Abstract：
Huangqi Guizhi Wuwu Decoction (HQGZWWD) has shown promising potential in treating various cardiovascular diseases. This study aimed to elucidate the molecular basis and therapeutic role of HQGZWWD in the treatment of doxorubicin (DOX)-induced myocardial injury. The HPLC fingerprint of HQGZWWD was used to analyze the active components. A DOX-induced myocardial damage rat model was developed, and the therapeutic effects of HQGZWWD were evaluated using echocardiography, myocardial enzyme levels, and hematoxylin and eosin staining. Network pharmacology was used to screen treatment targets, and western blotting and immunohistochemistry were performed to assess cellular pyroptosis levels. Oxidative stress levels were measured using assay kits, and mitochondrial damage was examined using transmission electron microscopy. An in vitro model of DOX-induced cell damage was established, and treatment was administered using serum containing HQGZWWD and N-acetylcysteine (NAC). Oxidative stress levels were detected using assay kits and DCFH-DA, whereas cellular pyroptosis levels were assessed through WB, immunofluorescence, and ELISA assays. HQGZWWD ameliorated DOX-induced myocardial injury. Network pharmacology identified IL-1β and IL-18 as crucial targets. HQGZWWD downregulated the protein levels of the inflammatory factors IL-1β and IL-18, inhibited the expression of GSDMD-NT, and simultaneously suppressed the synthesis of Caspase-1, ASC, NLRP3, and Caspase-11. Additionally, HQGZWWD inhibited oxidative stress, and the use of NAC as an oxidative stress inhibitor resulted in significant inhibition of the GSDMD-NT protein in H9C2 cells. These findings highlight the myocardial protective effects of HQGZWWD by inhibiting oxidative stress and suppressing both canonical and non-canonical pyroptotic pathways.
摘要：
黄芪桂枝五物汤（HQGZWWD）在治疗多种心血管疾病方面显示出广阔的潜力。本研究旨在阐明HQGZWWD治疗多柔比星（DOX）诱导的心肌损伤的分子基础和治疗作用。利用HQGZWWD的HPLC指纹图谱对活性成分进行分析。建立了DOX诱导的大鼠心肌损伤模型,使用超声心动图评价HQGZWWD的治疗效果，心肌酶水平，苏木精和伊红染色。网络药理学用于筛选治疗目标，进行蛋白质印迹和免疫组织化学以评估细胞焦亡水平。使用测定试剂盒测量氧化应激水平，使用透射电子显微镜检查线粒体损伤。建立了DOX诱导细胞损伤的体外模型,使用含有HQGZWWD和N-乙酰半胱氨酸(NAC)的血清进行治疗。使用测定试剂盒和DCFH-DA检测氧化应激水平，而细胞焦亡水平通过WB评估，免疫荧光，和ELISA测定。HQGZWWD改善DOX诱导的心肌损伤。网络药理学将IL-1β和IL-18确定为关键靶标。HQGZWWD下调炎症因子IL-1β和IL-18的蛋白水平，抑制GSDMD-NT的表达，同时抑制了Caspase-1，ASC，NLRP3和Caspase-11。此外,HQGZWWD抑制氧化应激,并且使用NAC作为氧化应激抑制剂导致H9C2细胞中GSDMD-NT蛋白的显着抑制。这些发现强调了HQGZWWD通过抑制氧化应激和抑制规范和非规范的焦变途径的心肌保护作用。
