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缺氧后处理诱导的低氧诱导因子-1α 表达上调减轻H9c2心肌细胞缺氧/复氧损伤

赵焕新, 李晓宇, 姚红, 武烨, 杨蓉, 赵荣瑞, 刘慧荣

山西中医学院生理教研室，太原 030024；山西医科大学生理教研室，太原 030001；首都医科大学生理学与病理学系，北京 100069

摘要

探讨缺氧后处理(hypoxic postconditioning, PostC)对缺氧/复氧(hypoxia/reoxygenation, H/R)所致的心肌细胞的损伤以及低氧诱导因子-1α (hypoxia inducible factor-1α , HIF-1α )表达的影响，并分析二者之间可能的关系。利用H9c2心肌细胞株建立缺氧/复氧和缺氧后处理模型，通过测定细胞存活率、细胞培养液中乳酸脱氢酶(lactate dehydrogenase, LDH)的活性及caspase-3活性来反映缺氧/复氧造成的H9c2细胞的损伤，用Western blot检测H9c2细胞内HIF-1α 的蛋白水平，用real time-PCR检测细胞内HIF-1α的mRNA水平。结果显示，缺氧后处理提高了缺氧/复氧H9c2细胞的存活率，降低了LDH活性及caspase-3活性。同时，缺氧后处理增加了H9c2细胞内HIF-1α的蛋白水平。预先利用HIF-1α脯氨酸羟化酶抑制剂DMOG上调HIF-1α的蛋白水平后，由缺氧/复氧导致的H9c2细胞的损伤明显减轻，其效应与缺氧后处理完全一致。对H9c2细胞内HIF-1α蛋白水平与细胞存活率进行相关性分析，结果显示二者呈显著正相关(r=0.743, P<0.01)；而运用siRNA方法抑制细胞内HIF-1α基因表达后，显著削弱了缺氧后处理减轻缺氧/复氧细胞损伤的效应。以上结果提示，HIF-1α表达上调是缺氧后处理减轻细胞缺氧/复氧损伤的机制之一。

关键词： 缺氧后处理; 低氧诱导因子-1α; 缺氧/复氧

分类号：R331

[The up-regulation of hypoxia inducible factor-1α induced by hypoxic postconditioning protected Heart-derived H9c2 Cells against Hypoxia/Reoxygenation.] [Article in Chinese]

ZHAO Huan-Xin, LI Xiao-Yu, YAO Hong, WU Ye, YANG Rong, ZHAO Rong-Rui, LIU Hui-Rong

Department of Physiology, Shanxi University of Traditional Chinese Medicine , Taiyuan 030024, China； Department of Physiology, Shanxi Medical University, Taiyuan 030001, China； School of Basic Medical Sciences, and Cardiovascular Research Institute, Capital Medical University, Beijing 100069, China

Abstract

This study was to explore the effects of hypoxic postconditioning (PostC) on heart-derived H9c2 cells injury induced by hypoxia/reoxygenation (H/R) and the expression of hypoxia inducible factor-1α (HIF-1α), and to analyze the relationship between them. Cultured H9c2 cardiac muscle cells were used to encounter 3 h hypoxia and 2 h reoxygenation to simulate ischemia and reperfusion, or underwent 3 cycles of 5 min reoxygenation and 5 min hypoxia preceding the long reoxygenation to simulate ischemic postconditioning. Cell viability, lactic dehydrogenase (LDH) activity, and caspase-3 activity were detected respectively to image the cell injury induced by H/R. The level of HIF-1α mRNA was measured by real time-PCR. Western blot was used to determine HIF-1α protein level. The results showed that postconditioning significantly increased H9c2 cell viability, reduced the activity of LDH and caspase-3. Simultaneously, postconditioning up-regulated HIF-1α protein level. Moreover, after DMOG, an inhibitor of proline hydroxylase (PHD) which is targeted to HIF-1αdegradation, was used to stabilize HIF-1α protein level, the reduction of H9c2 cells injury was comparable to that by postconditioning. There was a significant linear positive relationship between HIF-1α protein level and cell viability(r=0.743, P<0.01). After being infected with siRNA to silence HIF-1α gene, the cardioprotective effects of postconditioning was significantly weakened. These data suggested that up-regulation of HIF-1α induced by postconditioning play an important role in the cardioprotection of postconditioning.

Key words: hypoxic postconditioning; hypoxia inducible factor-1α; hypoxia/reoxygenation

收稿日期：2012-12-21　　录用日期：2013-04-23

通讯作者：赵焕新　　E-mail: zhhx5@yahoo.com.cn

引用本文：

赵焕新, 李晓宇, 姚红, 武烨, 杨蓉, 赵荣瑞, 刘慧荣. 缺氧后处理诱导的低氧诱导因子-1α 表达上调减轻H9c2心肌细胞缺氧/复氧损伤[J]. 生理学报 2013; 65 (3): 293-300.

ZHAO Huan-Xin, LI Xiao-Yu, YAO Hong, WU Ye, YANG Rong, ZHAO Rong-Rui, LIU Hui-Rong. [The up-regulation of hypoxia inducible factor-1α induced by hypoxic postconditioning protected Heart-derived H9c2 Cells against Hypoxia/Reoxygenation.] [Article in Chinese]. Acta Physiol Sin 2013; 65 (3): 293-300 (in Chinese with English abstract).