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β(1)-肾上腺素能受体过表达不能保护大鼠心肌细胞免受异丙肾上腺素诱导的损伤

[Overexpression of β(1)-adrenoceptor can not protect rat cardiomyocytes from injury induced by isoprenaline].

作者信息

Wang Ying, Zhou Feng, Xu Chuan-Ying, Sun Hong

机构信息

Department of Pathophysiology, Xuzhou Medical College, Xuzhou 221002, China.

出版信息

Sheng Li Xue Bao. 2010 Dec 25;62(6):505-10.

Abstract

The purpose of this study was to investigate the effect of the overexpression of β(1)-adrenoceptor (β(1)-AR) on the contractile function and cell survival of rat cardiomyocytes injured by isoprenaline (ISO). The rat cardiomyocytes were isolated using the collagenase perfusion method and then transfected with β(1)-AR gene using adenoviruses vector. Four hours after the infection, the rat cardiomyocytes were treated with ISO for 24 h to imitate the high catecholamine levels of chronic heart failure. Western blot was performed to measure the protein expression of β(1)-AR. The percentages of rod cells were measured to test cell survival. Video-based edge-detection system was used to measure the contractile function of the cardiomyocytes. The results indicated that the expression of β(1)-AR in β(1)-AR-transfected cardiomyocytes was significantly increased compared with that in control group (P<0.01). Meanwhile, β(1)-AR transfection also increased β(1)-AR protein levels in ISO-injured cardiomyocytes. The cardiomyocyte survival was significantly decreased in ISO group compared with that in control group. β(1)-AR-transfection alone had no effect on cardiomyocyte survival in β(1)-AR group, but it further decreased cardiomyocyte survival in β(1)-AR+ISO group. Contractile amplitudes of ISO-injured cardiomyocytes were significantly decreased regardless of whether they were transfected with β(1)-AR or not, although β(1)-AR-transfected cardiomyocytes showed significantly increased contractile function compared with control group (P<0.05). These results suggest that the overexpression of β(1)-AR has no significant protective effect on rat cardiomyocytes injured by ISO.

摘要

本研究旨在探讨β(1)-肾上腺素能受体(β(1)-AR)过表达对异丙肾上腺素(ISO)损伤的大鼠心肌细胞收缩功能和细胞存活的影响。采用胶原酶灌注法分离大鼠心肌细胞,然后用腺病毒载体转染β(1)-AR基因。感染4小时后,用ISO处理大鼠心肌细胞24小时以模拟慢性心力衰竭时的高儿茶酚胺水平。进行蛋白质免疫印迹法检测β(1)-AR的蛋白表达。测量杆状细胞百分比以检测细胞存活情况。使用基于视频的边缘检测系统测量心肌细胞的收缩功能。结果表明,与对照组相比,β(1)-AR转染的心肌细胞中β(1)-AR的表达显著增加(P<0.01)。同时,β(1)-AR转染也增加了ISO损伤的心肌细胞中β(1)-AR蛋白水平。与对照组相比,ISO组心肌细胞存活率显著降低。单独β(1)-AR转染对β(1)-AR组心肌细胞存活无影响,但在β(1)-AR+ISO组中进一步降低了心肌细胞存活率。无论是否转染β(1)-AR,ISO损伤的心肌细胞的收缩幅度均显著降低,尽管β(1)-AR转染的心肌细胞与对照组相比收缩功能显著增强(P<0.05)。这些结果表明,β(1)-AR过表达对ISO损伤的大鼠心肌细胞无显著保护作用。

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