■缺血性心力衰竭(HF)已成为严重危害人们生命和健康的疾病。作为一种广泛应用于临床的中药配方,新优化的生麦散(NO-SMS)已被证明对改善心脏功能显着有效,增加运动耐受力,在中国各地区的多中心临床研究中,可以减缓心力衰竭患者心肌纤维化的进展。在我们之前的药效学和毒理学研究中,我们发现,中等剂量的制剂(8.1克原药/公斤)是最有效的治疗心力衰竭,但其作用机制仍在研究中。本研究正在探索其与心肌细胞凋亡的关系。
■我们通过两部分实验进行了研究和验证,体内和体外。首先,通过结扎冠状动脉左前降支(EF≤50%)制备雄性SD大鼠心力衰竭模型,用NO-SMS配方治疗(8.1g原药/kg/d),依芬地尔(5.4mg/kg/d)或依那普利(0.9mg/kg/d)通过管饲法制备悬浮液4周。通过超声心动图评估心脏和结构的变化,H&E,和MASSON染色。Westernblot检测各组心肌细胞凋亡,qRT-PCR,和ELISA。体外细胞实验包括H2O2和NMDA分别诱导H9c2心肌细胞损伤,各组用含NO-SMS和Ifenprodil的血清孵育24h。AnnexinV-FITC/PI双染色法检测细胞凋亡,和其余的测定与体内实验一致。
■与模型组相比,NO-SMS组和依芬地尔组能够显著改善心功能,延缓心肌纤维化,减少促凋亡蛋白的表达,mRNA以及H2O2和NMDA诱导的心力衰竭大鼠和H9c2心肌细胞中Ca2+和ROS的浓度水平,能显著降低受损心肌细胞的凋亡率,有效抑制心肌细胞凋亡。
■NO-SMS配方改善心功能,抑制HF大鼠心室重构和心肌细胞凋亡,其机制可能与NMDAR信号通路的调控有关,抑制大的细胞内Ca2+向内流动,和心肌细胞中的ROS产生。
UNASSIGNED: Ischemic heart failure (HF) has become a disease that seriously endangers people\'s life and health. As a herbal formula widely used in clinical practice, new optimized Sheng-Mai-San (NO-SMS) has been shown to be significantly effective in improving cardiac function, increasing exercise tolerance, and slowing the progression of myocardial fibrosis in heart failure patients in multi-center clinical studies in various regions of China. In our previous pharmacodynamic and toxicological studies, we found that a medium-dose formulation (8.1 g of raw drug/kg) was the most effective in the treatment of heart failure, but its mechanism of action is still being investigated. The present study is exploring its relationship with cardiomyocyte apoptosis.
UNASSIGNED: We investigated and verified this through two parts of experiments, in vivo and in vitro. Firstly, we prepared male SD rats with heart failure models by ligating the left anterior descending branch of the coronary artery (EF ≤ 50%), which were treated with NO-SMS Formula (8.1 g of raw drug/kg/d), Ifenprodil (5.4 mg/kg/d) or Enalapril (0.9 mg/kg/d) prepared suspensions by gavage for 4 weeks. The cardiac and structural changes were evaluated by echocardiography, H&E, and MASSON staining. The apoptosis of cardiomyocytes in each group was detected by Western blot, qRT-PCR, and ELISA. In vitro cell experiments include H9c2 cardiomyocyte injury induced by H2O2 and NMDA respectively, and the groups were incubated with NO-SMS and Ifenprodil-containing serum for 24 h. Apoptosis was detected by Annexin V-FITC/PI double-staining method, and the rest of the assays were consistent with the in vivo experiments.
UNASSIGNED: Compared with the model group, the NO-SMS formula group and the Ifenprodil group could significantly improve cardiac function, delay myocardial fibrosis, reduce the expression of pro-apoptotic proteins, mRNA, and the concentration levels of Ca2+ and ROS in heart failure rats and H9c2 cardiomyocytes with H2O2 and NMDA-induced injury, which could significantly reduce the apoptosis rate of damaged cardiomyocytes and effectively inhibit the apoptosis of cardiomyocytes.
UNASSIGNED: NO-SMS Formula improved cardiac function, inhibited ventricular remodeling and cardiomyocyte apoptosis in HF rats, and its mechanism may be related to the regulation of the NMDAR signaling pathway, inhibition of large intracellular Ca2+ inward flow, and ROS production in cardiomyocytes.