Abstract
The acute effect of in vitro deendothelialization on the production of prostacyclin (PGI2) by the rabbit aorta has been investigated. The effectiveness of removing endothelium by rubbing it against filter paper or scraping it with a scalpel was demonstrated by scanning electron microscopy and en face examination after silver staining. Endothelium removal produced an immediate stimulation of PGI2 release, resulting in 408% of the control after rubbing and 367% of the control after scraping, during the first 30-min period of incubation. This increased production of PGI2 gradually declined over time to reach values similar to the control after 2h. At that time, the deendothelialized aorta was totally unresponsive to the stimuli that increase PGI2 release in the intact aorta (acetylcholine, ADP, ionophore A23187, and arachidonic acid). The enhanced production of PGI2 in the deendothelialized aorta was associated with an increased release of free arachidonic acid (353% of the control): in contrast with PGI2, this stimulation was maintained for at least 150 min. A transient exposure of the deendothelialized aorta to ibuprofen (250 microM) was followed by a rebound of PGI2 production, which was also prolonged by BW-755C (3-10 microM). In conclusion, removal of the endothelium triggered an immediate and sustained mobilization of free arachidonic acid in the rabbit aorta: the resulting increase of PGI2 production was short-lived, probably as a consequence of cyclooxygenase self-inactivation. Our results indicate that the subendothelium has a significant capacity to produce PGI2, but that this capacity is expressed only briefly.
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- Beetens J. R., Claeys M., Herman A. G. Antioxidants increase the formation of 6-oxo-PGF1 alpha by ram seminal vesicle microsomes. Biochem Pharmacol. 1981 Oct;30(20):2811–2815. doi: 10.1016/0006-2952(81)90419-6. [DOI] [PubMed] [Google Scholar]
- Boeynaems J. M., Galand N. Cholinergic stimulation of vascular prostacyclin synthesis. Prostaglandins. 1983 Oct;26(4):531–544. doi: 10.1016/0090-6980(83)90191-0. [DOI] [PubMed] [Google Scholar]
- Boeynaems J. M., Galand N. Stimulation of vascular prostacyclin synthesis by extracellular ADP and ATP. Biochem Biophys Res Commun. 1983 Apr 15;112(1):290–296. doi: 10.1016/0006-291x(83)91829-6. [DOI] [PubMed] [Google Scholar]
- Brotherton A. F., Hoak J. C. Prostacyclin biosynthesis in cultured vascular endothelium is limited by deactivation of cyclooxygenase. J Clin Invest. 1983 Oct;72(4):1255–1261. doi: 10.1172/JCI111081. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Castellot J. J., Jr, Addonizio M. L., Rosenberg R., Karnovsky M. J. Cultured endothelial cells produce a heparinlike inhibitor of smooth muscle cell growth. J Cell Biol. 1981 Aug;90(2):372–379. doi: 10.1083/jcb.90.2.372. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chamley J. H., Campbell G. R., McConnell J. D., Gröschel-Stewart U. Comparison of vascular smooth muscle cells from adult human, monkey and rabbit in primary culture and in subculture. Cell Tissue Res. 1977 Feb 14;177(4):503–522. doi: 10.1007/BF00220611. [DOI] [PubMed] [Google Scholar]
- Chand N., Altura B. M. Acetylcholine and bradykinin relax intrapulmonary arteries by acting on endothelial cells: role in lung vascular diseases. Science. 1981 Sep 18;213(4514):1376–1379. doi: 10.1126/science.7268440. [DOI] [PubMed] [Google Scholar]
