王影, 周峰, 徐传英, 孙红
徐州医学院 病理生理学教研室； 生理学教研室，徐州 221002
本文旨在观察转染β1- 肾上腺素受体(β1-adrenoceptor, β1-AR)基因对异丙肾上腺素(isoprenaline, ISO)损伤的大鼠心肌细胞收缩功能和存活的影响。采用胶原酶消化法分离培养原代大鼠心肌细胞，转染含β1-AR 基因的腺病毒后，用ISO 孵育24 h 致心肌细胞损伤，采用Western blot 检测心肌细胞β1-AR 蛋白的含量，通过计数培养细胞中杆状细胞的百分率来检测心肌细胞存活率，用可视化边缘探测系统检测心肌细胞收缩功能变化。结果显示，与对照组相比，转染β1-AR 基因的心肌细胞β1-AR 蛋白含量明显增加(P<0.01) ；转染 β1-AR 基因并用ISO 损伤的心肌细胞内β1-AR 蛋白含量亦明显增加(P<0.01)。与对照组相比，转染β1-AR 基因的心肌细胞存活率没有明显变化，ISO 损伤的心肌细胞存活率明显下降(P<0.01)；而转染 β1-AR基因并用ISO 损伤的心肌细胞存活率与ISO 组比进一步下降(P<0.01)。与对照组相比，转染β1-AR 基因的心肌细胞收缩幅度显著增大(P<0.05)，ISO 损伤的心肌细胞收缩幅度明显下降(P<0.01)；而转染β1-AR 基因并用ISO 损伤的心肌细胞的收缩幅度与ISO 组比没有明显变化。以上结果提示，β1-AR 的表达增加对ISO 损伤的大鼠心肌细胞的保护作用不明显。
[Overexpression of β1-adrenoceptor can not protect rat cardiomyocytes from injury induced by isoprenaline.] [Ariticle in Chinese]
WANG Ying, ZHOU Feng, XU Chuan-Ying, SUN Hong
Department of Pathophysiology； Department of Physiology, Xuzhou Medical College, Xuzhou 221002, China
The purpose of this study was to investigate the effect of the overexpression of β1-adrenoceptor (β1-AR) on the contractilefunction and cell survival of rat cardiomyocytes injured by isoprenaline (ISO). The rat cardiomyocytes were isolated using thecollagenase perfusion method and then transfected with β1-AR gene using adenoviruses vector. Four hours after the infection, the ratcardiomyocytes were treated with ISO for 24 h to imitate the high catecholamine levels of chronic heart failure. Western blot wasperformed to measure the protein expression of β1-AR. The percentages of rod cells were measured to test cell survival. Video-basededge-detection system was used to measure the contractile function of the cardiomyocytes. The results indicated that the expression ofβ1-AR in β1-AR-transfected cardiomyocytes was significantly increased compared with that in control group (P<0.01). Meanwhile, β1-ARtransfection also increased β1-AR protein levels in ISO-injured cardiomyocytes. The cardiomyocyte survival was significantly decreasedin ISO group compared with that in control group. β1-AR-transfection alone had no effect on cardiomyocyte survival in β1-ARgroup, but it further decreased cardiomyocyte survival in β1-AR+ISO group. Contractile amplitudes of ISO-injured cardiomyocyteswere significantly decreased regardless of whether they were transfected with β1-AR or not, although β1-AR-transfected cardiomyocytesshowed significantly increased contractile function compared with control group (P<0.05). These results suggest that the overexpressionof β1-AR has no significant protective effect on rat cardiomyocytes injured by ISO.
通讯作者：孙红 E-mail: firstname.lastname@example.org
王影, 周峰, 徐传英, 孙红. β1- 肾上腺素受体表达的增加未能保护异丙肾上腺素损伤的大鼠心肌细胞[J]. 生理学报 2010; 62 (6): 505-510.
WANG Ying, ZHOU Feng, XU Chuan-Ying, SUN Hong. [Overexpression of β1-adrenoceptor can not protect rat cardiomyocytes from injury induced by isoprenaline.] [Ariticle in Chinese] . Acta Physiol Sin 2010; 62 (6): 505-510 (in Chinese with English abstract).