Skip to main content
NCBI home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1990 Jun;136(6):1383–1391.

Atherogenic concentrations of low-density lipoprotein enhance endothelial cell generation of epoxyeicosatrienoic acid products.

K A Pritchard Jr 1, P Y Wong 1, M B Stemerman 1
PMCID: PMC1877592  PMID: 2356865

Abstract

To investigate the effects of protracted low-density lipoprotein (LDL) exposure on endothelial cell (EC) epoxyeicosatrienoic acid (EET) generation, human umbilical vein ECs were incubated in atherogenic concentrations of LDL (240 mg cholesterol per deciliter) (LDL-EC). After 4 days' incubation with LDL, EC were stimulated with human thrombin in the presence of 1-[14C]-arachidonic acid. Substantially more EET products were generated by LDL-ECs than by cells not exposed to high levels of LDL (C-EC). Thrombin stimulation caused LDL-EC to produce five- to eightfold more in 14,15-EET, 11,12-EET, 8,9-EET, and 5,6-EET, with 14,15-EET as the major product. This is the first demonstration, to date, that EETs can be induced in EC. Metapyrone (SKF-525A) markedly inhibited EC EET generation, indicating a role for the cytochrome P-450 enzyme system in human EC arachidonic acid metabolism. One EET product, 14,15-EET, has been found to be chemotactic and to promote adhesion of U937 cells, a human monocytic lymphoma cell line, to EC. Thus, protracted exposure to atherogenic LDL concentrations increases the generation of chemotactic and adhesion factors (ie, 14,15-EET) after thrombin stimulation, possibly through the cytochrome P-450 enzyme system.

