Attenuation of acetaldehyde-induced cell injury by overexpression of aldehyde dehydrogenase-2 (ALDH2) transgene in human cardiac myocytes: role of MAP kinase signaling
Attenuation of acetaldehyde-induced cell injury by overexpression of aldehyde dehydrogenase-2 (ALDH2) transgene in human cardiac myocytes: role of MAP kinase signaling
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DOI：
10.1016/j.yjmcc.2005.11.006
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发表时间：
2006-02-01
影响因子：
5
通讯作者：
Ren, J
中科院分区：
文献类型：
--
作者：
Li, SY;Li, Q;Ren, J
乙醛是乙醇的主要代谢物，其毒性和反应性远高于乙醇，可能与酒精引起的心脏损害有关。本研究旨在探讨转人乙醛脱氢酶-2(ALDH2)基因促进乙醛代谢对乙醛和乙醇诱导的细胞损伤的影响。将ALDH2基因导入人胎心肌细胞，通过流式细胞仪、Western印迹和ALDH2活性检测证实了ALDH2基因的导入效果。用5-(6)-chloromethyl-2‘，7’-dichlorodihydrofluorescein二乙酸酯(CM-H2DCFDA)检测ROS的产生。4‘，6’-diamidine-2‘-phenylindoladihydrochloride荧光显微镜、定量DNA片段酶联免疫吸附试验和半胱氨酸天冬氨酸氨基转移酶3活性检测细胞凋亡。乙醛和乙醇孵育24-48h后，可诱导人心肌细胞产生明显的ROS并诱导细胞凋亡。免疫组织化学染色显示乙醛处理的心肌细胞中MAP激酶ERK1/2、SAPK/JNK和p38MAPK级联激活。有趣的是，ALDH2转基因显著抑制乙醛诱导的ROS生成、细胞凋亡和ERK1/2和SAPK/JNK的磷酸化。时间依赖性反应(0~12h)显示乙醛诱导的细胞凋亡前ROS积聚和MAP激酶的激活。此外，非酶类抗氧化剂可拮抗乙醛诱导的ROS生成和细胞凋亡。我们的结果提示，ALDH2转基因过表达可能通过ERK1/2和Spak/JNK依赖机制有效地减轻乙醛诱导的细胞损伤。我们的数据与乙醛是酒精性心肌病的诱因的观点是一致的，并暗示了ALDH2酶在酒精性并发症中的治疗潜力。(C)2005爱思唯尔有限公司。保留所有权利。
Acetaldehyde, the major metabolite of ethanol, which is far more toxic and reactive than ethanol, may be responsible for alcohol-induced cardiac damage. This Study was designed to examine the impact of facilitated acetaldehyde metabolism using transfection of human aldehyde dehydrogenase-2 (ALDH2) transgene on acetalclehyde- and ethanol-induced cell injury. Fetal human cardiac myocytes were transfected with ALDH2, the efficacy of which was verified by flow cytometry, Western blot and ALDH2 activity assays. Generation of reactive oxygen species (ROS) was detected using 5-(6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H2DCFDA). Apoptosis was evaluated by 4',6'-diamidine-2'-phenylindoladihydrochloride (DAPI) fluorescence microscopy, quantitative DNA fragmentation ELISA and caspase 3 activity. Acetaldehyde and ethanol elicited overt ROS generation and apoptosis in human cardiac myocytes following 24-48 h of incubation. Immunostaining revealed activation of the MAP kinase cascades ERK1/2, SAPK/JNK and p38 MAP kinase in acetaldchyde-treated myocytes. Interestingly, ALDH2 transgene significantly attenuated acetaldehyde-induced ROS generation, apoptosis and phosphorylation of ERK1/2 and SAPK/JNK. Time-dependent response (0-12 h) revealed ROS accumulation and activation of MAP kinases prior to acetaldehyde-induced apoptosis. In addition, acetaldehyde-induced ROS generation and apoptosis were antagonized by non-enzymatic antioxidants. Our results suggested that ALDH2 transgene overexpression may effectively alleviate acetaldehyde-elicited cell injury through an ERK1/2 and SPAK/JNK-dependent mechanism. Our data are consistent with the notion of acetaldehyde as a contributor to alcoholic cardiomyopathy and implicate the therapeutic potential of ALDH2 enzyme in alcoholic complications. (c) 2005 Elsevier Ltd. All rights reserved.