天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (10): 1233-1237.doi: 10.11958/20160501

• 实验研究 • 上一篇    下一篇

缺血预适应对肝脏热缺血再灌注损伤保护作用的机制研究

金涛，胡宇，刘超   

  1. 1 天津市南开医院重症医学科（邮编 300100）；2 解放军第 452 医院麻醉科；3 天津市胸科医院心内科
  • 收稿日期:2016-06-03 修回日期:2016-08-05 出版日期:2016-10-15 发布日期:2016-10-21
  • 通讯作者: 胡宇 E-mail:2639227830@qq.com
  • 作者简介:金涛（1973）， 男， 副主任医师， 学士， 主要从事危重症医学方面研究
  • 基金资助:
    2013 年中国博士后科学基金资助项目（2013M530880）；2015 年中国博士后科学基金资助项目（2015M581308）

The pathogenesis of ischemic preconditioning to warm ischemia reperfusion injury of hepatocytes in rats

JIN Tao, HU Yu, LIU Chao   

  1. 1 Department of ICU， Tianjin Nankai Hospital, Tianjin 300100, China; 2 Department of Anesthesiology， People’ s Liberation Army No. 452 Hospital; 3 Department of Cardiology, Tianjin Chest Hospital
  • Received:2016-06-03 Revised:2016-08-05 Published:2016-10-15 Online:2016-10-21
  • Contact: HU Yu E-mail:2639227830@qq.com

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摘要： 目的 探讨缺血预适应对肝脏热缺血再灌注损伤保护作用的分子机制。 方法 90 只成年清洁级 SD 大鼠随机均分为 3 组， A 组为假手术组， B、C 组建立 SD 大鼠肝脏热缺血再灌注损伤模型， 并对 C 组大鼠进行缺血预适应干预。 于缺血再灌注后 0、2、6、12、24 h 采集血标本测定丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）， 采用酶联免疫吸附（ELISA）技术检测肝细胞中肿瘤坏死因子（TNF）-α 及白细胞介素（IL）-1β 水平； 采集肝组织标本测定丙二醛（MDA）和超氧化物歧化酶（SOD）， 流式细胞仪测定肝细胞线粒体膜电位。 结果 B、C 组大鼠的血清转氨酶、TNF-α、 IL-1β 及 MDA 水平均明显高于 A 组（P＜ 0.05）； B、C 组大鼠的凝血酶原活动度和胆碱酯酶低于 A 组（P＜ 0.05）； B、C 组大鼠肝脏 SOD 水平明显低于 A 组；C 组大鼠各项指标均优于 B 组（P＜ 0.05）。 缺血再灌注发生后肝细胞线粒体膜电位水平在 0 h 后即达到最低值， 此后呈逐渐上升趋势（P＜ 0.05）， 并且 C 组的肝细胞线粒体膜电位水平在各时段均高于 B 组（P＜ 0.05）。 结论 缺血预适应对肝细胞热缺血再灌注损伤具有一定的保护作用， 缺血预适应可以通过减少炎症因子 TNF-α 及 IL-1β 的释放， 提高 SOD 拮抗自由基的活性， 减轻线粒体损伤等途径发挥其保护作用。

关键词: 丙氨酸转氨酶, 天冬氨酸氨基转移酶类, 再灌注损伤, 氧化性应激, 线粒体, 缺血预适应

Abstract: Objective To explore the pathogenesis of ischemic preconditioning to warm ischemia reperfusion injury of hepatocytes in rats. Methods Ninety SD rats were randomly divided into three experimental groups: sham operation group （group A）, warm hepatic ischemia/reperfusion group（group B and group C）. Group C was given ischemic preconditioning treatment. Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected 0 h, 2 h, 6 h, 12 h and 24 h after ischemia reperfusion injury. Levels of TNF- α and IL- 1β were tested detected by ELISA. Levels of malondialdehyde (MDA) and superoxide dismutase (SOD) of hepatocytes were detected at the same time points. Mitochondrial membrane potential was examined to assess ischemia reperfusion injury of hepatocytes in rats using chart of intensity of JC-1 in mitochondria. Results The serum levels of ALT, TNF-α, IL-1β, and MDA were significantly higher in hepatic warm ischemia reperfusion group and ischemic preconditioning group than those in sham operation group (P＜ 0.05). Values of prothrombin activity and cholinesterase were significantly lower in liver warm ischemia reperfusion group and ischemic preconditioning group than those of sham operation group (P＜ 0.05). The SOD level of liver was significantly lower in warm ischemia reperfusion group and ischemic preconditioning group than that in sham operation group. The indexes were better in ischemic preconditioning group than those of warm ischemia reperfusion group (P＜ 0.05). The mitochondrial membrane potential level of liver cells reached the lowest value 0 hours after ischemia and reperfusion, and then increased gradually within 24 hours (P＜ 0.05). And the level of mitochondrial membrane potential of liver cells was significantly higher in ischemic preconditioning group than that in warm ischemia reperfusion group (P＜ 0.05). Conclusion Ischemic preconditioning may play a protective role in warm ischemia- reperfusion injury in rats. Ischemic preconditioning may significantly decrease the levels of ALT, AST, TNF-α, IL-1β and MDA, and increase the SOD activity in hepatocytes. The damage of mitochondrial membrane potential is decreased after ischemic preconditioning, which might be the pathogenesis of ischemic preconditioning to warm ischemia reperfusion injury of hepatocytes in rats.

Key words: alanine transaminase, aspartate aminotransferases, reperfusion injury, oxidative stress, mitochondria, ischemic preconditioning