Supplementary file 3_Poge heart-saving decoction meliorates heart failure by suppressing apoptosis and fibrosis via regulation of the PI3K/AKT pathway.xls

BackgroundPoge Heart-Saving Decoction (PHSD) is a traditional Chinese medicine formulation that has been used clinically for decades in the treatment of heart failure (HF). However, its precise therapeutic mechanisms remain incompletely understood.
MethodsThe metabolites of PHSD were characterized using UHPLC-MS/MS. Network analysis was subsequently employed to identify the mechanism. A mouse model of HF was established through intraperitoneal injection of isoproterenol (ISO), followed by treatment with PHSD at low, medium or high doses. Cardiac function parameters were evaluated by echocardiography, and NT-proBNP levels were measured. Histopathological examination of myocardial tissue was conducted, complemented by analyses of protein and mRNA expression levels related to apoptosis and fibrosis targeting the PI3K/AKT pathway.
ResultsA total of 133 metabolites in PHSD were identified. Network analysis suggested that PHSD may ameliorate HF by targeting key proteins such as AKT1, TNF, and BCL-2. In vivo experiments demonstrated that PHSD alleviated ISO-induced myocardial apoptosis by balanced BAX and BCL-2. Furthermore, PHSD significantly reduced the deposition of Collagen I and Collagen III and markedly downregulated the expression of PI3K and AKT.
ConclusionOur study demonstrated that PHSD ameliorates HF by suppressing myocardial apoptosis and fibrosis through inhibition of the PI3K/AKT pathway. These findings indicate that PHSD is a prospective therapeutic agent against HF.

背景 救心汤（PHSD）是一种已在临床应用数十年的中药方剂，用于心力衰竭（HF）的治疗，但其确切的治疗机制尚未完全阐明。
方法 采用超高效液相色谱-串联质谱（UHPLC-MS/MS）对救心汤的代谢物进行定性表征；随后通过网络分析解析其潜在作用机制。通过腹腔注射异丙肾上腺素（isoproterenol, ISO）构建心力衰竭小鼠模型，随后分别给予低、中、高剂量的救心汤干预。采用超声心动图评估小鼠心功能参数，检测N末端B型利钠肽原（NT-proBNP）水平；对心肌组织进行组织病理学检查，并针对PI3K/AKT通路相关的细胞凋亡与纤维化指标，开展蛋白质与mRNA表达水平分析。
结果 本研究共鉴定出救心汤中的133种代谢物。网络分析结果显示，救心汤可能通过靶向AKT1、TNF、BCL-2等关键蛋白发挥改善心力衰竭的作用。体内实验表明，救心汤可通过平衡BAX与BCL-2的表达，缓解异丙肾上腺素诱导的心肌细胞凋亡；此外，救心汤可显著降低Ⅰ型胶原（Collagen I）与Ⅲ型胶原（Collagen III）的沉积，并显著下调PI3K与AKT的表达水平。
结论 本研究证实，救心汤可通过抑制PI3K/AKT通路，减少心肌细胞凋亡与纤维化，从而改善心力衰竭。上述研究结果表明，救心汤是一种极具潜力的心力衰竭治疗候选药物。