Corneal diffusion and metabolism of 12(R)-hydroxyeicosatetraenoic acid (12(R)HETE)

Purpose. To quantify the corneal diffusion and metabolism of tritiated 12(R)hydroxyeicosatetraenoic acid (12(R)HETE) in the in vitro-mounted rabbit cornea to determine if this compound or one of its metabolites can diffuse across the stroma to the corneal endothelium. Methods. The studies were performed in a Lucite block perfusion chamber by placing tritiated 12(R)HETE on the tear side of the cornea under the following conditions: (A) cornea completely intact (endothelium and epithelium present); (B) cornea with epithelium removed; and (C) cornea with both epithelium and endothelium removed. Radioactivity of 12(R)HETE and metabolites were measured in the different corneal layers and in the corneal perfusates using scintillation spectroscopy. 12(R)-hydroxyeicosatetraenoic and its metabolites were then quantified in the tissue perfusates using high performance liquid chromatography (HPLC) analysis. Results. 12(R)HETE is rapidly taken up and metabolized by the intact cornea to a number of more polar compounds including a metabolite which has spectral and retention time characteristics of 8(R)-hydroxyhexadecatrienoic acid (8(R)HHDTrE). Both 12(R)HETE and 8(R)HHDTrE can diffuse through the stroma to the endothelium. Corneas having the epithelium removed also allow diffusion of 12(R)HETE across the stroma; however, there is significantly less metabolism. When both the epithelium and endothelium are removed, 12(R)HETE is capable of diffusing across the stroma; however, there is Little metabolism, which suggests that the majority of 12(R)HETE metabolism occurs in the epithelium and, to a lesser degree, in the endothelium and stroma. Conclusions. 12(R)HETE and its metabolites are capable of diffusing from the epithelium through the cornea where they may adversely affect the endothelium.