Integration of network pharmacology and metabolomics to elucidate Qi-Xiong-Qu-Yu decoction mitigates myocardial ischemia via regulating Nrf2/ROS/MAPK signaling

Abstract

Background: Qi-Xiong-Qu-Yu decoction (QXD) is an empirical herbal formula developed for the clinical treatment of ischemic heart disease. Methods: Isoproterenol (ISO) was used to establish a rat model of myocardial ischemia, and the effects of QXD were evaluated. The chemical profile of QXD was identified using UPLC-Q-Exactive-MS. Network pharmacology was then performed based on the UPLC-MS identified constituents and GEO datasets, and plasma metabolomics was employed to predict the molecular signaling. Finally, cell and animal experiments were conducted to verify the predicted results. Results: QXD exerted therapeutic effects in ISO-induced myocardial ischemia rats. A total of 23 constituents were identified in QXD via UPLC-MS. Integrated network pharmacology and metabolomics analyses indicated that regulation of the Nrf2/ROS/MAPK pathway may underlie the anti-myocardial ischemia effects of QXD. In vivo, QXD significantly lowered the elevated serum levels of myocardial injury biomarkers (cTnT, CK-MB, and LDH) and decreased myocardial MDA levels, attenuated cardiomyocyte apoptosis and myocardial fibrosis, and restored the activities of antioxidant enzymes including SOD and GSH-Px. In vitro, QXD reduced the release of cTnT, CK-MB, and LDH into the cell culture supernatant, decreased intracellular ROS and MDA levels, and increased SOD activity in ISO-stimulated H9C2 cells. Our study suggests that the cardioprotective effects of QXD may be mediated by promoting the nuclear accumulation of Nrf2, which upregulates the expression of antioxidant enzymes, thereby inhibiting the activation of MAPK signaling. Conclusion: QXD mitigated ISO-induced myocardial ischemia in rats, and the underlying molecular mechanisms are closely associated with the suppression of cardiomyocyte apoptosis via the Nrf2/ROS/MAPK pathway.