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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-cadmRNA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。
This article aims to investigate the effect of adiponectin (APN) on the expression of T-cadherin (T-cad) in cardiomyocytes injured by hypoxia / reoxygenation (H / R). Primary cultured Sprague-Dawley (SD) neonatal rat cardiomyocytes were randomly divided into normal control group, H / R group, H / R + adiponectin group (3, 10, 20 and 30 μg / mL) . H / R group was cultured in hypoxia environment (high concentration of N2-saturated hypoxic solution) for 3 h, and then placed in reoxygenation environment (pure oxygen saturated reoxygenation solution) for 1 h. The APN-treated group was pre-treated with different concentrations of APN for 24 h followed by treatment with the H / R group. The release of lactate dehydrogenase (LDH) was determined by chemical colorimetry. The apoptosis of cardiomyocytes was detected by flow cytometry and TUNEL method. The expression of LDH was detected by RT-PCR and Western blotting T-cad expression was detected. The results showed that compared with the normal control group, the apoptosis rate of cardiomyocytes in H / R group was significantly increased, the release of LDH in myocardial cells was significantly increased, and the expression of T-cadherin and protein were significantly decreased; APN pretreatment could decrease the rate of cardiomyocyte apoptosis, decrease the release of LDH and up-regulate the expression of T-cad mRNA and protein in a dose-dependent manner. The above results suggest that adiponectin can reverse the H / R-induced injury and apoptosis of cardiomyocytes by up-regulating the expression of T-cad, and has a protective effect on cardiomyocytes.