摘要
目的观察过度训练大鼠肾组织Bax、Bcl-2及caspase-3的表达及其与肾小管上皮细胞凋亡的关系，探讨caspase依赖的凋亡信号途径在其中的作用及其机制。方法将48只雄性Wistar大鼠按随机数字表法分为对照组（CN）、力竭运动组（ES）、旋覆花素下预组（IB）。CN组为安静对照；ES组又根据力竭后恢复时间分为力竭后即刻（ESI）、力竭后6h（Es6h）和力竭后24h（ES24h）组；IB组于力竭运动前24h给予旋覆花素25ml／kg分3次灌胃后进行力竭运动，分为IB6h和lB24h组。采用大鼠游泳至力竭建直过度训练模型。TUNEL法检测肾小管上皮细胞凋亡。免疫组织化学法检测肾组织Bax、Bcl-2及easpase-3的表达。Western印迹法测定肾组织caspase-3蛋白的表达。用图像分析系统测定肾小管凋亡细胞、Bax、Bcl-2及caspase-3表达的平均吸光度并计算Bax和Bcl-2的比值。用Pearson法分析Bax和Bel-2的比值与easpase-3之间的相关性。用非参Speamlan法分析Bax／Bcl-2的比值和caspase-3与细胞凋亡之间的相关性。结果TUNEL法显示，过度训练大鼠肾组织凋亡细胞主要分布在肾小管上皮细胞，力竭后即刻、6h及24h肾小管上皮细胞凋亡数呈进行性增多（P〈0．01）；免疫绀化显示，过度训练大鼠肾组织caspase-3的表达及分布与Bax／Bcl-2比值变化及凋亡细胞的分布-敛。图像分析最示，大鼠肾小管七皮细胞Bax／Bcl-2比值与easpase-3的表达于力竭后即刻、力竭后6h、力竭后24h逐渐增高（均P〈0．05），均显著高于对照组（均P〈0．05）。Western印迹测定结果也屁示easpase-3蛋白表达的变化趋势。过度训练大鼠肾小管E皮细胞Bax／Bel-2比值与easpase-3的表达呈正相关（r=0．865，P〈0．05）；Bax／Bel-2比值和easpase-3的表达与肾小管上皮细胞凋亡之间均呈正相关（r=0．674，r=0．837，均P〈0．05）。用旋覆花素干预后，过度训练引起的肾小管上皮细胞Bax�
Objective To observe the expression changes of renal tissue Bax, Bcl-2 and caspase-3, to wxamine the correlation between the ratio of Bax to Bcl-2, caspase-3 and renal tubular cells apoptosis, and to investigate the role of caspase-related signal pathway. MethodsForty-eight male Wistar rats were randomly divided into three groups： control （CN, n=8）, exhaustive swimming （ES, n=24） and inula britannica （IB, n=16） group. The rats of CN were quiet without swimming. The rats of ES swam to exhaustive state and were sacrificed at immediately（ESI）, 6 hour （ES 6 h） and 24 hour （ES 24 h） after exhaustive swimming respectively. The rats of IB took orally inula britannica at the dose of 25 ml/kg body weight at 24 h before swimming and then swam to exhaustive state. The rats of IB group were sacrificed at 6 hour （IB 6 h） and 24 hour （IB 24 h） after exhaustive swimming. The animal model of overtraining-induced acute kidney injury was developed by exhaustive swimming. The renal cell apoptosis was measured by the method of TUNEL. The expressions of Bax, Bcl-2, caspase-3 in renal tissue were observed by immunohistochemistry. The expression of caspase-3 protein was examined by Western blotting. The correlation between the ratio of Bax to Bcl-2 and caspase-3 was analysed by Pearson method, and the correlation between the ratio of Bax to Bcl-2, caspase-3 and renal tubular cell apoptosis was analysed by Spearman method. Results The number of renal tubular apoptotic cells was increased progressively in ESI to ES 24 h rats by TUNEL （P〈0.05）. Immuuohistochemistry staining showed that the ratio of Bax to Bcl-2 and caspase-3 in renal tubular cells were increased progressively at 0 h, 6 h and 24 h after exhaustive swimming compared with control group （P〈 0.05）. The change of renal tissue caspase-3 was also revealved by Western blotting analysis. The ratio of Bax to Bcl-2 and caspase-3 in renal tubular cell was correlated positively （r=0.865, P〈 0.05）, The ratio of Bax to Bcl-2, and caspas
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2011年第2期118-123,共6页
Chinese Journal of Nephrology
基金
全军“十一五”医药卫生科研基金（06MA071）
