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细胞色素c 在后处理抗大鼠肠缺血- 再灌注损伤细胞凋亡中的变化

褚薇薇^*, 聂蕾, 和新盈, 闫爱丽, 周伊, 吴庚利, 王殿华1

西安医学院基础部病理教研室，西安 710021；昆明医学院天然药物药理重点实验室，昆明 650031

摘要

本文旨在研究细胞色素c 在后处理抗大鼠肠缺血- 再灌注损伤细胞凋亡中的变化。将Sprague-Dawley 大鼠32 只随机分为4 组(n=8)：假手术(Sham)组、缺血- 再灌注(I/R)组、缺血预处理(IPC)组、缺血后处理(IPOST)组。应用激光共聚焦扫描显微镜检测各组大鼠肠黏膜细胞线粒体跨膜电位的变化。用Western blot 方法检测肠黏膜细胞线粒体内细胞色素c 及caspase-3 表达的变化。末端脱氧核苷酸转移酶介导的dUTP 缺口末端标记法(TUNEL)和DNA琼脂糖凝胶电泳方法检测大鼠肠黏膜细胞凋亡发生情况。实验结果显示，与缺血- 再灌注组相比，缺血后处理组大鼠肠黏膜细胞线粒体跨膜电位显著升高(P<0.05)，线粒体内细胞色素c 蛋白表达水平显著增加(P<0.05)，caspase-3 蛋白表达降低(P<0.05)，细胞凋亡率明显降低(P<0.05)。缺血后处理组与缺血预处理组相比各项指标差异无统计学意义(P>0.05)。上述结果提示缺血后处理可通过阻止线粒体释放细胞色素c 抑制凋亡发生，减轻大鼠肠缺血- 再灌注损伤。

关键词： 细胞色素c; 再灌注损伤; 细胞凋亡

分类号：R657.3

[Change of cytochrome c in postconditioning attenuating ischemia-reperfusioninduced mucosal apoptosis in rat intestine.] [Ariticle in Chinese]

CHU Wei-Wei^*, NIE Lei, HE Xin-Ying, YAN Ai-Li, ZHOU Yi, WU Geng-Li, WANG Dian-Hua

Department of Pathology, Xi’an Medical College, Xi’an 710021, China； Yunnan Pharmacological Laboratory of Natural Products, Kunming Medical College, Kunming 650031, China

Abstract

The present study aimed to investigate the change of cytochrome c in postconditioning-attenuated ischemia-reperfusion(I/R)-induced mucosal apoptosis in rat intestine compared with ischemic preconditioning (IPC). Using rat model of intestine I/R injury,male Sprague-Dawley rats weighing 220-250 g were divided into 4 groups which were Sham operation group, I/R group, IPC group andischemic postconditioning (IPOST) group. In these groups, I/R procedure was performed by the occlusion of the superior mesentericartery (SMA) for 45 min followed by reperfusion for 1 h. In Sham group, there was no intervention. In IPC group, SMA was occludedfor 5 min and reperfused for 5 min, for two cycles, before the prolonged occlusion. In IPOST group, three cycles of 30-s reperfusionand 30-s reocclusion were preceded at the start of reperfusion. After the reperfusion, the small intestines were sampled for experimentaldetection. Intestinal mucosal mitochondrial membrane potential was detected by confocal laser scanning microscopy. Expressions ofcytochrome c and caspase-3 proteins were detected using Western-blot method. The apoptosis of intestinal mucosal cells was determinedwith agarose gel electrophoresis and deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling (TUNEL) technique.Compared with I/R group, the mitochondrial membrane potentials and the expressions of cytochrome c protein were significantlyincreased, while the expressions of caspase-3 and the apoptotic rates were decreased in IPOST and IPC groups (P<0.05). There wereno significant differences between IPOST and IPC groups (P>0.05). These data provide substantial evidence that IPOST attenuates I/R-induced mucosal apoptosis by reducing the release of cytochrome c from mitochondria in the rat small intestine.

Key words: cytochrome c; reperfusion injury; apoptosis

收稿日期：2009-11-16　　录用日期：2010-01-15

通讯作者：褚薇薇　　E-mail: shch_ww@163.com

引用本文：

褚薇薇, 聂蕾, 和新盈, 闫爱丽, 周伊, 吴庚利, 王殿华1. 细胞色素c 在后处理抗大鼠肠缺血- 再灌注损伤细胞凋亡中的变化[J]. 生理学报 2010; 62 (2): 143-148.

CHU Wei-Wei, NIE Lei, HE Xin-Ying, YAN Ai-Li, ZHOU Yi, WU Geng-Li, WANG Dian-Hua. [Change of cytochrome c in postconditioning attenuating ischemia-reperfusioninduced mucosal apoptosis in rat intestine.] [Ariticle in Chinese] . Acta Physiol Sin 2010; 62 (2): 143-148 (in Chinese with English abstract).