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参芪益心方对H9C2心肌细胞能量代谢的影响及机制研究
The Effect and Mechanism of Shenqiyixin Recipe on Energy Metabolism of H9C2Cardiomyocytes
【作者】 胡鸣旭;
【导师】 刘莉;
【作者基本信息】 黑龙江中医药大学 , 中西医结合临床, 2014, 博士
【摘要】 目的:观察参芪益心方对去甲肾上腺素诱导H9C2大鼠心肌细胞的保护作用,研究参芪益心方对于受损心肌细胞能量代谢的调节,探讨参芪益心方治疗心力衰竭的作用机制。方法:H9C2大鼠心肌细胞分为正常对照组、模型组、曲美他嗪组,参芪益心组。后3组分别加入正常大鼠血清10μ L、盐酸曲美他嗪含药血清10u L、参芪益心方含药血清10μL,各组均加入去甲肾上腺素(最终浓度均为2u mol/L),在加药后24小时,48小时进行指标检测及倒置显微镜观察,MTT法测定细胞存活率,闪烁荧光法检测心肌细胞脂肪酸,葡萄糖代谢率。高效液相色谱检测细胞能量合成情况,免疫印迹技术研究心肌细胞ATP合成酶F1亚单位及PPARa蛋白表达情况结果:与模型组比较,参芪益心方组和曲美他嗪组心肌存活率OD值显著升高(P<0.01),参芪益心方组显著高于曲美他嗪组(P<0.01);与模型组比较,参芪益心方组和曲美他嗪组心肌细胞脂肪酸氧化率显著下降(P<0.01),参芪益心方组显著低于曲美他嗪组(P<0.01);葡萄糖氧化率和ATP合成量显著上升(P<0.01),参芪益心方组显著高于曲美他嗪组(P<0.01);与模型组和曲美他嗪组比较,参芪益心方组心肌细胞ATP合成酶F1亚单位表达显著上升,PPARa蛋白表达显著下调。结论:1、参芪益心方对去甲肾上腺素诱导的H9C2大鼠心肌细胞具有确切的保护作用。2、参芪益心方能有效调节受损心肌细胞能量代谢,通过抑制PPARa的表达抑制脂肪酸代谢,使受损心肌细胞以葡萄糖氧化为主要能量来源,在耗氧相同的情况下高效合成ATP,维持心肌细胞代谢需要。3、参芪益心方能有效保护心肌细胞线粒体膜电位水平,调节受损心肌细胞能量代谢,这可视为参芪益心方治疗慢性心力衰竭的主要机制。
【Abstract】 Objective:To observe the protective effect of Shenqiyixin Recipe on H9C2cells induced by norepinephrine, and to investigate the Shenqiyixin Recipe’s adjustment effect on energy metabolism of H9C2cells, explore the therapeutic mechanism of Shenqiyixin Recipe on heart failure.Methods:H9C2rat cardiomyocytes were divided into normal control group, model group, Trimetazidine group and Shenqiyixin group. The last three groups were added with normal rat serum10μL, serum with10μL Trimetazidine, serum with10μL Shenqiyixin Recipe and each group added norepinephrine (final concentration was2umol/L), indicators were tested at the24h,48h. cardiomyocytes survival was detected by MTT,Metabolic rate of fatty acids and glucose were detected by blinking fluorescence,and HPLC were unutilized for detecting the ATP generation from damaged myocardial cell. immunoblotting were unutilized for detecting the expression of F1-ATPase、PPARaResults:compared with model group, Shenqiyixin Recipe group and Trimetazidine group could significantly increase OD value(P<0.01), the value of Shenqiyixin Recipe group was significantly higher than Trimetazidine group (P<0.01); compared with model group, the fatty metabolic rate of Shenqiyixin Recipe group and Trimetazidine group were significantly lower (P<0.01), and fatty metabolic rate of Shenqiyixin Recipe group was significantly lower than Trimetazidine group(P<0.01); the glucose metabolic rate of Shenqiyixin Recipe group and Trimetazidine group were significantly higher (P<0.01), and glucose metabolic rate of Shenqiyixin Recipe group was significantly higher than Trimetazidine group(P<0.01);compared with model group and Trimetazidine group. The expression of F1-ATPase in Shenqiyixin Recipe group was significantly higher and the expression of PPARa in Shenqiyixin Recipe group is significantly lower. Conclusion:1. Shenqiyixin Recipe could effectively protect the H9C2cells induced by norepinephrine.2. Shenqiyixin Recipe could effectively adjust the energy metabolism of damaged cardiomyocytes. by inhibiting the expression of PPARa which down regulating the fatty acid metabolism, the damaged cardiomyocytes could generate ATP efficiently by the glucose metabolic way to maintain cardiomyocytes metabolism.3. Shenqiyixin Recipe could effectively protect the cardiomyocytes’MMP and adjust the energy metabolism of damaged cardiomyocytes. It may be the mechanism for Shenqiyixin Recipe treating heart failure
【Key words】 H9C2Cardiomyocytes; Protect; Shenqiyixin Recipe; EnergyMetabolism;