当归红芪超滤物对心肌细胞Hsp70表达的影响

Effect of ultra-filtration extract mixture from Angelica Sinensis and Hedysarum Polybotrys on expression of Hsp70 in cardiomyocytes

目的:探讨当归红芪超滤物对凋亡心肌细胞热休克蛋白70(Hsp70)表达的影响,阐明当归红芪超滤物对心肌细胞的抗氧化作用。方法:原代培养的Wistar乳鼠心肌细胞用高浓度的H2O2(400μmol.L-1)建立细胞凋亡模型,用不同浓度(3.75、7.50和15.00g.L-1)的当归红芪超滤物进行干预,倒置显微镜下观察各实验组心肌细胞搏动频率,流式细胞仪检测心肌细胞凋亡率,分别用RT-PCR技术和Western blotting技术检测心肌细胞中Hsp70基因及蛋白的表达情况。结果:与对照组比较,H2O2损伤组心肌细胞搏动频率显著降低(P<0.01),细胞凋亡率显著增加(P<0.01),Hsp70mRNA及蛋白表达量升高(P<0.05,P<0.01);与H2O2损伤组比较,当归红芪超滤物高、中、低剂量组心肌细胞搏动频率显著增加,但低于对照组(P<0.01),凋亡率显著降低(P<0.01),Hsp70mRNA及蛋白表达量显著升高(P<0.05,P<0.01)。结论:当归红芪超滤物具有抗氧化损伤心肌细胞凋亡的作用,且可上调心肌细胞Hsp70的表达。

Objective To investigate the effect of the ultra filtration extract mixture from Angelica Sinensis and Hedysarum Polybotrys on heat shock protein 70 （Hsp70） expression in cardiomyocytes, and clarify its antioxidation on cardiomyocytes. Methods Myocardial cells from 1- 3 d neonatal rats were cultivated in DF medium and the cellular injury was induced by H202 （400 μmol · L^- 1）, The ultra-filtration extract mixtures were given in three doses of 3.75, 7.50, and 15.00 g · L^- 1. The beating rates of cardiomyocytes were observed by phase-contrast microscope. The apoptotic rates of eardiomyocytes were measured by flow cytometry. The expressions of Hsp70 mRNA and protein in eardiomyocytes were measured by RT-PCR and Western blotting. Results Compared with normal control group, the beating rates of cardiomyocytes were significantly decreased （P〈0.01） and the apoptotic rates of eardiomyocytes were significantly increased （P〈0.01） in model group, and the expressions of Hsp70 mRNA and protein in cardiomyocytes were increased （P〈0.05, P〈0.01）. Compared with model group, the beating rates of cardiomyocytes were significantly increased （P〈0. 01） and the apoptotic rates were significantly decreased （P〈0.01） in each treatment group, and the expressions of Hsp70 mRNA and proteinin cardiomyocytes were significantly increased （P〈0.01, P〈0.05）. Conclsion The ultra-filtration extract mixture has the protective effects on cardiomyocytes injured by H2O2 and can up-regulate the Hsp70 expression in cardiomyocytes.