- Coughlin S. R., Moskowitz M. A., Antoniades H. N., Levine L. Serotonin receptor-mediated stimulation of bovine smooth muscle cell prostacyclin synthesis and its modulation by platelet-derived growth factor. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7134–7138. doi: 10.1073/pnas.78.11.7134. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Mey J. G., Claeys M., Vanhoutte P. M. Endothelium-dependent inhibitory effects of acetylcholine, adenosine triphosphate, thrombin and arachidonic acid in the canine femoral artery. J Pharmacol Exp Ther. 1982 Jul;222(1):166–173. [PubMed] [Google Scholar]
- DeWitt D. L., Day J. S., Sonnenburg W. K., Smith W. L. Concentrations of prostaglandin endoperoxide synthase and prostaglandin I2 synthase in the endothelium and smooth muscle of bovine aorta. J Clin Invest. 1983 Dec;72(6):1882–1888. doi: 10.1172/JCI111151. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dejana E., Balconi G., De Castellarnau C., Barbieri B., Vergara-Dauden M., De Gaetano G. Prostacyclin production by human endothelial and bovine smooth muscle cells in culture. Effect of repeated stimulation with arachidonic acid, thrombin and ionophore A23187. Biochim Biophys Acta. 1983 Feb 7;750(2):261–267. doi: 10.1016/0005-2760(83)90027-9. [DOI] [PubMed] [Google Scholar]
- Eldor A., Falcone D. J., Hajjar D. P., Minick C. R., Weksler B. B. Recovery of prostacyclin production by de-endothelialized rabbit aorta. Critical role of neointimal smooth muscle cells. J Clin Invest. 1981 Mar;67(3):735–741. doi: 10.1172/JCI110090. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FitzGerald G. A., Smith B., Pedersen A. K., Brash A. R. Increased prostacyclin biosynthesis in patients with severe atherosclerosis and platelet activation. N Engl J Med. 1984 Apr 26;310(17):1065–1068. doi: 10.1056/NEJM198404263101701. [DOI] [PubMed] [Google Scholar]
- Furchgott R. F., Zawadzki J. V. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 1980 Nov 27;288(5789):373–376. doi: 10.1038/288373a0. [DOI] [PubMed] [Google Scholar]
- Goldsmith J. C. Contribution of the subendothelium to prostacyclin release after vascular injury. J Lab Clin Med. 1982 Oct;100(4):574–584. [PubMed] [Google Scholar]
- Hajjar D. P., Falcone D. J., Fowler S., Minick C. R. Endothelium modifies the altered metabolism of the injured aortic wall. Am J Pathol. 1981 Jan;102(1):28–39. [PMC free article] [PubMed] [Google Scholar]
- Hornstra G., Haddeman E., Don J. A. Some investigations into the role of prostacyclin in thromboregulation. Thromb Res. 1978 Feb;12(2):367–374. doi: 10.1016/0049-3848(78)90308-0. [DOI] [PubMed] [Google Scholar]
- Ingerman-Wojenski C., Silver M. J., Smith J. B., Macarak E. Bovine endothelial cells in culture produce thromboxane as well as prostacyclin. J Clin Invest. 1981 May;67(5):1292–1296. doi: 10.1172/JCI110157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kent R. S., Diedrich S. L., Whorton A. R. Regulation of vascular prostaglandin synthesis by metabolites of arachidonic acid in perfused rabbit aorta. J Clin Invest. 1983 Aug;72(2):455–465. doi: 10.1172/JCI110993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Larrue J., Dorian B., Demond-Henri J., Bricaud H. Endogenous arachidonic acid metabolism by cultured arterial smooth muscle cells. Biochem Biophys Res Commun. 1981 Aug 14;101(3):861–867. doi: 10.1016/0006-291x(81)91829-5. [DOI] [PubMed] [Google Scholar]
- Larrue J., Rigaud M., Daret D., Demond J., Durand J., Bricaud H. Prostacyclin production by cultured smooth muscle cells from atherosclerotic rabbit aorta. Nature. 1980 Jun 12;285(5765):480–482. doi: 10.1038/285480a0. [DOI] [PubMed] [Google Scholar]
