Pseudane V alleviates ox-LDL-induced macrophage M1 polarization by inhibiting m6A modification of PPARGC1A

Atherosclerosis is driven by oxidized low-density lipoprotein (ox-LDL)-triggered macrophage malfunction, yet the precise pathways and effective counter-measures remain elusive. Here we delineated how N6-methyladenosine (m⁶A) RNA methylation governs ox-LDL-induced M1 macrophage polarization and evaluated Marine natural products for therapeutic intervention. Human monocytic THP-1 cells differentiated into M0 macrophages were treated with ox-LDL. M1/M2 polarization states were analyzed using flow cytometry, and changes in polarization markers were examined using quantitative real-time polymerase chain reaction (qRT-PCR). Global m6A changes were detected using m⁶A dot blot and quantification analysis. Methylated RNA immunoprecipitation Sequencing (MeRIP-seq) and RNA Sequencing (RNA-seq) were performed to identify downstream target genes of the ox-LDL-m⁶A pathway. The role of Peroxisome Proliferator-activated Receptor gamma coactivator 1-alpha (PPARGC1A) in ox-LDL-induced M1 polarization was analyzed. A high-throughput marine natural product library was employed to discover agents capable of reversing ox-LDL-driven macrophage polarization. ox-LDL induces macrophage M1 polarization, and M1 macrophages damage endothelial cells. ox-LDL-induced polarization of macrophages into the M1 phenotype was associated with increased global RNA m⁶A modification. High levels of m⁶A modification induced by ox-LDL suppressed PPARGC1A expression in a YTH domain family 2 (YTHDF2)-dependent manner, leading to M1 polarization. Pseudane V, a marine natural product, effectively reduced ox-LDL-induced M1 polarization and protected vascular endothelial cells by correcting abnormal m⁶A modification of PPARGC1A. In conclusion, ox-LDL induces m⁶A modification of PPARGC1A, suppressing its expression and promoting M1 polarization of macrophages, contributing to atherosclerosis development. Pseudane V counteracts excessive m⁶A modification caused by ox-LDL, preventing M1 polarization of macrophages. These findings suggest the potential use of Pseudane V in preventing atherosclerosis.