induced by overtraining, and observe the effect of anisodamine on the expression of genes Bcl-2 and Bax in exhausted rats. Methods The animal model of acute kidney injury induced by exhausting swimming was reproduced. Forty eight male Wistar rats were randomly assigned imo sedentary control group （CN, n=8）, exhausting swimming group （immediate, 6 hours and 24 hours after exhausting swimming, ESI, ES6h and ES24h, 8 each）, Anisodamine group （6 hours and 24 hours after exhausting swimming, AD6h and AD24h, 8 each）. The expressions of Bcl-2 and Bax were detected by immunohistochemical staining and image analyzer. The correlation between the ratio of Bcl-2/Bax and renal cell apoptosis was analyzed. Results The analysis showed that the expression of Bax increased, and of Bcl-2 decreased, the ratio of Bax/Bcl-2 increased remarkably in ESI group compared with CN group （P〈0.05）. It was more prominent in ES6h group and reached the top level in ES24h group. The expression of Bax decreased remarkably, and of Bcb2 increased significantly, the ratio of Bax/Bcl-2 decreased remarkably in AD6h and AD24h groups compared with that of F, S6h and ES24h groups. There was a significant positive correlation between the value of Bax/Bcl 2 and the ratio of renal cell apoptosis （r=0. 674, P〈0.01 ）. Conclusion Renal cell apoptosis in exhausting rats may be determined by down-regulation of Bcl-2 and up-regula tion of Bax expressions and the value of Bax/Bcl-2. Anisodamine can inhibit call apoptosis by regulating the expression of Bax, Bcl-2 and the value of Bax/Bcl-2.