- MacIntyre D. E., Pearson J. D., Gordon J. L. Localisation and stimulation of prostacyclin production in vascular cells. Nature. 1978 Feb 9;271(5645):549–551. doi: 10.1038/271549a0. [DOI] [PubMed] [Google Scholar]
- Maclouf J. A radioimmunoassay for 6-keto-PGF1 alpha. Methods Enzymol. 1982;86:273–286. doi: 10.1016/0076-6879(82)86200-9. [DOI] [PubMed] [Google Scholar]
- Marnett L. J., Siedlik P. H., Fung L. W. Oxidation of phenidone and BW755C by prostaglandin endoperoxide synthetase. J Biol Chem. 1982 Jun 25;257(12):6957–6964. [PubMed] [Google Scholar]
- Morrison A. D., Orci L., Berwick L., Perrelet A., Winegrad A. I. The effects of anoxia on the morphology and composite metabolism of the intact aortic intima-media preparation. J Clin Invest. 1977 Jun;59(6):1027–1037. doi: 10.1172/JCI108725. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ody C., Seillan C., Russo-Marie F. 6-Ketoprostaglandin F1 alpha, prostaglandins E2, F2 alpha and thromboxane B2 production by endothelial cells, smooth muscle cells and fibroblasts cultured from piglet aorta. Biochim Biophys Acta. 1982 Jul 20;712(1):103–110. doi: 10.1016/0005-2760(82)90090-x. [DOI] [PubMed] [Google Scholar]
- POOLE J. C., SANDERS A. G., FLOREY H. W. The regeneration of aortic endothelium. J Pathol Bacteriol. 1958 Jan;75(1):133–143. doi: 10.1002/path.1700750116. [DOI] [PubMed] [Google Scholar]
- Rapoport R. M., Draznin M. B., Murad F. Endothelium-dependent relaxation in rat aorta may be mediated through cyclic GMP-dependent protein phosphorylation. Nature. 1983 Nov 10;306(5939):174–176. doi: 10.1038/306174a0. [DOI] [PubMed] [Google Scholar]
- Rapoport R. M., Murad F. Agonist-induced endothelium-dependent relaxation in rat thoracic aorta may be mediated through cGMP. Circ Res. 1983 Mar;52(3):352–357. doi: 10.1161/01.res.52.3.352. [DOI] [PubMed] [Google Scholar]
- Sevilla M. D., Neta P., Marnett L. J. Reaction of the antithrombotic and antimetastatic agent, nafazatrom, with oxidizing radicals. Biochem Biophys Res Commun. 1983 Sep 30;115(3):800–806. doi: 10.1016/s0006-291x(83)80005-9. [DOI] [PubMed] [Google Scholar]
- Siess W., Dray F., Seillan C., Ody C., Russo-Marie F. Prostanoid synthesis by vascular slices and cultured vascular cells of piglet aorta. Biochem Biophys Res Commun. 1981 Mar 31;99(2):608–616. doi: 10.1016/0006-291x(81)91788-5. [DOI] [PubMed] [Google Scholar]
- Smith W. L., DeWitt D. L., Allen M. L. Bimodal distribution of the prostaglandin I2 synthase antigen in smooth muscle cells. J Biol Chem. 1983 May 10;258(9):5922–5926. [PubMed] [Google Scholar]
- Stetz E. M., Majno G., Joris I. Cellular pathology of the rat aorta. Pseudo-vacuoles and myo-endothelial herniae. Virchows Arch A Pathol Anat Histol. 1979 Jul 31;383(2):135–148. doi: 10.1007/BF01200895. [DOI] [PubMed] [Google Scholar]
- Whiting J., Salata K., Bailey J. M. Aspirin: an unexpected side effect on prostacyclin synthesis in cultured vascular smooth muscle cells. Science. 1980 Nov 7;210(4470):663–665. doi: 10.1126/science.6776627. [DOI] [PubMed] [Google Scholar]
- Wickramasinghe J. A., Morozowich W., Hamlin W. E., Shaw S. R. Detection of prostaglandin F2 alpha as pentafluorobenzyl ester by electron-capture GLC. J Pharm Sci. 1973 Sep;62(9):1428–1431. doi: 10.1002/jps.2600620906. [DOI] [PubMed] [Google Scholar]
- Wight T. N., Curwen K. D., Litrenta M. M., Alonso D. R., Minick C. R. Effect of endothelium on glycosaminoglycan accumulation in injured rabbit aorta. Am J Pathol. 1983 Nov;113(2):156–164. [PMC free article] [PubMed] [Google Scholar]