Full text

PDF
1383

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abraham N. G., Pinto A., Levere R. D., Mullane K. Identification of heme oxygenase and cytochrome P-450 in the rabbit heart. J Mol Cell Cardiol. 1987 Jan;19(1):73–81. doi: 10.1016/s0022-2828(87)80546-1. [DOI] [PubMed] [Google Scholar]
  2. Abraham N. G., Pinto A., Mullane K. M., Levere R. D., Spokas E. Presence of cytochrome P-450-dependent monooxygenase in intimal cells of the hog aorta. Hypertension. 1985 Nov-Dec;7(6 Pt 1):899–904. doi: 10.1161/01.hyp.7.6.899. [DOI] [PubMed] [Google Scholar]
  3. Ballou L. R., Lam B. K., Wong P. Y., Cheung W. Y. Formation of cis-14,15-oxido-5,8,11-icosatrienoic acid from phosphatidylinositol in human platelets. Proc Natl Acad Sci U S A. 1987 Oct;84(20):6990–6994. doi: 10.1073/pnas.84.20.6990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Capdevila J., Chacos N., Werringloer J., Prough R. A., Estabrook R. W. Liver microsomal cytochrome P-450 and the oxidative metabolism of arachidonic acid. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5362–5366. doi: 10.1073/pnas.78.9.5362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cunningham F. M., Woollard P. M. 12(R)-hydroxy-5,8,10,14-eicosatetraenoic acid is a chemoattractant for human polymorphonuclear leucocytes in vitro. Prostaglandins. 1987 Jul;34(1):71–78. doi: 10.1016/0090-6980(87)90264-4. [DOI] [PubMed] [Google Scholar]
  6. DiCorleto P. E., Chisolm G. M., 3rd Participation of the endothelium in the development of the atherosclerotic plaque. Prog Lipid Res. 1986;25(1-4):365–374. doi: 10.1016/0163-7827(86)90074-3. [DOI] [PubMed] [Google Scholar]
  7. DiCorleto P. E., de la Motte C. A. Characterization of the adhesion of the human monocytic cell line U937 to cultured endothelial cells. J Clin Invest. 1985 Apr;75(4):1153–1161. doi: 10.1172/JCI111810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fleisher L. N., Tall A. R., Witte L. D., Miller R. W., Cannon P. J. Stimulation of arterial endothelial cell prostacyclin synthesis by high density lipoproteins. J Biol Chem. 1982 Jun 25;257(12):6653–6655. [PubMed] [Google Scholar]
  9. Friedewald W. T., Levy R. I., Fredrickson D. S. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem. 1972 Jun;18(6):499–502. [PubMed] [Google Scholar]
  10. Gerrity R. G., Goss J. A., Soby L. Control of monocyte recruitment by chemotactic factor(s) in lesion-prone areas of swine aorta. Arteriosclerosis. 1985 Jan-Feb;5(1):55–66. doi: 10.1161/01.atv.5.1.55. [DOI] [PubMed] [Google Scholar]
  11. Gerrity R. G. The role of the monocyte in atherogenesis: I. Transition of blood-borne monocytes into foam cells in fatty lesions. Am J Pathol. 1981 May;103(2):181–190. [PMC free article] [PubMed] [Google Scholar]
  12. Gorman R. R., Oglesby T. D., Bundy G. L., Hopkins N. K. Evidence for 15-HETE synthesis by human umbilical vein endothelial cells. Circulation. 1985 Oct;72(4):708–712. doi: 10.1161/01.cir.72.4.708. [DOI] [PubMed] [Google Scholar]
  13. Hajjar D. P., Marcus A. J., Hajjar K. A. Interactions of arterial cells. Studies on the mechanisms of endothelial cell modulation of cholesterol metabolism in co-cultured smooth muscle cells. J Biol Chem. 1987 May 25;262(15):6976–6981. [PubMed] [Google Scholar]
  14. Hajjar D. P., Weksler B. B. Metabolic activity of cholesteryl esters in aortic smooth muscle cells is altered by prostaglandins I2 and E2. J Lipid Res. 1983 Sep;24(9):1176–1185. [PubMed] [Google Scholar]
  15. Holland J. A., Pritchard K. A., Rogers N. J., Stemerman M. B. Perturbation of cultured human endothelial cells by atherogenic levels of low density lipoprotein. Am J Pathol. 1988 Sep;132(3):474–478. [PMC free article] [PubMed] [Google Scholar]
  16. Honn K. V., Grossi I. M., Diglio C. A., Wojtukiewicz M., Taylor J. D. Enhanced tumor cell adhesion to the subendothelial matrix resulting from 12(S)-HETE-induced endothelial cell retraction. FASEB J. 1989 Sep;3(11):2285–2293. doi: 10.1096/fasebj.3.11.2673900. [DOI] [PubMed] [Google Scholar]
  17. Johnson A. R., Revtyak G., Campbell W. B. Arachidonic acid metabolites and endothelial injury: studies with cultures of human endothelial cells. Fed Proc. 1985 Jan;44(1 Pt 1):19–24. [PubMed] [Google Scholar]
  18. Jolly S. R., Lucchesi B. R. Effect of BW755C in an occlusion-reperfusion model of ischemic myocardial injury. Am Heart J. 1983 Jul;106(1 Pt 1):8–13. doi: 10.1016/0002-8703(83)90431-3. [DOI] [PubMed] [Google Scholar]
  19. Joris I., Zand T., Nunnari J. J., Krolikowski F. J., Majno G. Studies on the pathogenesis of atherosclerosis. I. Adhesion and emigration of mononuclear cells in the aorta of hypercholesterolemic rats. Am J Pathol. 1983 Dec;113(3):341–358. [PMC free article] [PubMed] [Google Scholar]
  20. Kelley J. L., Rozek M. M., Suenram C. A., Schwartz C. J. Activation of human peripheral blood monocytes by lipoproteins. Am J Pathol. 1988 Feb;130(2):223–231. [PMC free article] [PubMed] [Google Scholar]
  21. Kutsky P., Falck J. R., Weiss G. B., Manna S., Chacos N., Capdevila J. Effects of newly reported arachidonic acid metabolites on microsomal Ca++ binding, uptake and release. Prostaglandins. 1983 Jul;26(1):13–21. doi: 10.1016/0090-6980(83)90070-9. [DOI] [PubMed] [Google Scholar]
  22. Laniado-Schwartzman M., Davis K. L., McGiff J. C., Levere R. D., Abraham N. G. Purification and characterization of cytochrome P-450-dependent arachidonic acid epoxygenase from human liver. J Biol Chem. 1988 Feb 15;263(5):2536–2542. [PubMed] [Google Scholar]
