Identification and localization of five CYP2Cs in murine extrahepatic tissues and their metabolism of arachidonic acid to regio- and stereoselective products

J Pharmacol Exp Ther. 2001 Oct;299(1):39-47.

Abstract

The CYP2C subfamily has been extensively studied in humans with respect to the metabolism of clinically important drugs, and polymorphisms have been identified in these enzymes. In the present study, a murine model was used to determine the possible physiological functions and extrahepatic distribution of CYP2Cs. Using the reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and immununohistochemistry, this report demonstrates that the mouse CYP2Cs are extensively distributed in extrahepatic tissues and localized to heart muscle, lung Clara and ciliated cells, kidney collecting ducts, the X-zone of female adrenals, reproductive organs, white blood cells, and eyes (in the optic nerve, rods, and cones). RT-PCR, subcloning, and sequencing of the products indicate that each CYP2C has a unique tissue distribution. Four cDNA fragments representing potentially new CYP2Cs were identified, each with its own organ-specific pattern of expression. Using a bacterial cDNA expression system, we found that recombinant proteins for each of the five full-length murine CYP2Cs metabolize arachidonic acid to different regio- and stereospecific products, including epoxyeicosatrienoic acids and hydroxyeicosatetraenoic acids. Regio- and stereospecific metabolites of arachidonic acid have been reported to affect important physiological functions such as inflammation, neutrophil activation, ion transport, cellular proliferation, and vascular tone. Our results suggest that the presence of CYP2C enzymes in heart muscle, aorta, kidney, lung, adrenals, eyes, and reproductive organs could regulate important physiological and/or pathological processes in these tissues.

MeSH terms

  • Animals
  • Arachidonic Acid / metabolism*
  • Blotting, Western
  • Cytochrome P-450 Enzyme System / metabolism*
  • Female
  • Hydroxyeicosatetraenoic Acids / metabolism
  • Immunohistochemistry
  • Isoenzymes / metabolism
  • Male
  • Mice
  • Microsomes / enzymology
  • RNA, Messenger / biosynthesis
  • Recombinant Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stereoisomerism

Substances

  • Hydroxyeicosatetraenoic Acids
  • Isoenzymes
  • RNA, Messenger
  • Recombinant Proteins
  • cytochrome P-450 CYP2C subfamily
  • Arachidonic Acid
  • Cytochrome P-450 Enzyme System