Abstract
BACKGROUND: Uveitis is an autoimmune disease characterized by iris, ciliary muscle, and choroid inflammation. miR-223-3p, an anti-inflammatory microRNA, can regulate the expression of inflammatory genes involved in the disease process. However, the role and potential mechanism of miR-223-3p against uveitis remain unclear. METHODS: STRING website prediction, molecular docking, and co-IP experiments were performed to verify whether there was an interaction between RBPJ-HDAC1, HDAC1-P65, and P65-ARG1. Based on ex vivo and in vivo experiments, we detected NF-κB P65(lys310) acetylation, P65 nuclear translocation, and the level of M1/M2 macrophage polarization. In addition, we also determined the levels of mitochondrial pressure and calcium flux under different conditions. Regulation of NF-κB P65(lys310) acetylation via the RBPJ/HDAC1 axis affects macrophage polarization and mitochondrial function. RESULTS: We first found reduced miR-223-3p expression, elevated RBPJ and total acetylation levels, and an imbalance in macrophage polarization in peripheral blood monocyte-derived macrophages from patients with uveitis. Co-IP experiments supported the interaction between RBPJ and HDAC1, and HDAC1, as a key deacetylase, could inhibit NF-κB P65(lys310) acetylation. Notably, overactivation of NF-κB P65(lys310) acetylation levels in uveitis leads to elevated polarization of M1 macrophages and mitochondrial dysfunction. miR-223-3p can attenuate NF-κB P65(lys310) acetylation and P65 nuclear translocation levels through the RBPJ/HDAC1 axis in uveitis, effectively reducing pro-inflammatory macrophage levels and mitigating mitochondrial damage, thereby reducing ocular inflammation and positively regulating the intraocular microenvironment in uveitis. CONCLUSION: miR-223-3p can inhibit P65(lys310) acetylation and enhance mitochondrial function to improve M1/M2 macrophage polarization balance to ameliorate uveitis through RBPJ/HDAC1 axis. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s11658-026-00874-3.