Transfection of Cytochrome P4504B1 into the Cornea Increases Angiogenic Activity of the Limbal Vessels

摘要: Injury to the ocular surface induces production of corneal epithelial-derived 12-hydroxyeicosatetrienoic acid (12-HETrE), which exhibits stereospecific potent inflammatory and angiogenic properties is formed by a cytochrome P450 (P450) enzyme, CYP4B1. We have cloned rabbit CYP4B1 into expression plasmid pIRES2-enhanced green fluorescent protein (EGFP) examined effect overexpression on inflammation in vivo limbal vessel sprouting ex vivo. Cultured epithelial cells transfected with pIRES2-EGFP-CYP4B1 metabolized arachidonic 12-HETrE at rate five times higher than that pIRES2-EGFP-transfected (3.53 ± 0.08 versus 0.62 0.10 nmol/h/106 cells; mean S.E.M., n = 6, p < 0.05), indicating functional Injection either cornea resulted EGFP fluorescence epithelium. However, neovascularization, as measured length penetrating blood vessels, was significantly greater corneas eyes pIRES2-CYP4B1 compared pIRES2-EGFP. Corneal-limbal explants from showed marked activity (46 10 12 3 mm capillary length, correlated increased levels 12-HETrE, CYP4B1-derived 12-hydroxyeicosanoid (0.93 0.18 0.15 0.02 pmol/explant, inhibited (76 5%) inhibitor 17-octadecynoic acid. The results further implicate component cascade initiated injury raise possibility new therapeutic target for preventing namely, CYP4B1-12-HETrE system.