AIM: To observe the effects of the inhibition of NADPH oxidase 4 (NOX4) expression on the retinal vascular barriers and visual function after retinal detachment (RD). METHODS: RD model was established 3wk after adeno-associaned virus vector injection. The retinal tissue was harvested 3d after RD, and the death of retinal vascular endothelial cells and photoreceptors was observed using electron microscopy. The NOX4 expression was detected by Western blot. Confocal microscopy was used to observe a retinal patch that had been perfused with Evans blue. A modified water maze test was used to detect the time required to find the platform on the water surface. The visual function of the rats was evaluated and reactive oxygen species (ROS) expression was detected by a fluorescence microplate reader. RESULTS: The retinal patch showed that NOX4 interference significantly reduced the destruction of the tight junctions between the retinal endothelium of RD rats and reduced leakage. Western blotting showed decreased expression of the NOX4 protein and decreased expression of ROS in retinal tissue; the Morris water maze test results showed that NOX4 interference significantly decreased the escape latency of the rats. CONCLUSION: NOX4 interference reduces the production of ROS in retinal vascular endothelial cells after experimental RD, thereby protecting the blood-retinal barrier and protecting visual function.