Fan Xin-Yi, Wei Xiao-Qi, Chai Wang-Jing, Li Fang-He, Gao Kuo, Yu Xue, Guo Shu-Zhen
School of Traditional Chinese Medicine, Beijing University of Chinese Medicine Beijing 100029, China.
Zhongguo Zhong Yao Za Zhi. 2025 Jun;50(11):3108-3116. doi: 10.19540/j.cnki.cjcmm.20250122.402.
This study employed a mouse model of coronary artery ligation to assess the effect and mechanism of Jiming Powder on mitochondrial autophagy in mice with myocardial infarction. The mouse model of heart failure post-myocardial infarction was established by ligating the left anterior descending coronary artery. The pharmacological efficacy of Jiming Powder was evaluated through echocardiographic imaging, hematoxylin-eosin(HE) staining, and Masson staining. The levels of malondialdehyde(MDA), Fe(2+), reduced glutathione(GSH), and superoxide dismutase(SOD) in heart tissues, as well as MDA immunofluorescence of heart tissues, were measured to assess lipid peroxidation and Fe(2+) levels in the hearts of mice in different groups. Ferroptosis levels in the groups were evaluated using scanning electron microscopy and Prussian blue staining. Western blot analysis was conducted to detect the levels of key ferroptosis-related proteins, including nuclear factor erythroid 2-related factor 2(NRF2), ferritin heavy chain(FTH), glutathione peroxidase 4(GPX4), solute carrier family 7 member 11(SLC7A11), heme oxygenase 1(HO-1), and Kelch-like ECH-associated protein 1(KEAP1). The results showed that compared with the model group, both the high-and low-dose Jiming Powder groups exhibited significantly reduced left ventricular internal diameter in systole(LVIDs) and left ventricular internal diameter in diastole(LVIDd), while the left ventricular ejection fraction(EF) and left ventricular fractional shortening(FS) were significantly improved, effectively enhancing cardiac function in mice post-myocardial infarction. HE staining revealed that Jiming Powder attenuated myocardial inflammatory cell infiltration post-infarction, and Masson staining indicated that Jiming Powder effectively reduced fibrosis in the infarct margin area. Treatment with Jiming Powder reduced the levels of MDA and Fe~(2+), indicators of lipid peroxidation post-myocardial infarction, while increasing GSH and SOD levels, thus protecting ischemic myocardium. Western blot results demonstrated that Jiming Powder reduced KEAP1 protein accumulation, activated the NRF2/HO-1/GPX4 pathway, and up-regulated the protein expression of FTH and SLC7A11, exerting an inhibitory effect on ferroptosis. This study reveals that Jiming Powder exerts a therapeutic effect on myocardial infarction by inhibiting ferroptosis through the NRF2/HO-1/GPX4 pathway, providing a foundation for subsequent research on the pharmacological effects of Jiming Powder.
本研究采用冠状动脉结扎小鼠模型，以评估鸡鸣散对心肌梗死小鼠线粒体自噬的影响及机制。通过结扎左冠状动脉前降支建立心肌梗死后心力衰竭小鼠模型。通过超声心动图成像、苏木精-伊红（HE）染色和Masson染色评估鸡鸣散的药理作用。检测心脏组织中丙二醛（MDA）、Fe²⁺、还原型谷胱甘肽（GSH）和超氧化物歧化酶（SOD）水平，以及心脏组织的MDA免疫荧光，以评估不同组小鼠心脏的脂质过氧化和Fe²⁺水平。使用扫描电子显微镜和普鲁士蓝染色评估各组的铁死亡水平。进行蛋白质免疫印迹分析以检测关键铁死亡相关蛋白的水平，包括核因子红细胞2相关因子2（NRF2）、铁蛋白重链（FTH）、谷胱甘肽过氧化物酶4（GPX4）、溶质载体家族7成员11（SLC7A11）、血红素加氧酶1（HO-1）和 Kelch样ECH相关蛋白1（KEAP1）。结果表明，与模型组相比，高剂量和低剂量鸡鸣散组的收缩末期左心室内径（LVIDs）和舒张末期左心室内径（LVIDd）均显著减小，而左心室射血分数（EF）和左心室短轴缩短率（FS）显著改善，有效增强了心肌梗死后小鼠的心脏功能。HE染色显示，鸡鸣散减轻了梗死后心肌炎症细胞浸润，Masson染色表明，鸡鸣散有效减少了梗死边缘区的纤维化。鸡鸣散治疗降低了心肌梗死后脂质过氧化指标MDA和Fe²⁺的水平，同时提高了GSH和SOD水平，从而保护缺血心肌。蛋白质免疫印迹结果表明，鸡鸣散减少了KEAP1蛋白积累，激活了NRF2/HO-1/GPX4通路，并上调了FTH和SLC7A11的蛋白表达，对铁死亡发挥抑制作用。本研究表明，鸡鸣散通过NRF2/HO-1/GPX4通路抑制铁死亡，对心肌梗死发挥治疗作用，为鸡鸣散药理作用的后续研究提供了依据。
