Network Pharmacology and Untargeted Metabolomics Analysis of the Protective Mechanisms of Total Flavonoids from Chuju in Myocardial Ischemia-Reperfusion Injury

Background/Objectives: Myocardial ischemia–reperfusion injury (MIRI) is characterized by the exacerbation of tissue damage following the restoration of blood flow to the myocardium. Chuju, recognized for its homology of food and medicine, is derived from the dried capitulum of Dendranthema morifolium (Ramat.), cultivated in Chuzhou, Anhui Province, China. Our previous studies have demonstrated that the total flavonoids extracted from Chuju (TFCJ) exhibit pharmacological efficacy against MIRI. This study will further elucidate its protective mechanism. Methods: We employed an integrative approach combining untargeted metabolomics, network pharmacology, molecular docking, and in vitro experiments to elucidate the mechanistic basis of TFCJ’s protective effects against MIRI. Results: TFCJ protected H9c2 cardiomyocytes from hypoxia/reoxygenation-induced oxidative stress and apoptosis. Integrated analyses identified Nrf2 as a central regulatory node activated by AKT signaling, which, in turn, modulates antioxidant protein expression and glutathione metabolism. Further in vitro experiments demonstrated that TFCJ induced AKT phosphorylation, thereby promoting Nrf2 activation and upregulating HO-1 expression, along with genes involved in glutathione synthesis. Conclusions: TFCJ exerts cardioprotective effects by activating the AKT-Nrf2 signaling pathway, regulating the expression of antioxidant and anti-apoptotic genes, and coordinating downstream glutathione metabolism, ultimately maintaining the oxidative-apoptotic balance in myocardial cells.