心复康口服液干预心力衰竭大鼠心肌线粒体蛋白质组学研究

Study on Myocardial Infarction of Xinfukang Oral Liquid on Myocardial Mitochondrial Proteomics in Rats with Heart Failure

目的:研究心复康口服液对心力衰竭大鼠心肌线粒体蛋白质组学的影响,探讨其可能的分子作用机制。方法:SD大鼠随机分为造模组与假手术组,造模组采用结扎左冠状动脉前降支致心肌梗死后心力衰竭大鼠模型,术后随机分为模型组、心复康口服液组,各组连续灌胃给药8周后采集大鼠心肌,采用双向凝胶电泳和基质辅助激光解析电离飞行时间质谱技术,分析比较各组大鼠心肌线粒体蛋白质组学表达的差异,并运用蛋白免疫印迹技术对差异表达蛋白进行回复性验证。结果:模型组与假手术组相比有差异而心复康口服液可使差异趋势逆转的蛋白点为20个,其中13个表达上调,7个表达下调,经质谱分析成功鉴定出13个蛋白,主要涉及能量代谢、应激反应、氧化损伤、细胞骨架、细胞分化增殖等功能。Western blot结果显示模型组Stress-70、Nucleophosmin表达上调,ATP-α表达下调,心复康口服液组Stress-70、Nucleophosmin表达下调,ATP-α表达上调,与蛋白质组学结果一致。结论:心复康口服液可在一定程度上调节心衰大鼠心肌线粒体蛋白质组学变化,其干预机制可能与改善能量代谢、减轻应激反应、抗氧化损伤及调节细胞分化增殖等功能相关。采用双向凝胶电泳和基质辅助激光解析电离飞行时间质谱技术,以心肌线粒体为切入点,研究心复康口服液对心衰大鼠心肌线粒体蛋白质组学的影响,结果真实可靠。

This paper aimed at investigating the effects of Xinfukang Oral Liquid on mitochondrial proteomic alterations in rats with heart failure after myocardial infarction and exploring its possible mechanisms. Heart failure models were established by ligating the left anterior descending coronary artery of SD rats. Rats were randomly divided into sham group, model group, Xinfukang group. 8 weeks after drug intervention, the mitochondrial proteomic alterations of myocardial tissue were detected by two-dimensional gel electrophoresis(2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS). Furthermore, expression levels of part of the differently expressed proteins were verified by western blot. Compared with model group, 20 differentially expressed protein spots were detected in Xinfukang group, 13 of which showed increased protein expression and 7 decreased;13 differentially expressed protein spots were successfully identified by MALDI-TOF-MS. Bioinformatics analysis showed that these differential proteins were mainly associated with energy metabolism, stress reaction, oxidative damage, cyto-skeleton,cell differentiation and proliferation. Western blot results showed that the expression of Stress-70 protein and Nucleophosmin increased and the expression of ATP-αdecreased in model group. The expression of Stress-70 protein and Nucleophosmin decreased and the expression of ATP-αincreased in Xinfukang group, which shows the same results in proteomics. Xinfukang Oral Liquid can partly adjust proteomic alterations of myocardial mitochondrial in HF rats,and its intervention mechanism may involve improving energy metabolism, reliving stress reaction and oxidative damage,as well as regulating cell differentiation and proliferation. The results of comparative proteomic analysis performed by using2-DE and MALDI-TOF-MS are acurate, stable and reliable.