Transformation of arachidonic acid by 5- and 15-lipoxygenase pathways in bovine adrenal fasciculata cells

Prostaglandins Leukot Essent Fatty Acids. 1990 Jun;40(2):93-102. doi: 10.1016/0952-3278(90)90150-j.

Abstract

[1-14C]Arachidonic acid was incubated with isolated bovine adrenal fasciculata cells for 15 min at 37gC. The metabolites were separated and purified by reverse- and straight-phase high performance liquid chromatography, and identified by gas chromatography-mass spectrometry or radioimmunoassay. Identified metabolites were 5-hydroxy-6,8,11,14-eicosatetraenoic acid (5-HETE), 15-hydroxy-5,8,11,13-eicosatetraenoic acid (15-HETE), leukotriene B4 and 11,14,15-trihydroxy-5,8,12-eicosatrienoic acid (11,14,15-THET). Addition of 15-hydroperoxy-5,8,11,13-eicosatetraenoic acid (15-HPETE), an intermediate metabolite of 15-lipoxygenase pathway to microsomes of bovine adrenal fasciculata cells resulted in the formation of 11,14,15-THET. The formation of 11,14,15-THET by microsomes was not dependent on the presence of NADPH, while it was dose-dependently suppressed by ketoconazole, a potent inhibitor of cytochrome P-450 dependent enzymes. These results indicate that 5- and 15-lipoxygenase pathways of arachidonic acid may exist in bovine adrenal fasciculata cells and that 15-HPETE is further metabolized to 11,14,15-THET by adrenal microsomal cytochrome P-450.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arachidonate 15-Lipoxygenase / metabolism*
  • Arachidonate 5-Lipoxygenase / metabolism*
  • Arachidonate Lipoxygenases / metabolism*
  • Arachidonic Acids / metabolism*
  • Cattle
  • Chemical Phenomena
  • Chemistry
  • Chromatography, High Pressure Liquid
  • Cytochrome P-450 Enzyme Inhibitors
  • In Vitro Techniques
  • Ketoconazole / pharmacology
  • Male
  • Microsomes / drug effects
  • Microsomes / enzymology
  • Rats
  • Rats, Inbred Strains
  • Zona Fasciculata / cytology
  • Zona Fasciculata / drug effects
  • Zona Fasciculata / enzymology*

Substances

  • Arachidonic Acids
  • Cytochrome P-450 Enzyme Inhibitors
  • Arachidonate Lipoxygenases
  • Arachidonate 15-Lipoxygenase
  • Arachidonate 5-Lipoxygenase
  • Ketoconazole