Acute glaucoma’s main sign is acute ocular hypertension (AOH), leading to retinal ganglion cell (RGC) death and irreversible visual loss. However, there is currently no approved effective therapy for this condition. This research aimed to identify the major regulators and the overall protein changes involved in AOH-induced RGC death. Mass-spectrometry was used to analyze proteomic patterns in the retinal protein extracts from the AOH and sham-group, and then Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway studies were performed. In proteomics analysis, we identified 92 proteins in the AOH group compared to the control group, with 58 proteins being up-regulated and 34 proteins being down-regulated. Western blot and biochemical assay analyses identified changes in Fatty acid-binding protein 7 (FABP7), and caveolin-1(Cav-1) that were related to fatty acid metabolism and ocular inflammatory signaling. Moreover, variations in the expression of the proteins Galectin-1 (Gal-1), S100 calcium-binding protein A6 (S100a6), and Visinin-like protein-1 (VILIP) was shown, all of which were associated to the process of neuronal ischemia. Our investigation demonstrated that neuroinflammation and fatty acid metabolism were involved in retinal impairment following AOH, offering a potential therapeutic strategy for acute glaucoma.