Proteomic Investigation of the Anti-Atherosclerotic Mechanism of 4',7-di- O-Methylnaringenin from Zingiberaceae Plants.

INTRODUCTION: Atherosclerosis (AS) is a chronic inflammatory disease driven by disordered lipid metabolism and is a major cause of cardiovascular disease. 4',7-Di-Omethylnaringenin (DMNG), a natural flavonoid derived from Zingiberaceae plants, has diverse biological activities, particularly anti-inflammatory and antioxidant effects. This study aimed to investigate the therapeutic effects of DMNG on AS and elucidate its underlying molecular mechanisms using proteomics. METHODS: C57BL/6 mice fed a standard diet served as normal controls, whereas ApoE-/- mice fed a high-fat diet (HFD) were used as an AS model. ApoE-/- mice were treated with DMNG (25, 50, or 100 mg/kg/day) for 12 weeks. Body weight, serum lipid profiles, inflammatory cytokines, and oxidative stress markers were measured. Histopathological changes in the aorta were assessed using H&E and Masson's trichrome staining. Proteomic analysis was performed to identify key proteins and pathways, followed by validation using Western blotting. RESULTS: DMNG treatment significantly reduced body weight gain and alleviated aortic intimal hyperplasia, plaque formation, and inflammatory infiltration while increasing collagen deposition. It improved lipid profiles by decreasing serum TC, TG, and LDL-C levels and increasing HDL-C levels. DMNG also reduced the serum levels of pro-inflammatory cytokines (IL-1β, IL- 6, and TNF-α), enhanced antioxidant enzyme activities (SOD and GSH-Px), and decreased MDA levels. Proteomic analysis identified ICAM1 as a protein downregulated by DMNG and implicated the MAPK/NF-κB pathway as the main regulatory axis. Western blotting confirmed that DMNG reduced the levels of ICAM1, p-p65, p-IκB, p-ERK, and p-p38. DISCUSSION: These findings identify ICAM1 as a critical factor through which DMNG exerts its anti-atherosclerotic effects. By suppressing ICAM1 expression and the MAPK/NF-κB signaling pathway, DMNG alleviates vascular inflammation and oxidative stress, two core pathological processes in AS. Given the incomplete efficacy and adverse effects of current lipid-lowering therapies in some patients, DMNG, a natural flavonoid with multi-target activity, may represent a therapeutic candidate for managing AS, particularly its inflammatory components. CONCLUSION: DMNG ameliorates atherosclerosis by suppressing ICAM1 expression and MAPK/NF-κB signaling.