生脉注射液抗AngⅡ诱导的心肌细胞肥大和凋亡的作用以及对能量代谢的影响

Effect of Shengmai Injection on AngⅡ-induced Cardiomyocyte Hypertrophy and Apoptosis and Its Effect on Energy Metabolism

目的研究生脉注射液对Ang Ⅱ诱导的心肌肥厚和细胞凋亡中心肌能量代谢的影响。方法使用0.8μM Ang Ⅱ诱导新生大鼠心肌细胞48 h。将心肌细胞随机分为3组:空白组(Blank),0.8μM Ang Ⅱ打击组(Ang Ⅱ)和Ang Ⅱ 0.8μM+生脉注射液稀释4000倍组(SMI)。通过透射电镜检测线粒体超微结构以评价线粒体功能,通过PCR和Western Blot检测线粒体生物发生相关基因的mRNA和蛋白的表达:AMPK、PGC-1α、CPT-1和GLUT-4。流式细胞仪检测细胞凋亡,PCR检测凋亡相关蛋白Bcl-2、Bax、caspase-3mRNA的表达。结果 (1)生脉注射液药物可以拮抗Ang Ⅱ的作用,生脉注射液稀释4000倍时拮抗效果最佳。(2)与空白组相比,Ang Ⅱ组心肌细胞活力下降,心肌细胞直径和总蛋白显著增加,线粒体超微结构受损,与能量代谢相关的基因如AMPK、PGC-1α、CPT-1和GLUT-4 mRNA和蛋白的表达下降,心肌细胞凋亡率明显下降,凋亡相关蛋白Bcl-2、Bax、caspase-3mRNA的表达(P <0.05)。与Ang Ⅱ组比较,生脉注射液组可明显逆转Ang Ⅱ对心肌细胞的影响(P <0.05)。结论生脉注射液可能通过能量依赖性机制抑制血管紧张素Ⅱ诱导的心肌细胞肥大和凋亡。

Objective To observe the effect of Shengmai Injection on myocardial energy metabolism induced by Ang Ⅱ-induced cardiac hypertrophy and apoptosis. Methods We induced neonatal rat cardiomyocytes for 48 hours using 0. 8 μM Ang Ⅱ.Cardiomyocytes were randomly divided into three groups: blank group( Blank),0. 8 μM Ang Ⅱ shock group( Ang Ⅱ) and Ang Ⅱ0. 8 μM + Shengmai Injection 1/4000 group( SMI). Mitochondrial ultrastructure was examined by transmission electron microscopy to evaluate mitochondrial function. The mRNA and protein expression of mitochondrial biogenesis related genes,such as AMPK,PGC-1α,CPT-1 and GLUT-4 were detected by PCR and Western Blot. Apoptosis was detected by flow cytometry,and the expressions of apoptosis-related proteins Bcl-2,Bax and caspase-3 mRNA were detected by PCR. Results( 1) Shengmai Injection can antagonize the effect of Ang Ⅱ,and the antagonism effect was the best when Shengmai Injection was diluted 4000 times.( 2) Compared with those of the blank group,the myocardial cell viability of the Ang Ⅱ group decreased,the myocardial cell diameter and total protein increased significantly,the mitochondrial ultrastructure was impaired,and the expressions of mRNA and protein of energy metabolism related genes such as AMPK,PGC-1α,CPT-1 and GLUT-4 decreased,the apoptosis rate of cardiomyocytes decreased significantly,and the expressions of apoptosis-related proteins Bcl-2,Bax and caspase-3 mRNA( P <0. 05). Compared with those of the Ang Ⅱ group,the Shengmai Injectionsignificantly reversed the effect of Ang Ⅱ on cardiomyocytes( P < 0. 05). Conclusion Shengmai Injection may inhibit Ang Ⅱ-induced cardiomyocyte hypertrophy and apoptosis through an energy-dependent mechanism.