GPR124 promotes NLRP3 inflammasome activation through impaired mitochondrial autophagy homeostasis in ox-LDL-treated endothelial cells

G protein-coupled receptor 124 (GPR124) has been implicated in endothelial dysfunction, but its role in ox-LDL-induced endothelial inflammatory injury remains incompletely understood. This study investigated whether GPR124 contributes to endothelial cell injury through disruption of mitochondrial autophagy homeostasis and subsequent activation of the NLRP3 inflammasome. Using an ox-LDL-treated EA.hy926 endothelial cell model, we found that GPR124 expression was significantly upregulated under injurious conditions. GPR124 overexpression aggravated ox-LDL-induced cellular dysfunction, as reflected by reduced proliferative activity, increased reactive oxygen species (ROS) production, and enhanced lipid accumulation. Mechanistically, GPR124 dysregulation was associated with impaired mitochondrial homeostasis, including loss of mitochondrial membrane potential, excessive ROS generation, and altered mitochondrial autophagy flux, accompanied by increased NLRP3 inflammasome activation and IL-1β release. In contrast, GPR124 knockdown partially attenuated these abnormalities and alleviated endothelial cell injury. In addition, pharmacological intervention with Mdivi-1 supported the involvement of mitochondrial autophagy-related processes in the inflammatory phenotype observed under ox-LDL stimulation. Bioinformatics analyses further suggested that GPR124-associated differentially expressed genes were enriched in pathways related to mitochondrial homeostasis, metabolic regulation, and inflammatory signaling. Taken together, these findings suggest that GPR124 may contribute to ox-LDL-induced endothelial inflammatory injury through dysregulation of the mitochondrial autophagy-NLRP3 axis and identify GPR124 as a potential target for further mechanistic investigation in endothelial dysfunction.