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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24h致心肌细胞损伤,采用Westernblot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。
The aim of this study was to investigate the effects of transfection of β1-adrenoceptor (β1-AR) gene on the contractile function and survival of rat cardiomyocytes injured by isoprenaline (ISO). The primary rat cardiomyocytes were isolated and cultured by collagenase digestion. After transfection of adenovirus containing β1-AR gene, myocardial injury was induced by ISO at 24h. The content of β1-AR protein in cardiomyocytes was detected by Western blot. The percentage of cells in the rod-shaped cells to detect myocardial cell viability, using a visual edge detection system to detect myocardial contractile function changes. The results showed that compared with the control group, the content of β1-AR protein in cardiomyocytes transfected with β1-AR gene was significantly increased (P <0.01). The content of β1-AR protein in myocardial cells transfected with β1-AR gene Significantly increased (P <0.01). Compared with the control group, the survival rate of cardiomyocytes transfected with β1-AR gene did not change significantly, and the survival rate of myocardial cells damaged by ISO significantly decreased (P <0.01); while the cells transfected with β1-AR gene and survived with ISO damaged cardiomyocytes Rates further decreased with the ISO group (P <0.01). Compared with the control group, the contraction amplitude of cardiomyocytes transfected with β1-AR gene was significantly increased (P <0.05), and the contraction amplitude of myocardial cells damaged by ISO significantly decreased (P <0.01) Injured cardiomyocytes contraction amplitude and ISO group did not change significantly. The above results suggest that the increased expression of β1-AR has no obvious protective effect on myocardial cells injured by ISO.