IRF1 regulates autophagy and microglia polarization in retinal ischemia-reperfusion through NCOA1/Wnt/β-catenin signalling pathway

Abstract

Background

Interferon regulatory factor 1 (IRF1) is an important regulatory factor in the development of eyes, and it has been proved to be involved in the regulation of ischemia-reperfusion process. But its role in retinal ischemia-reperfusion (RIR) remains unclear.

Methods

RIR rat model was induced by increasing intraocular pressure. Hematoxylin and eosin (HE) staining, immunofluorescence (IF) staining, and western blot experiments were used to explore the levels of IRF1, autophagy, and microglia polarization in RIR. Western blot, transmission electron microscope, IF, and ELISA assays were used to explore the effects of IRF1, Nuclear receptor coactivator 1 (NCOA1), and Wnt/β-catenin signalling pathways in OGD/R-induced autophagy and polarization of rat retinal microglia. CHIP and dual-luciferase experiments verify the interaction between IRF1 and NCOA1. CHIP and dual-luciferase experiments were used to verify the interaction between IRF1 and NCOA1. Adeno-associated viruses interfering with IRF1 and NCOA1 were injected into the vitreous of rats to explore the functions of IRF1 and NCOA1 in RIR rats.

Results

IRF1 and M1-type markers of microglia in retina of RIR rats increased, and autophagy level decreased. Knockdown of IRF1 and NCOA1 increased autophagy of OGD/R-induced retinal microglia, inhibited M1-type polarization and inflammatory cytokines, alleviated RIR injury in rats, and inhibited the activation of Wnt/β-catenin signalling pathway. The Wnt/β-catenin signalling pathway activator HLY78 partially reversed the effect of knocking down NCOA1 on retinal microglia. Mechanically, knockdown of IRF1 inhibited the activation of Wnt/β-catenin signalling pathway by inhibiting the transcription of NCOA1.

Conclusion

Inhibition of IRF1 has a protective effect on RIR damage by regulating NCOA1/Wnt/β-catenin signalling pathway.