过刊浏览

脂联素上调心肌细胞缺氧/复氧损伤后T-钙黏蛋白的表达

王敏, 柴颖儒, 肖传实, 赵旭静, 卫娜, 白瑞, 边云飞^*

山西医科大学第二医院心内科，太原 030001

摘要

本文旨在研究脂联素(adiponectin, APN)对缺氧/复氧(hypoxia/reoxygenation, H/R)损伤的心肌细胞T-钙黏蛋白(T-cadherin, T-cad)表达的影响。采用酶消化法原代培养Sprague-Dawley (SD)乳鼠心肌细胞，随机分为：正常对照组、H/R组、H/R+脂联素(3, 10, 20, 30 μg/mL)组。H/R组置缺氧环境(高浓度N2饱和过的缺氧液)中培养3 h后, 再置于复氧环境(纯氧饱和过的复氧液)中培养1 h。APN处理组先用不同浓度APN预处理24 h后中随后处理同H/R组。用化学比色法测定乳酸脱氢酶(LDH)的释放，通过流式细胞术和脱氧核糖核苷酸缺口末端标记(TUNEL)法来检测心肌细胞的凋亡，用RT-PCR和Western blotting方法检测T-cad的表达。结果显示，与正常对照组相比，H/R组心肌细胞凋亡率显著升高，心肌细胞LDH的释放量明显增加，T-cad mRNA和蛋白水平表达均明显下降；而和H/R组相比，APN预处理可剂量依赖性地降低心肌细胞凋亡率，减少LDH释放量，并上调T-cad mRNA和蛋白水平。以上结果提示，脂联素可能通过上调T-cad的表达逆转H/R所致的心肌细胞损伤及凋亡，对心肌细胞有保护作用。

关键词： 脂联素; 心肌细胞; 缺氧/复氧; T-钙黏蛋白

分类号：R363

[Adiponectin up-regulates the expression of T-cadherin in cardiomyocytes injured by hypoxia/reoxygenation.] [Article in Chinese]

Wang Min, CHAI Ying-Ru, XIAO Chuan-Shi, ZHAO Xu-Jing, WEI Na, BAI Rui, BIAN Yun-Fei^*

Department of Cardiology, the Second Affiliated Hospital of Shanxi Medical University, Taiyuan 030001, China

Abstract

The aim of the present study was to investigate the effects of adiponectin (APN) on the expression of T-cadherin in cultured Sprague-Dawley (SD) rat cardiomyocytes injured by hypoxia/reoxygenation (H/R). Primary myocardial cells from neonatal rats were obtained by enzymatic digestion. The cells were divided into control group, H/R group and H/R+APN (3, 10, 20 and 30 μg/mL) groups. The H/R group was incubated in anoxic environment (anoxic solution saturated with high concentration N2) for 3 h, and then in the reoxygenation environment (the reoxygenation solution saturated with pure oxygen) for 1 h. The H/R+APN group was pretreated with different concentrations of APN for 24 h prior to the initiation of H/R. The content of lactate dehydrogenase (LDH) was measured by chemistry chromatometry. Cellular apoptosis was analyzed by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The expression of T-cadherin was detected by RT-PCR and Western blotting. The results showed that, compared with control group, the apoptotic rate and release of LDH were significantly increased in the H/R group, whereas the expressions of T-cad mRNA and protein were decreased. Pretreating with APN significantly and dose-dependently decreased apoptotic rate and LDH release, and up-regulated T-cad mRNA and protein level in rat neonatal cardiomyocytes under H/R conditions. These results suggest that APN may protect cardiomyocytes against H/R-induced injury by up-regulating H/R-decreased T-cad expression.

Key words: adiponectin; cardiomyocytes; hypoxia/reoxygenation; T-cadherin

收稿日期：2011-12-06　　录用日期：2012-04-26

通讯作者：边云飞　　E-mail:

引用本文：

王敏, 柴颖儒, 肖传实, 赵旭静, 卫娜, 白瑞, 边云飞. 脂联素上调心肌细胞缺氧/复氧损伤后T-钙黏蛋白的表达[J]. 生理学报 2012; 64 (3): 296-302.

Wang Min, CHAI Ying-Ru, XIAO Chuan-Shi, ZHAO Xu-Jing, WEI Na, BAI Rui, BIAN Yun-Fei. [Adiponectin up-regulates the expression of T-cadherin in cardiomyocytes injured by hypoxia/reoxygenation.] [Article in Chinese]. Acta Physiol Sin 2012; 64 (3): 296-302 (in Chinese with English abstract).