Zhao Yu, Li Yunfei, Tong Ling, Liang Xinying, Zhang Han, Li Lan, Fan Guanwei, Wang Yi
Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.
State Key Laboratory of Core Technology in Innovative Chinese Medicine, Tasly Academy, Tasly Holding Group Co., Ltd., Tianjin, China.
Front Physiol. 2018 Feb 6;9:48. doi: 10.3389/fphys.2018.00048. eCollection 2018.
Yiqifumai Injection (YQFM) is clinically used to treat various cardiovascular diseases including chronic heart failure (CHF). The efficacy of YQFM for treating heart failure has been suggested, but the mechanism of action for pharmacological effects of YQFM is unclear. Echocardiography detection, left ventricular intubation evaluation, histopathology and immunohistochemical examination were performed in CHF rats to evaluate the cardioprotective effect of YQFM. Rat miRNA microarray and bioinformatics analysis were employed to investigate the differentially expressed microRNAs. models of AngII-induced hypertrophy and t-BHP induced oxidative stress in H9c2 myocardial cells were used to validate the anti-hypertrophy and anti-apoptosis effects of YQFM. Measurement of cell surface area, ATP content and cell viability, Real-time PCR and Western blot were performed. YQFM significantly improved the cardiac function of CHF rats by increasing left ventricular ejection fraction and fractional shortening, decreasing left ventricular internal diameter and enhancing cardiac output. Seven microRNAs which have a reversible regulation by YQFM treatment were found. Among them, miR-21-3p and miR-542-3p are related to myocardial hypertrophy and cell proliferation, respectively and were further verified by RT-PCR. Target gene network was established and potential related signaling pathways were predicted. YQFM could significantly alleviate AngII induced hypertrophy in cellular model. It also significantly increased cell viabilities and ATP content in t-BHP induced apoptotic cell model. Western blot analysis showed that YQFM could increase the phosphorylation of Akt. Our findings provided scientific evidence to uncover the mechanism of action of YQFM on miRNAs regulation against CHF by miRNA expression profile technology. The results indicated that YQFM has a potential effect on alleviate cardiac hypertrophy and apoptosis in chronic heart failure.
益气复脉注射液（YQFM）临床上用于治疗包括慢性心力衰竭（CHF）在内的各种心血管疾病。YQFM治疗心力衰竭的疗效已得到证实，但其药理作用机制尚不清楚。对CHF大鼠进行超声心动图检测、左心室插管评估、组织病理学和免疫组化检查，以评估YQFM的心脏保护作用。采用大鼠miRNA芯片和生物信息学分析来研究差异表达的微小RNA。利用AngII诱导的H9c2心肌细胞肥大模型和t-BHP诱导的氧化应激模型来验证YQFM的抗肥大和抗凋亡作用。进行细胞表面积、ATP含量和细胞活力的测定、实时PCR和蛋白质印迹分析。YQFM通过增加左心室射血分数和缩短分数、减小左心室内径和提高心输出量，显著改善了CHF大鼠的心脏功能。发现了7种受YQFM治疗可逆调节的微小RNA。其中，miR-21-3p和miR-542-3p分别与心肌肥大和细胞增殖有关，并通过RT-PCR进一步验证。建立了靶基因网络并预测了潜在的相关信号通路。YQFM可显著减轻细胞模型中AngII诱导的肥大。在t-BHP诱导的凋亡细胞模型中，它还显著增加了细胞活力和ATP含量。蛋白质印迹分析表明，YQFM可增加Akt的磷酸化。我们的研究结果为通过miRNA表达谱技术揭示YQFM对miRNA调节抗CHF的作用机制提供了科学证据。结果表明，YQFM对减轻慢性心力衰竭中的心肌肥大和细胞凋亡具有潜在作用。