  23. Maciag T., Hoover G. A., Stemerman M. B., Weinstein R. Serial propagation of human endothelial cells in vitro. J Cell Biol. 1981 Nov;91(2 Pt 1):420–426. doi: 10.1083/jcb.91.2.420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morel D. W., DiCorleto P. E., Chisolm G. M. Endothelial and smooth muscle cells alter low density lipoprotein in vitro by free radical oxidation. Arteriosclerosis. 1984 Jul-Aug;4(4):357–364. doi: 10.1161/01.atv.4.4.357. [DOI] [PubMed] [Google Scholar]
  25. Morel D. W., Hessler J. R., Chisolm G. M. Low density lipoprotein cytotoxicity induced by free radical peroxidation of lipid. J Lipid Res. 1983 Aug;24(8):1070–1076. [PubMed] [Google Scholar]
  26. Mullane K. M., Moncada S. The salvage of ischaemic myocardium by BW755C in anaesthetised dogs. Prostaglandins. 1982 Aug;24(2):255–266. doi: 10.1016/0090-6980(82)90151-4. [DOI] [PubMed] [Google Scholar]
  27. Munro J. M., Cotran R. S. The pathogenesis of atherosclerosis: atherogenesis and inflammation. Lab Invest. 1988 Mar;58(3):249–261. [PubMed] [Google Scholar]
  28. Murphy R. C., Falck J. R., Lumin S., Yadagiri P., Zirrolli J. A., Balazy M., Masferrer J. L., Abraham N. G., Schwartzman M. L. 12(R)-hydroxyeicosatrienoic acid: a vasodilator cytochrome P-450-dependent arachidonate metabolite from the bovine corneal epithelium. J Biol Chem. 1988 Nov 15;263(32):17197–17202. [PubMed] [Google Scholar]
  29. Pomerantz K. B., Fleisher L. N., Tall A. R., Cannon P. J. Enrichment of endothelial cell arachidonate by lipid transfer from high density lipoproteins: relationship to prostaglandin I2 synthesis. J Lipid Res. 1985 Oct;26(10):1269–1276. [PubMed] [Google Scholar]
  30. Pomerantz K. B., Hajjar D. P. Eicosanoid metabolism in cholesterol-enriched arterial smooth muscle cells: reduced arachidonate release with concomitant decrease in cyclooxygenase products. J Lipid Res. 1989 Aug;30(8):1219–1231. [PubMed] [Google Scholar]
  31. Powell W. S. Rapid extraction of arachidonic acid metabolites from biological samples using octadecylsilyl silica. Methods Enzymol. 1982;86:467–477. doi: 10.1016/0076-6879(82)86218-6. [DOI] [PubMed] [Google Scholar]
  32. Pritchard K. A., Jr, Holland J. A., Rogers N. J., Crean C. C., Britton T. E., Onigman P., Stemerman M. B. Low-density lipoprotein preparation by combined diafiltration and ultracentrifugation. Anal Biochem. 1988 Oct;174(1):121–127. doi: 10.1016/0003-2697(88)90525-8. [DOI] [PubMed] [Google Scholar]
  33. Pritchard K. A., Jr, Tota R. R., Stemerman M. B., Wong P. Y. 14, 15-Epoxyeicosatrienoic acid promotes endothelial cell dependent adhesion of human monocytic tumor U937 cells. Biochem Biophys Res Commun. 1990 Feb 28;167(1):137–142. doi: 10.1016/0006-291x(90)91741-a. [DOI] [PubMed] [Google Scholar]
  34. Proctor K. G., Capdevila J. H., Falck J. R., Fitzpatrick F. A., Mullane K. M., McGriff J. C. Cardiovascular and renal actions of cytochrome P-450 metabolites of arachidonic acid. Blood Vessels. 1989;26(1):53–64. [PubMed] [Google Scholar]
  35. Revtyak G. E., Hughes M. J., Johnson A. R., Campbell W. B. Histamine stimulation of prostaglandin and HETE synthesis in human endothelial cells. Am J Physiol. 1988 Aug;255(2 Pt 1):C214–C225. doi: 10.1152/ajpcell.1988.255.2.C214. [DOI] [PubMed] [Google Scholar]
  36. Revtyak G. E., Johnson A. R., Campbell W. B. Cultured bovine coronary arterial endothelial cells synthesize HETEs and prostacyclin. Am J Physiol. 1988 Jan;254(1 Pt 1):C8–19. doi: 10.1152/ajpcell.1988.254.1.C8. [DOI] [PubMed] [Google Scholar]
  37. Rosenfeld M. E., Tsukada T., Gown A. M., Ross R. Fatty streak initiation in Watanabe Heritable Hyperlipemic and comparably hypercholesterolemic fat-fed rabbits. Arteriosclerosis. 1987 Jan-Feb;7(1):9–23. doi: 10.1161/01.atv.7.1.9. [DOI] [PubMed] [Google Scholar]
  38. Salari H., Braquet P., Borgeat P. Comparative effects of indomethacin, acetylenic acids, 15-HETE, nordihydroguaiaretic acid and BW755C on the metabolism of arachidonic acid in human leukocytes and platelets. Prostaglandins Leukot Med. 1984 Jan;13(1):53–60. doi: 10.1016/0262-1746(84)90102-1. [DOI] [PubMed] [Google Scholar]
  39. Siess M. H., Vernevaut M. F., Suschetet M. Influence du cholestérol et de la nature des acides gras alimentaires sur les activités de quelques monoxygénases microsomales du foie chez le rat. Reprod Nutr Dev. 1983;23(2A):195–206. [PubMed] [Google Scholar]
  40. Spector A. A., Scanu A. M., Kaduce T. L., Figard P. H., Fless G. M., Czervionke R. L. Effect of human plasma lipoproteins on prostacyclin production by cultured endothelial cells. J Lipid Res. 1985 Mar;26(3):288–297. [PubMed] [Google Scholar]
  41. Steinbrecher U. P., Parthasarathy S., Leake D. S., Witztum J. L., Steinberg D. Modification of low density lipoprotein by endothelial cells involves lipid peroxidation and degradation of low density lipoprotein phospholipids. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3883–3887. doi: 10.1073/pnas.81.12.3883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Steinbrecher U. P. Role of superoxide in endothelial-cell modification of low-density lipoproteins. Biochim Biophys Acta. 1988 Mar 4;959(1):20–30. doi: 10.1016/0005-2760(88)90145-2. [DOI] [PubMed] [Google Scholar]
  43. van Hinsbergh V. W., Scheffer M., Havekes L., Kempen H. J. Role of endothelial cells and their products in the modification of low-density lipoproteins. Biochim Biophys Acta. 1986 Aug 14;878(1):49–64. doi: 10.1016/0005-2760(86)90343-7. [DOI] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES