Abstract
INTRODUCTION: The NLRP3 inflammasome/IL-1β-dependent inflammatory response serves as a critical factor and key trigger in exacerbating atherosclerosis (AS), whereas chaperone-mediated autophagy (CMA) recognizes and degrades the NLRP3 inflammasome. Targeting this pathway represents a more nuanced and targeted anti-inflammatory strategy to mitigate AS progression. As a key bioactive component derived from Ginkgo biloba leaves, Ginkgolide C (GC) possesses notable anti-inflammatory effects and confers protection against myocardial and cerebral ischemia-reperfusion injuries. The current research aimed to investigate whether GC could exert protective effects against AS and to elucidate its potential underlying mechanisms. METHOD: This study established both in vivo (high-fat diet/vitamin D3-induced atherosclerotic mouse model) and in vitro (LPS/ATP-stimulated RAW264.7 macrophage injury model) systems. In vivo evaluations included: H&E and Oil Red O staining for atherosclerotic lesion assessment; biochemical detection for lipid profiles; transmission electron microscopy for autophagic structure observation; immunohistochemistry and immunofluorescence for CMA regulator (LAMP-2A), NLRP3 inflammasome as well as key pro-inflammatory cytokines such as IL-1β, IL-18, and TNF-α. In vitro analyses comprised: MTT assay for cell viability; ELISA for quantifying inflammatory cytokine secretion; Western blotting for LAMP-2A, NLRP3 inflammasome, and NF-κB, MAPK signaling pathways molecules. LAMP-2A knockdown was conducted using siRNA to validate the CMA-dependent mechanisms underlying GC's effects. RESULT: Our results demonstrate that GC potentiated CMA activity in macrophages, leading to promoted degradation of the NLRP3 inflammasome via the lysosomal pathway. This process effectively suppressed the NLRP3 inflammasome/IL-1β-driven inflammatory cascade, ultimately attenuating atherosclerotic progression. CONCLUSION: GC alleviates AS via a novel LAMP-2A-dependent mechanism that enhances protein clearance and suppresses NLRP3 inflammation, providing a targeted alternative to broad immunosuppression. These results establish GC as a promising therapeutic candidate and prompt further studies on its clinical efficacy and applicability in other chronic inflammatory diseases.