CYP2J Subfamily Cytochrome P450s in the Gastrointestinal Tract: Expression, Localization, and Potential Functional Significance
- Darryl C. Zeldin1,
- Julie Foley2,
- Susan M. Goldsworthy4,
- Molly E. Cook1,
- James E. Boyle1,
- Jixiang Ma1,
- Cindy R. Moomaw1,
- Kenneth B. Tomer3,
- Charles Steenbergen5 and
- Shu Wu1
- Laboratories of 1Pulmonary Pathobiology (D.C.Z., M.E.C., J.E.B., J.M., C.R.M., S.W.), 2Experimental Pathology (J.F.), and 3Molecular Biophysics (K.B.T.), National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, 4Pathology Associates International (S.M.G.), Durham, North Carolina 27713, and5Department of Pathology (C.S.), Duke University Medical Center, Durham, North Carolina 27710
Abstract
Our laboratory recently described a new human cytochrome P450 arachidonic acid epoxygenase (CYP2J2) and the corresponding rat homologue (CYP2J3), both of which were expressed in extrahepatic tissues. Northern analysis of RNA prepared from the human and rat intestine demonstrated that CYP2J2 and CYP2J3 mRNAs were expressed primarily in the small intestine and colon. In contrast, immunoblotting studies using a polyclonal antibody raised against recombinant CYP2J2 showed that CYP2J proteins were expressed throughout the gastrointestinal tract. Immunohistochemical staining of formalin-fixed, paraffin-embedded intestinal sections using anti-CYP2J2 IgG and avidin-biotin-peroxidase detection revealed that CYP2J proteins were present at high levels in nerve cells of autonomic ganglia, epithelial cells, intestinal smooth muscle cells, and vascular endothelium. The distribution of this immunoreactivity was confirmed by in situ hybridization using a CYP2J2-specific antisense RNA probe. Microsomal fractions prepared from human jejunum catalyzed the NADPH-dependent metabolism of arachidonic acid to epoxyeicosatrienoic acids as the principal reaction products. Direct evidence for thein vivo epoxidation of arachidonic acid by intestinal cytochrome P450 was provided by documenting, for the first time, the presence of epoxyeicosatrienoic acids in human jejunum by gas chromatography/mass spectrometry. We conclude that human and rat intestine contain an arachidonic acid epoxygenase belonging to the CYP2J subfamily that is localized to autonomic ganglion cells, epithelial cells, smooth muscle cells, and vascular endothelium. In addition to the known effects on intestinal vascular tone, we speculate that CYP2J products may be involved in the release of intestinal neuropeptides, control of intestinal motility, and/or modulation of intestinal fluid/electrolyte transport.
Footnotes
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Send reprint requests to: Darryl C. Zeldin, M.D., NIH/NIEHS, 111 T.W. Alexander Drive, Bldg. 101, D236, Research Triangle Park, NC 27709. E-mail: zeldin{at}niehs.nih.gov
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↵1 The P450 nomenclature detailed in Ref. 40 is used throughout this report.
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↵2 J. Ma and D. C. Zeldin. Cloning, expression, and function of mouse CYP2J5 and CYP2J6. Manuscript in preparation.
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↵3 L. S. Kaminsky, personal communication.
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↵4 J. Ma and D. C. Zeldin, unpublished observations.
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This work was partially supported by National Institutes of Health Contract N01-ES35356 and the Bryan Alzheimer Disease Research Center Rapid Autopsy Program (NIH P50-AG05128).
- Abbreviations:
- P450
- cytochrome P450
- EET
- cis-epoxyeicosatrienoic acid
- DHET
- vic-dihydroxyeicosatrienoic acid
- AA
- arachidonic acid
- 20-OH-AA
- 20-hydroxyeicosatetraenoic acid
- HPLC
- high performance liquid chromatography
- GC/MS
- gas chromatography/mass spectrometry
- PFB
- pentafluorobenzyl
- SDS
- sodium dodecyl sulfate
- SSC
- standard saline citrate
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- Received September 20, 1996.
- Accepted February 14, 1997.
- The American Society for Pharmacology and Experimental Therapeutics
