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目的观察β2肾上腺素能受体(β2-AR)过表达对成年大鼠心肌细胞在不同浓度的异丙肾上腺素(isoproter-enol,ISO)刺激下的保护作用及机制。方法分离培养成年大鼠心肌细胞,随机分为3组:正常组、转染EGFP基因的腺病毒组和转染人β2-AR-EGFP基因的腺病毒组。心肌细胞培养48 h后,用异丙肾上腺素在不同剂量(10-6mol/L,10-5mol/L,10-4mol/L)下刺激2 h,观察细胞在刺激前后的存活率的变化。采用Western blot检测心肌细胞β2-AR的表达。结果Western blot结果显示转染β2-AR-EGFP基因的腺病毒组β2-AR蛋白的表达量明显高于其余两组(P<0.05),在10-6mol/L,10-5mol/L浓度的ISO刺激下3组细胞的存活率校正值无显著性差异,在高浓度(10-4mol/L)ISO刺激下β2-AR过表达组细胞存活率校正值明显高于其余两组(P<0.05)。结论β2-AR过表达对高浓度(10-4mol/L)ISO刺激的成年大鼠心肌细胞的死亡有保护作用,其可能的机制是通过β2-AR与Gi蛋白的耦联作用。
Objective To observe the protective effect and mechanism of β2-adrenoceptor (β2-AR) overexpression on cardiomyocytes of adult rats stimulated with different concentrations of isoproter-enol (ISO). Methods Adult rat cardiomyocytes were isolated and cultured and divided into 3 groups at random: normal group, adenovirus transfected with EGFP gene and adenovirus transfected with human β2-AR-EGFP gene. After cultured 48 h, the isoproterenol was stimulated with isoproterenol at different doses (10-6mol / L, 10-5mol / L, 10-4mol / L) for 2 h, and the changes of cell viability before and after stimulation were observed. Western blot was used to detect the expression of β2-AR in cardiomyocytes. Results The results of Western blot showed that the expression of β2-AR protein in adenovirus group transfected with β2-AR-EGFP gene was significantly higher than that in the other two groups (P <0.05). At the concentrations of 10-6mol / L and 10-5mol / L There was no significant difference in the survival rate of three groups under ISO stimulation. The cell viability of β2-AR over-expression group was significantly higher than that of the other two groups (P <0.05 ). Conclusion The overexpression of β2-AR has a protective effect on the death of adult rat cardiac myocytes stimulated with high concentration (10-4mol / L) of ISO. The possible mechanism is through the coupling of β2-AR with Gi protein.