腺苷对缺氧复氧心肌细胞的保护作用_图文(精)

研究论文

腺苷对缺氧/复氧心肌细胞的保护作用

王兴祥1,3,周利龙1,丁家望2,冯义柏1,程龙献1

1华中科技大学同济医学院附属协和医院心内科,武汉430022;2宜昌市中心人民医院,宜昌443000

摘要:本研究旨在探讨腺苷(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(110μmol/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Ca2+和丙二醛(MDA浓度。用EL ISA法检测肿瘤坏死因子(TNF2α的表达,并用凝胶电泳迁移率改变法(EMSA测定核因子(NF2κB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<0101;(3ADO降低缺氧和复氧期间心肌细胞内的Ca2+浓度(both P<0101;(4ADO降低缺氧和复氧期间心肌细胞MDA浓度(both P<0101;(5ADO抑制缺氧和复氧期间TNF2α的表达(both P<0101;(6ADO抑制缺氧和复氧期间心肌细胞NF2κB结合活性(both P<0101。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TNF2α的表达;

(3外源性ADO可能通过抑制心肌细胞NF2κB结合活性下调TNF2α的表达。

关键词:腺苷;缺氧/复氧损伤;心肌细胞

中图分类号:Q463;R540

Adenosine protects cardiomyocytes from hypoxia/reoxygenation injury

WAN G Xing2Xiang1,3,ZHOU Li2Long1,DIN G Jia2Wang2,FEN G Y

i2Bai1,CHEN G Long2Xian1 1Depart ment of Cardiology,U nion Hospital A f f iliated to Tongji Medical College,Huaz hong U niver2 sity of

Science and Technology,W uhan430022;2Yichang People Hospital,Yichang443000

Abstract:The aim of this study was to investigate the protective effect of adenosine(ADOon car2 diomyocytes following hypoxia/reoxygenation(H/Rand its molecular mechanism.Primary cultured cardiomyocytes of neonatal rats were divided

in to two groups,namely H/R(controland ADO(110μmol/Lgroups.The morphologic changes in cardiomyocytes were observed under an inverted phase2 contrast microscope.The following parameters of the two groups were determined:lactate dehydroge2 nase(LDHactivity,intracellular calcium concentration and malondialdehyde(MDAcontent.Tumor necrotic factor(TN F2αassay was performed using an EL ISA kit and N F2κB in the nucleus was analyzed by electrophoretic mobility shift assay(EMSA.The results are as follows:(1after H/R injury,car2 diomyocytes contracted,tending to get round in shape and its pseudopods decreased,while marked mor2 phological changes were not observed in ADO group;(2LDH leakage maintained at a lower level in ADO group than that in the control group during H/R(both

P<0101;(3ADO significantly reduced the concentration of calcium in cells and prevented calcium overload during H/R(both P<0101;(4 ADO markedly reduced the content of MDA during H/R(both P<0101;(5ADO inhibited the pro2 duction of TN F2αduring

H/R(both P<0101;and(6ADO down2regulated N F2κB binding activity of cardiomyocytes during H/R(both P<0101。The results suggest that(1exogenous ADO attenuates

Received2002205207Accepted2002208226

3Corresponding author.Present address:Cardiovascular Disease Department,First Affiliated Hospital,Medical School of Zhejiang University,