Peli1, regulated by m 6 A modification, suppresses atherosclerosis progression by inhibiting YB-1-mediated NLRP3 inflammasome activation

Abstract

AbstractBackground The activation of NLRP3 inflammasome in macrophages is a risking factor accelerating atherosclerosis (AS) progression. Here, the function of Peli1 in regulating NLRP3 inflammasome activation during AS progression were investigated. MethodsApoE−/−mice were subjected to high fat diet to construct AS modelin vivo. HE, Oil red O and Sirius red staining were adopted to analyze histopathological changes and lipid accumulation. Raw264.7 cells stimulated by ox-LDL were used as AS cellular model. Pro-inflammatory cytokines secretion was assessed using ELISA. Total m6A level was examined by m6A dot blot assay, and Peli1 m6A level was assessed using MeRIP assay. The interactions between METTL3, YTHDC2, Peli1, YB-1 and NLRP3 were analyzed by ChIP, dual-luciferase reporter gene, CoIP, RIP and/or RNA pull down assays. Results YB-1 knockdown could inhibit AS progressionin vivo, and YB-1 silencing repressed ox-LDL-mediated lipid accumulation and inflammation in macrophages by inactivating NLRP3 inflammasome. E3 ubiquitination ligase Peli1 mediated ubiquitination degradation of YB-1 during AS progression. Moreover, it was found that YTHDC2 recognized METTL3-mediated Peli1 m6A modification and mediated Peli1 mRNA degradation. Rescue studies revealed that YB-1 upregulation abrogated Peli1 upregulation’s repression on AS progression bothin vitroandin vivo. Conclusion Peli1, regulated by m6A modification, inhibited YB-1-mediated NLRP3 inflammasome activation in macrophages by promoting YB-1 ubiquitination to suppress AS progression.