过刊浏览

腺苷对缺氧/复氧心肌细胞的保护作用

王兴祥, 周利龙, 丁家望, 冯义柏, 程龙献

华中科技大学同济医学院附属协和医院心内科，武汉430022

摘要

研究旨在探讨腺苷(adenosine , ADO)对缺氧/复氧(hypoxia/ reoxygenation , H/ R)心肌细胞的保护作用及其分子机制。将原代培养的新生大鼠心肌细胞分成H/ R对照组和ADO (1 . 0 umol/ L)保护组。用倒置相差显微镜观察心肌细胞的生长状态。检测两组培养基质乳酸脱氢酶(LDH)活性和心肌细胞Ca'+和丙二醛(MDA)浓度。用ELISA法检测肿瘤坏死因子(TNF-a)的表达，并用凝胶电泳迁移率改变法(EMSA)测定核因子(NF-KB)结合活性。所得结果如下:(1)心肌细胞H/ R培养后皱缩、变圆，伪足减少，ADO组心肌细胞的形态变化小于对照组;(2)ADO减少缺氧和复氧期间心肌细胞LDH的漏出(both P<0.01) ; (3) ADO降低缺氧和复氧期间心肌细胞内的Ca~(2+) 浓度(both P<0.01) ; (4) ADO降低缺氧和复氧期间心肌细胞MDA浓度(both P<0.01) ; (5) ADO抑制缺氧和复氧期间T N F-。的表达(both P < 0. 01) ; (6) ADO抑制缺氧和复氧期间心肌细胞NF- KB结合活性(both P<0.01)。以上
结果提示:(1)外源性ADO可减轻心肌细胞的H/ R损伤;(2)外源性ADO抑制H/ R期间心肌细胞TNF-。的表达;(3)外源性ADO可能通过抑制心肌细胞NF-KB结合活性下调TNF-#alpha#的表达。

关键词： 腺苷; 缺氧/复氧损伤; 心肌细胞

Adenosine protects cardio myocytes from hypoxia/ reoxygenation injury

WANG Xinyang , ZHOU LiLong, DING Jia- Wang, FENG Yi-Bai, CHENG Long- Xian

Department of Cardiology, Union Hospital Affiliated to Tongji Medical College , Huazhong Univesity of Science and Technology, Wuhan430022； Yichang People Hospital , Yichang 443000。

Abstract

The aim of this study was to investigate the protective effect of adenosine(ADO) on car diomyocytes following hypoxia/ reoxygenation(H/ R) and its molecular mechanism.Primary cultured cardiomyocytes of neonatal rats were divided into two groups，namely H/ R(control) and ADO(1 .0p mol/ L) groups .The morphologic changes in cardiomyocytes were observed under an inverted phasecontrast microscope .The following parameters of the two groups were determined:lactate dehydrogenase(LDH) activity，intracellular calcium concentration and malondialdehyde(MDA) content .Tumor necrotic factor(TNF-a) assay was performed using an ELISA kit and NF KB in the nucleus was analyzed
by electrophoretic mobility shift assay(E MS A) .The results are as follows:(1)after H/ R injury，cardiomyocytes contracted，tending to get round in shape and its pseudopods decreased，while marked morphological changes were not observed in ADO group;(2) LDH leakage maintained at a lower level in
ADO group than that in the control group during H/ R(both P<0. 01);(3) ADO significantly reduced the concentration of calcium in cells and prevented calcium overload during H/ R(both P<0. 01);(4)ADO markedly reduced the content of MDA during H/ R(both P<0.01);(5) ADO inhibited the production of TNF-a during H/ R(both P<0. 01);and(6) ADO dower regulated NF KB binding activity of
cardiomyocytes during H/ R(both P<0.01)。The results suggest that(1)exogenous ADO attenuates H/ R injury of cultured cardiomyocytes;(2) exogenous ADO inhibits the production of TNF-a after H/ R injury;(3) exogenous ADO prevents the activation of NF-KB，which may be the molecular mechanism of wirregulation of TNF-a expression.

Key words: adenosine;hypoxia/ reoxygenation injury;cardio myocytes

收稿日期：　　录用日期：

通讯作者：　　E-mail:

引用本文：

王兴祥, 周利龙, 丁家望, 冯义柏, 程龙献. 腺苷对缺氧/复氧心肌细胞的保护作用[J]. 生理学报 2003; 55 (1): .

WANG Xinyang , ZHOU LiLong, DING Jia- Wang, FENG Yi-Bai, CHENG Long- Xian. Adenosine protects cardio myocytes from hypoxia/ reoxygenation injury. Acta Physiol Sin 2003; 55 (1): (in Chinese with English abstract).