Anti-inflammatory properties of cytochrome P450 epoxygenase-derived eicosanoids

Science

Volumn 285, Issue 5431, 1999, Pages 1276-1279

  Node, Koichi ^a   Huo, Yuqing ^b   Ruan, Xiulu ^a   Yang, Baichun ^c   Spiecker, Martin ^a   Ley, Klaus ^b   Zeldin, Darryl C ^c   Liao, James K ^a  

^a BRIGHAM AND WOMEN S HOSPITAL   (United States)

^b UNIVERSITY OF VIRGINIA HEALTH SYSTEM   (United States)

^c NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CELL ADHESION MOLECULE; CYTOCHROME P450 ISOENZYME; EPOXIDE; EPOXYICOSATRIENOIC ACID; ICOSANOID; IMMUNOGLOBULIN ENHANCER BINDING PROTEIN; UNCLASSIFIED DRUG; VASCULAR CELL ADHESION MOLECULE 1;

ANTIINFLAMMATORY ACTIVITY; ARTICLE; DRUG MECHANISM; ENDOTHELIUM CELL; HUMAN; HUMAN CELL; IMMUNOHISTOCHEMISTRY; INFLAMMATION; LEUKOCYTE ADHERENCE INHIBITION; MOLECULAR CLONING; NUCLEOTIDE SEQUENCE; PRIORITY JOURNAL; VASODILATATION;

8,11,14-EICOSATRIENOIC ACID; ANIMALS; ANTI-INFLAMMATORY AGENTS, NON-STEROIDAL; CAROTID ARTERIES; CATTLE; CELL ADHESION; CELL ADHESION MOLECULES; CELLS, CULTURED; CORONARY VESSELS; CYTOCHROME P-450 ENZYME SYSTEM; DNA-BINDING PROTEINS; ENDOTHELIUM, VASCULAR; HUMANS; HYDROXYEICOSATETRAENOIC ACIDS; I-KAPPA B KINASE; I-KAPPA B PROTEINS; MICE; MICE, INBRED C57BL; NF-KAPPA B; OXYGENASES; PROTEIN-SERINE-THREONINE KINASES; TUMOR NECROSIS FACTOR-ALPHA; VASCULAR CELL ADHESION MOLECULE-1;

EID: 0033588034     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.285.5431.1276     Document Type: Article

References (26)

