Qiangxin bushen decoction attenuates cardiorenal syndrome type II via AMPK/FOXO1-mediated ferroptosis pathway: A multi-omics and experimental study

BACKGROUND: Our previous studies have demonstrated that Qiangxin Bushen Decoction (QBD) exhibits therapeutic effects in heart failure patients complicated with renal insufficiency. However, the precise molecular pathways and pathophysiological mechanisms through which QBD ameliorates cardiorenal syndrome type II (CRS2) progression remain unclear. PURPOSE: To explore the mechanism of QBD for the treatment of CRS2. METHODS: UPLC-Q-TOF-MS was employed to explore comprehensive metabolite profiling from QBD samples. Network pharmacology, RNA-seq, and metabolomics were utilized to predict the bioactive constituents, potential therapeutic targets, and intervention pathways through which QBD exerts its effects on CRS2. Cardiac and renal function in mice were assessed by echocardiography along with serum and urinary biochemical markers. Cardiac and renal tissue samples were collected for histological, protein, and genetic examinations, and targeted genes werevalidated using immunohistochemistry (IHC) staining, western blotting(WB) analysis, and qPCR analysis. RESULTS: A total of 74 compounds were identified in QBD samples, and 118 compounds were detected in QBD-containing serum. Echocardiography revealed significant ventricular remodeling in CRS2 mice, which was markedly ameliorated following QBD treatment. Serum and urine biochemical assays further confirmed that QBD effectively alleviated cardiorenal injury. Histopathological examination demonstrated significantly reduced pathological damage in the heart and kidney tissues of QBD-treated mice. Multimodal validation, including IHC staining, WB, and qPCR analyses, indicated that QBD attenuated tissue ferroptosis by modulating the AMP-activated protein kinase (AMPK)/Forkhead box protein O1 (FOXO1) pathway in CRS2 mice. In vitro experiments using AMPK inhibitor Dorsomorphin and ferroptosis activator Erastin confirmed that QBD exerted its therapeutic effects via this pathway. Integrated transcriptomic and metabolomic analyses revealed that QBD treatment modulated metabolic pathways associated with energy metabolism, oxidative stress, and biosynthetic functions. CONCLUSION: This study reveals that QBQ exerts its cardiorenal protection for CRS2 through AMPK/FOXO1-dependent ferroptosis inhibition with contemporary multi-omics research tools.