Hypoxia stimulates the synthesis of cytochrome P450-derived inflammatory eicosanoids in rabbit corneal epithelium

C. Vafeas, P. A. Mieyal, F. Urbano, J R Falck, K. Chauhan, M. Berman, M. L. Schwartzman

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The corneal epithelium metabolizes arachidonic acid by a cytochrome P450-(CYP) mediated pathway to 12(R)hydroxy-5,8,10,14-eicosatrienoic acid [12(R)-HETE] and 12(R)hydroxy-5,8,14-eicosatrienoic acid [12(R)-HETrE]. Both metabolites possess potent inflammatory properties with 12(R)-HETrE being a powerful angiogenic factor and assume the role of inflammatory mediators in hypoxia- and chemical-induced injury in the cornea, in vivo. We developed an in vitro model of corneal organ culture to characterize the biochemical and molecular events involved in the increased synthesis of these metabolites. These cultured corneas exhibit epithelial cytochrome P450 CYP-dependent 12(R)-HETE and 12(R)-HETrE synthesis as indicated by chiral analysis and by the ability of CYP enzyme inhibitors to repress their synthesis. Hypoxia greatly and selectively stimulated the synthesis of 12(R)-HETE (7-fold over control normoxic conditions) and 12(R)-HETrE. The bacterial endotoxin, lipopolysaccharide, also increased the synthesis of these eicosanoids, substantiating the notion that this activity may function as an inflammatory pathway. These metabolites were detected in the culture medium by gas chromatography/mass spectroscopy (GC/MS) analysis and their levels significantly increased in hypoxia-treated corneas, further indicating their endogenous formation in response to injury. This in vitro model provides an excellent preparation for studying factors regulating the synthesis of these inflammatory eicosanoids and for isolating, identifying and characterizing the CYP protein responsible for their synthesis.

Original languageEnglish (US)
Pages (from-to)903-910
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume287
Issue number3
StatePublished - 1998

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Corneal Epithelium
Eicosanoids
Cytochrome P-450 Enzyme System
Rabbits
12-Hydroxy-5,8,10,14-eicosatetraenoic Acid
Cornea
Angiogenesis Inducing Agents
Organ Culture Techniques
Enzyme Inhibitors
Hypoxia
12-hydroxy-5,8,14-eicosatrienoic acid
Arachidonic Acid
Endotoxins
Gas Chromatography
Culture Media
Lipopolysaccharides
Mass Spectrometry
Wounds and Injuries

ASJC Scopus subject areas

  • Pharmacology

Cite this

Hypoxia stimulates the synthesis of cytochrome P450-derived inflammatory eicosanoids in rabbit corneal epithelium. / Vafeas, C.; Mieyal, P. A.; Urbano, F.; Falck, J R; Chauhan, K.; Berman, M.; Schwartzman, M. L.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 287, No. 3, 1998, p. 903-910.

Research output: Contribution to journalArticle

Vafeas, C. ; Mieyal, P. A. ; Urbano, F. ; Falck, J R ; Chauhan, K. ; Berman, M. ; Schwartzman, M. L. / Hypoxia stimulates the synthesis of cytochrome P450-derived inflammatory eicosanoids in rabbit corneal epithelium. In: Journal of Pharmacology and Experimental Therapeutics. 1998 ; Vol. 287, No. 3. pp. 903-910.
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