Investigating the Mechanism of Luteolin in Ameliorating Myocardial Injury Based on GEO Chip, Network Pharmacology, Mendelian Randomization and Experimental Validation

OBJECTIVE  To explore the mechanism of luteolin amelioration of myocardial injury based on bioinformatics, network pharmacology, molecular docking, in vitro experiments, and Mendelian randomization.

METHODS  Bioinformatics and network pharmacology analysis analyze key targets and signaling pathways of luteolin in ameliorating myocardial injury. Molecular docking of luteolin to key targets was performed. At the cellular level, a cardiomyocyte injury model was established. Cell activity was assessed using the CCK-8 assay, while the mRNA expression levels of BNP, key targets, and inflammatory cytokines(IL-18, IL-1β, IL-6) were measured by qRT-PCR. Additionally, flow cytometry was employed to analyze the apoptosis rate and ROS levels, and Western blotting was used to determine the expression of apoptosis-related proteins and key components of the SRC/PI3K/AKT signaling pathway. Furthermore, the causality between key targets and myocardial injury was explored using Mendelian randomization analysis.

RESULTS  Sixty differentially expressed genes of luteolin in myocardial injury were obtained. GO and KEGG enrichment analysis indicated that luteolin exerted its biological functions primarily via the PI3K/AKT pathway and core targets(such as SRC, AKT1, ESR1, PTGS2, AR and KDR), involving mechanisms such as protein phosphorylation and response to oxidative stress. Molecular docking verified the better binding activity of luteolin to the key targets. The cellular experimental results demonstrated that luteolin effectively alleviated cardiomyocyte injury by reducing oxidative stress and the apoptosis rate, as well as by inhibiting the secretion of inflammatory cytokines and the phosphorylation of proteins associated with the SRC/PI3K/AKT pathway. Separately, Mendelian randomization analysis indicated that the SRC gene(IVW: OR=1.050; 95%CI: 1.011–1.091; P=0.011) may be a significant biomarker for the progression of myocardial injury, suggesting a potential causal relationship with the disease risk.

CONCLUSION  The protective effect of luteolin on myocardial injury is mediated by regulating the SRC/PI3K/AKT signaling pathway, and this finding provides a scientific basis for further research into the therapeutic potential of luteolin in ameliorating myocardial injury.