Proliferative vitreoretinopathy (PVR) was induced in a rat model using intravitreal dispase to study fibrosis- and inflammation-related proteome changes and to evaluate the therapeutic effect of topical 3’,4’-dihydroxyflavonol (DiOHF) eye drops. Retina and vitreous were collected 28 days post-injury from vehicle-, DiOHF- and methotrexate-treated eyes, alongside contralateral controls. In parallel, ARPE-19 cells were stimulated with TGFβ1 + PDGF-BB to model wound-healing responses in vitro, with or without DiOHF. Label-free DIA LC-MS/MS was performed (Orbitrap Exploris 480 for rat tissues; Orbitrap Eclipse for ARPE-19), and data were analyzed with Spectronaut directDIA against UniProt Rattus norvegicus or Homo sapiens canonical databases. The dataset contains the original instrument raw files and processed Spectronaut results for both the rat retina/vitreous and ARPE-19 experiments.