【摘要】 目的：探讨益心附葶饮通过BNIP3/NIX信号通路调控小鼠心肌（HL-1）细胞线粒体自噬治疗心力衰竭的作用机制。方法：将30只SD大鼠随机分为正常组、益心附葶饮低剂量组（26.25 mg/kg）、益心附葶饮中剂量组（52.50 mg/kg）、益心附葶饮高剂量组（105.00 mg/kg）、曲美他嗪组（10 mg/kg），每组各6只。连续14 d灌胃给药，给药结束后，采集腹主动脉血，制备益心附葶饮含药血清。将HL-1细胞分为正常组，模型组，益心附葶饮低、中、高剂量组，曲美他嗪组，分别加入相应含药血清培养细胞。细胞计数试剂盒-8(CCK-8)法检测细胞活性；流式细胞术检测细胞活性氧（ROS）水平；蛋白质印迹（Western blotting）法检测细胞Beclin-1、Atg7、p-P62/P62、LC3、BNIP3、NIX蛋白表达水平。结果：与正常组比较，模型组细胞活性降低（P<0.01）,ROS水平升高（P<0.01）,Beclin-1、Atg7、p-P62/P62、LC3、BNIP3、NIX蛋白表达升高（P<0.01），提示细胞处于氧化应激状态，并启动了以自噬激活为主的适应性反应；与模型组比较，益心附葶饮低、中、高剂量组及曲美他嗪组细胞活性升高（P<0.05）,ROS水平下降（P<0.01）,Beclin-1、Atg7、p-P62/P62、LC3、BNIP3、NIX蛋白表达下降（P<0.05或P<0.01），益心附葶饮疗效呈现剂量-效应依赖关系，且高剂量益心附葶饮疗效不劣于曲美他嗪。结论：益心附葶饮可能通过抑制BNIP3/NIX蛋白表达，从而调控细胞线粒体自噬，进而治疗心力衰竭。更多还原

【Abstract】 Objective: To investigate the mechanism of Yixin Futing Yin in treating heart failure by regulating mitophagy in mouse cardiomyocytes(HL-1) through the BNIP3/NIX signaling pathway. Methods:Totally 30 SD rat were randomly divided into normal group, Yixin Futing Yin low-dose group(26.25 mg/kg),Yixin Futing Yin medium-dose group(52.50 mg/kg), Yixin Futing Yin high-dose group(105.00 mg/kg) and trimetazidine group(10 mg/kg), 6 mice in each group. The drug was administered by gavage for 14 consecutive days, and blood was collected from the abdominal aorta for the preparation of drug-containing serum of Yixin Futing Yin. The HL-1 cells were subdivided into normal group, model group, Yixin Futing Yin low-dose group, Yixin Futing Yin medium-dose group, Yixin Futing Yin high-dose group and trimetazidine group, and the cells were cultured by adding the drug-containing serum. The activity of the cells in each group was detected by CCK-8. The ROS level of the cells in each group was detected by flow cytometry, and Western blottingand was used to detect the protein expressions level of Beclin-1, Atg7, p-P62/P62, LC3, BNIP3 and NIX. Results: Compared with the normal group, the model group showed decreased cell viability(P <0.01),increased ROS levels(P<0.01), and elevated protein expression of Beclin-1, Atg7, p-P62/P62, LC3, BNIP3, and NIX(P<0.01), indicating that the cells were under oxidative stress and initiated an adaptive response mainly characterized by autophagy activation. Compared with the model group, the Yixin Futing Yin low-dose group,Yixin Futing Yin medium-dose group, Yixin Futing Yin high-dose group and trimetazidine group showed increased cell viability(P<0.05), decreased ROS levels(P<0.01), and reduced protein expression of Beclin-1,Atg7, p-P62/P62, LC3, BNIP3, and NIX(P <0.05 or P <0.01). The therapeutic effect of Yixin Futing Yin exhibited a dose-dependent relationship, and the high-dose Yixin Futing Yin was non-inferior to trimetazidine.Conclusion: Yixin Futing Yin may treat heart failure by regulating cellular mitophagy through inhibition of BNIP3/NIX protein expression.更多还原