Neochlorogenic Acid Attenuates Endothelial Injury in Diabetic ApoE -/-Mice by Suppressing NF-κB Mediated NLRP3 Inflammasome Activation

Abstract

Diabetes mellitus (DM) significantly increases global morbidity and mortality through cardiovascular complications, particularly atherosclerosis, characterized by chronic inflammation, oxidative stress, and endothelial dysfunction. Recent studies suggest that pyroptosis, a proinflammatory form of programmed cell death regulated by the nuclear factor kappa-light-chainenhancer of activated B cells (NF-κB)/NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway, as a crucial contributor to diabetes-related vascular injury. However, therapeutic strategies that directly target pyroptosis remain limited. Neochlorogenic acid (nCGA), a naturally occurring polyphenolic compound abundant in mulberry leaves, coffee, and green tea, demonstrates potent antioxidant and anti-inflammatory activities, suggesting its therapeutic potential against diabetesassociated complications. In this study, bioinformatics analyses, molecular simulations, and experimental models were integrated to investigate the protective mechanisms of nCGA. The bioinformatics workflow included KEGG pathway enrichment, Gene Ontology annotation, and construction of protein-protein interaction networks, supported by molecular docking and molecular dynamics simulations to evaluate interactions between nCGA and potential regulatory targets. To verify these findings, in vitro experiments using glucolipotoxic endothelial cells and in vivo studies in diabetic ApoE -/-mice were performed. NF-κB was identified as a central therapeutic target of nCGA, and molecular docking revealed a stable and favorable binding interaction. In vitro, nCGA significantly attenuated glucolipotoxicity-induced oxidative stress, lipid accumulation, mitochondrial dysfunction, apoptosis, and pyroptosis by suppressing NF-κB/NLRP3 inflammasome activation. Consistently, in diabetic ApoE -/-mice, nCGA administration improved metabolic parameters, reduced vascular lipid deposition, and lowered serum low-density lipoprotein cholesterol (LDL-C) levels, accompanied by decreased expression of inflammatory markers. In conclusion, these findings demonstrate that nCGA is a promising natural therapeutic candidate for alleviating diabetes-associated vascular injury through targeted inhibition of NF-κB and the NLRP3 inflammasome signaling pathway.