Cell surface nucleolin promotes endothelial cell pyroptosis in atherosclerosis through RASSF2.

Abstract

BACKGROUND AND AIMS: Increasing evidence indicates that modulating pyroptosis in endothelial cells (ECs) can alleviate atherosclerosis (AS) progression; however, despite reports that nucleolin (NCL) regulates vascular smooth muscle cell proliferation in AS, the potential mechanism by which cell surface NCL mediates pyroptosis in ECs during AS remains poorly understood. METHODS: AS was induced in ApoEmice by feeding a high-fat diet, after which aortic lesions were evaluated. Pyroptosis, inflammatory status, and NCL expression in ECs of the aortic root were then assessed. The effects of NLRP3 inflammasome inhibition and NCL modulation on atherosclerotic lesion severity in AS mice, as well as on pyroptosis in ox-LDL-stimulated ECs, were systematically investigated. In addition, the mechanistic role of NCL in AS was further explored using approaches including immunoprecipitation-mass spectrometry (IP-MS). RESULTS: AS model mice developed severe aortic lesions accompanied by pronounced EC pyroptosis and inflammation, together with elevated NCL expression in ECs of the aortic root. Both inhibition of NLRP3 and NCL knockdown alleviated atherosclerotic lesion severity in ApoEmice and attenuated ox-LDL-induced EC pyroptosis. Mechanistically, cell-surface NCL interacted with RASSF2 via its RNA-binding domain, and suppression of NCL decreased nuclear RASSF2 expression. NCL facilitated the translocation of RASSF2 into the nucleus, thereby exacerbating EC pyroptosis and amplifying inflammatory responses. CONCLUSIONS: This study demonstrates that, in AS, NCL exacerbates EC pyroptosis and promotes disease progression by facilitating nuclear transport of RASSF2. This study defines the mechanistic roles of NCL in AS, thereby identifying a new molecular pathway and suggesting potential therapeutic targets.