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大鼠肾脏缺血再灌注损伤后自噬的初步研究
Preliminary study on autophagy after renal Ischemia reperfusion injury in rats

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DOI:
作者:
蔡彦，叶朝阳，高翔，付莉莉，刘晔，李楠，王文苓，梅长林
CAI Yan，YE Chaoyang，GAO Xiang，FU Lili，LIU Ye，LI Nan，WANG Wenling，MEI Changlin
作者单位:
上海长征医院肾内科
Department of Nephrology, Shanghai Changzheng Hospital
关键词:
自噬；肾缺血再灌注损伤；Beclin 1；LC3
Autophagy；Renal ischemia reperfusion injury；Beclin l；LC3
摘要:
目的 研究大鼠肾脏缺血再灌注（I/R）损伤后自噬溶酶体相关蛋白随损伤时间表达的规律，检测自噬体、溶酶体的变化。方法 使用肾动脉夹闭法制作大鼠肾脏I/R损伤模型，实验动物随机分入I/R组和对照组，每组6只。在肾脏缺血45 min后，分别再灌注2、6、12和24 h，使用免疫蛋白印迹法检测各时间点大鼠肾脏I/R损伤后组织内自噬溶酶体相关蛋白的表达情况，透射电镜观察肾I/R损伤后组织内自噬小体的形成。结果 I/R组肾组织内Beclin 1和LC3蛋白表达水平均自I/R后2 h起开始上调，至I/R后24 h到达高峰（P值均<0.05）。透射电镜显示，对照组大鼠近端小管上皮细胞核膜完整，染色质结构正常，胞质中细胞器形态正常，未见自噬小体；I/R组大鼠I/R后2 h可见到损伤区域自噬体形成，溶酶体增多，12～24 h最为明显。结论 大鼠肾I/R损伤后组织的自噬活性上升。
Objective To investigate the expression of autophagy/lysosome associated proteins and to detect the changes of autophagy and lysosome after renal ischemia reperfusion injury in rats．Methods Twelve rats were randomly divided into ischemia reperfusion group and control group(n=6).he renal ischemia reperfusion injury was achieved by renal artery occlusion. The expression of autophagy/lysosome associated proteins after renal ischemia reperfusion injury were determined by immunoblotting with specific antibody to Beclin 1 and LC3.The formation of autophagosome and the activation of lysosome were observed under transmission electron microscope (TEM). Results The expression of Beclin l and LC3 in nephridial tissue was increased at 2h after renal ischemia reperfusion. and peaked at 24h(all P <0.05). Autophagosomes were found and lysosomes were increased under TEM at 2 h in ischemic cortex after renal ischemia reperfusion and the number of autophagosomes and lysosomes was significantly increased at 12-24 h．While the nuclear membrane was intact and the structure of chromatin and organelles were normal in the epithelial cells of proximal tubule in the control group. Conclusion Autophagy/lysosome pathway can be activated after renal ischemia reperfusion injury in rats.

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