1. J. H. Capdevila, J. R. Falck, R. W. Estabrook, FASEB J. 6, 731 (1992).
2. W. B. Campbell and D. R. Harder, Circ. Res. 84, 484 (1999).
3. W. B. Campbell, D. Gebremedhin, P. F. Pratt, D. R. Harder, ibid. 78, 415 (1996).
4. M. Rosolowsky and W. B. Campbell, Biochim. Biophys. Acta 1299, 267 (1996).
5. C. L. Oltman, N. L. Weintraub, M. VanRollins, K. C. Dellsperger, Circ. Res. 83, 932 (1998).
6. S. L. Ptister, J. R. Falck, W. B. Campbell, Am. J. Physiol. 261, H843 (1991).
7. K. A. Pritchard Jr., P. V. Wong, M. B. Stemerman, Am. J. Pathol. 136, 1383 (1990).
8. S. Wu et al., J. Biol. Chem. 272, 12551 (1997).
9. S. Wu, C. R. Moomaw, K. B. Tomer, J. R. Falck, D. C Zeldin, ibid. 271, 3460 (1996).
10. Using 15 different pairs of degenerate oligonucleotides corresponding to conserved regions of cytochrome P450 epoxygenases, we performed RT-PCR on mRNA obtained from cultured human endothelial cells. Only one pair of primers (sense: 5′-GCTGACTTTCTCAAAAGACG-3′; antisense: 5′-CTCTGCACCTCATGGATGAC-3′; annealing/elongation temperature: 51°/72°C, 35 cycles) yielded a single 1-kb band that is identical to the gene encoding CYP2J2 (9).
11. Tissues were fixed in 10% neutral buffered formalin, processed, and embedded in paraffin. An affinity-purified rabbit polyclonal antiserum to CYP2J2 (anti-CYP2J2, 1:200 dilution) was used to detect CYP2J2 on serial sections (5 to 6 mm) of human heart (9). The specificity of anti-CYP2J2 was demonstrated by immunoblots of microsomal and S9 fractions prepared from various human tissues, in which anti-CYP2J2 produced a single band at 56 kD corresponding to endogenous CYP2J2 protein, and the finding that anti-CYP2J2 strongly bound recombinant CYP2J2 but did not recognize other cytochrome P450 isoforms including members of the CYP1A, CYP2A, CYP2B, CYP2C, CYP2D, and CYP2E subfamilies (8,9) and CYP4A1, CYP4A2, CYP4A3, CYP4A8, and CYP4A11.
12. F. M. Farin, T. H. Pohlman, C. J. Omiecinski, Toxicol. Appl. Pharmacol. 124, 1 (1994).
13. J. H.-C. Lin et al., Endothelium 4, 219 (1996).
14. B. G. Hoebel, E. Steyrer, W. F. Graier, Clin. Exp. Pharmacol. Physiol. 25, 826 (1998).
15. 2 flask) and unlabeled arachidonic acid (10 μM) in serum-free medium at 37°C. In some experiments, SKF-525A (100 μM) was added before the addition of arachidonic acid. At various time points, the media and cells were removed and extracted with diethyl ether. The combined organic phases were dried under a nitrogen stream, resolved by reversed-phase high-performance liquid chromatography (HPLC), and quantified by liquid scintillation as described (9). Products were identified by comparing their HPLC properties with those of authentic EET, DHET, HETE, and prostaglandin (PG) standards.
16. R. De Caterina et al., J. Clin. Invest. 96, 60 (1995).
17. M. VanRollins, T. L. Kaduce, H. R. Knapp, A. A. Spector, J. Lipid Res. 34, 1931 (1993).
18. V. J. Palombella, O. J. Rando, A. L. Goldberg, T. Maniatis, Cell 78, 773 (1994).
19. C. L Ramos et al., Circ. Res. 84, 1237 (1999).
20. 2. Cell rolling and adhesion were recorded on videotape by means of stroboscopic epifluoresence illumination with an intravital microscope (Zeiss ×20 objective, 0.5 numerical aperture). In mice receiving TNF-α only, the isolated vessels were also perfused with monoclonal antibody (mAb) MK-2 (40 μg/ml) for 20 min to block VCAM-1 function (21).
21. K. Miyake et al., J. Cell Biol. 114, 557 (1991).
22. T. Nakashima, Y. Harada, S. Miyata, T. Kiyohara, Am. J. Physiol. 271, R1274 (1996).
23. W. Kozak et al., ibid. 275, R1031 (1998).
24. J. K. Liao, unpublished observation.
25. Immunostaining for VCAM-1 expression was performed on paraffin sections (5 μm thick) of murine carotid arteries. Slides were incubated with avidinbiotin blocking reagent containing 10% horse serum (Vector Laboratories, Burlingame, CA) to reduce background staining, then incubated with primary antibody (polyctonal goat anti-mouse VCAM-1, 5 μg/ml; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4°C followed by biotin-conjugated horse anti-goat immunoglobulin C (IgC), ABC complex, and 3,3′-diaminobenzidíne as substrate (Vector Laboratories). All antibody incubations were performed in the presence of 10% horse serum. Slides were counterstained with hematoxytin and dehydrated in ethanol followed by xylene. Slides were examined under a light microscope (Zeiss × 100/1.4 oil immersion objective).
26. We thank J. R. Falck for EET and DHET standards and advice on EET stability in vivo; J. DiDonato and M. Karin for GST-IκB-α proteins; J. M. Sanders for immunostaining; J. Ma and J. Foley for technical assistance; and J. Bonner and J. Cidlowski for helpful suggestions. Supported by NIH Grant HL-52233 (J.K.L), HL-58108 (K.L), and NIEHS Division of Intramural Research (B.Y. and D.C.Z.), M.S. is a recipient of the Feodor Lynen Fellowship (Alexander von Humboldt-Stiftung). J.K.L is an Established Investigator, and K.N. is a Circulation Council/Otsuka Research Fellow of the American Heart Association.