Thromb Haemost 1992; 68(06): 744-751
DOI: 10.1055/s-0038-1646354
Original Article
Schattauer GmbH Stuttgart

Vitamin E Binds to Specific Binding Sites and Enhances Prostacyclin Production by Cultured Aortic Endothelial Cells

Makoto Kunisaki
The Third Department of Internal Medicine, Faculty of Medicine, Kyushu University Fukuoka, Japan
,
Fumio Umeda
The Third Department of Internal Medicine, Faculty of Medicine, Kyushu University Fukuoka, Japan
,
Toyoshi Inoguchi
The Third Department of Internal Medicine, Faculty of Medicine, Kyushu University Fukuoka, Japan
,
Hajime Nawata
The Third Department of Internal Medicine, Faculty of Medicine, Kyushu University Fukuoka, Japan
› Author Affiliations
Further Information

Publication History

Received 26 June 1991

Accepted after revision 22 July 1992

Publication Date:
04 July 2018 (online)

Summary

We evaluated the effect of d-α-tocopherol (vitamin E) on the production of prostacyclin (PGI2) by cultured bovine aortic endothelial cells. Vitamin E at physiological doses significantly enhanced the production of PGI2 by aortic endothelial cells when added to the culture simultaneously with histamine, the Ca2+ ionophore A23187 (A23187), plasma-derived serum (PDS), or arachidonic acid. This effect was found to occur in a time- and dose-dependent manner, and the maximal enhancement was produced by 9.28 µM of vitamin E for 1 h incubations. Significantly lower amounts of lipid peroxides were measured in endothelial cells stimulated by 10% PDS with 9.28 µM of vitamin E than in those stimulated without vitamin E for over 24 h, although the stimulation during the initial 1 to 12 h period did not have a significant effect on lipid peroxide formation in cultured aortic endothelial cells.

We also demonstrated that bovine aortic endothelial cells have specific binding sites for [3H]vitamin E that exhibited time- and temperature-dependent saturability. At 4° C, the nonspecific binding was 8–12% of the total binding, and the specific binding reached equilibrium by 2 h. Specific binding increased with the concentration of [3H]vitamin E and became saturated at concentrations between 1.5 µM and 2.0 µM per 2.0 × 105 cells. Raising the unlabeled vitamin E concentration from 97.7 nM to 1,000 µM reduced the specific binding of 2.0 µM [3H]vitamin E. The Scatchard plot of [3H]vitamin E binding to the endothelial cells shows two classes of binding sites: one with a high affinity {K a1 2.48 ± 0.32 × 107 NT-1, n = 6} and a low capacity {n 1 1.20 ± 0.34 × 107 sites/cell} and the other with a low affinity {K a2 1.18 ± 0.32 × 105 M–1} and a high capacity {n 2 3.39 ± 0.53 × 109 sites/cell}.

Our results suggest that the endothelial cells binding sites for vitamin E may play some roles in vascular homeostasis in vivo, and that vitamin E may prevent the development of atherosclerotic changes due in part to the enhancement of PGI2 production by the vascular wall and its action as an antioxidant in vascular endothelial cell.

 
  • REFERENCES

  • 1 Tappel AL. Vitamin E and free radical peroxidation of lipids. Ann N Y Acad Sci 1976; 203: 12-28
  • 2 Bieri JG, Farrell PM. Vitamin E. Vitamins Horm 1976; 34: 31-75
  • 3 Baehner RL, Boxer LA, Allen JM, Davis J. Autooxidation as a basis for altered function by polymorphonuclear leukocytes. Blood 1977; 50: 327-335
  • 4 Kitabchi AE. Hormonal status in vitamin E deficiency. In: Vitamin E. Machlin LJ. (ed) Marcel Dekker Inc., New York: 1980. pp 348-371
  • 5 Moncada S, Gryglewski RJ, Bunting S, Vane JR. An enzyme isolated from arteries transforms prostaglandin endoperoxides to an unstable substance that inhibits platelet aggregation. Nature 1976; 263: 663-665
  • 6 Moncada S, Vane JR. Unstable metabolites of arachidonic acid and their role in haemostasis and thrombosis. Br Med Bull 1978; 34: 129-135
  • 7 Moncada S, Higgs EA, Vane JR. Human arterial and venous tissues generate prostacyclin (Prostaglandin X), a potent inhibitor of platelet aggregation. Lancet i 1977; 18-20
  • 8 Moncada S, Gryglewski RJ, Bunting S, Vane JR. A lipid peroxide inhibits the enzyme in blood vessel microsomes that generates from prostaglandin endoperoxides the substance (prostaglandin X) which prevents platelet aggregation. Prostaglandins 1976; 12: 715-737
  • 9 Gryglewski RJ, Bunting S, Moncada S, Flower RJ, Vane JR. Arterial walls are protected against deposition of platelet thrombi by a substance (prostaglandin X) which they make from prostaglandin endoperoxides. Prostaglandins 1976; 12: 685-713
  • 10 McIntyre DD, Pearson JD, Gordon JL. Localization and stimulation of prostacyclin production in vascular cells. Nature 1978; 271: 549-551
  • 11 Hong SL, Carty T, Deykin D. Tranylcypromine and 15-hydroperoxy arachidonate affect arachidonic acid release in addition to inhibition of prostacyclin synthesis in calf aortic endothelial cells. J Biol Chem 1980; 255: 9538-9540
  • 12 Okuma M, Takayama H, Uchino H. Generation of prostacyclin-like substance and lipid peroxidation in vitamin E-deficient rats. Prostaglandins 1980; 19: 527-536
  • 13 Chan AC, Leith MK. Decreased prostacyclin synthesis in vitamin E-deficient rabbit aorta. Am J Clin Nutr 1981; 34: 2341-2347
  • 14 Sugimoto H, Matumoto M, Kimura T, Ibayashi H. The effect of vitamin E on the prostaglandins metabolism: Plasma lipoperoxide and aortic prostacyclin production in vitamin E deficient rats. Jpn Athero Soc 1980; 8: 575-578
  • 15 Kunisaki M, Umeda F, Inoguchi T, Ono H, Sako Y. Effect of vitamin E on prostacyclin production from cultured aortic endothelial cells. In: Endothelium in Health and Disease. Chai S. (ed) Plenum Publishing Co., New York: 1988. pp 113-118
  • 16 Inoguchi T, Umeda F, Watanabe J, Ibayashi H. Reduced serum stimulatory activity on prostacyclin production by cultured aortic endothelial cells in diabetes mellitus. Haemostasis 1986; 16: 447-452
  • 17 Ono H, Umeda F, Inoguchi T, Nawata H. Glucose inhibits prostacyclin production by cultured aortic endothelial cells. Thromb Haemostas 1988; 60: 174-177
  • 18 Yamauchi T, Onaka K, Ryoichi T, Umeda F, Nawata H. Enhanced secretion of endothelin-I by elevated glucose levels from cultured bovine aortic endothelial cells. FEBS Lett 1990; 267: 16-18
  • 19 Weksler BB, Ley CW, Jaffe EA. Stimulation of endothelial cell prostacyclin production by thrombin, trypsin, and the ionophore A23187. J Clin Invest 1978; 62: 923-930
  • 20 Alhence-Gelas F, Tsai SJ, Callahan KS, Campbell WB, Johnson AE. Stimulation of prostaglandin formation by vasoactive mediators in cultured human endothelial cells. Prostaglandins 1982; 24: 723-742
  • 21 Pledger WJ, Stiles CD, Antoniades HN, Scher CD. Induction of DNA synthesis in BALB/c 3T3 cells by serum components: revaluation of the commitment process. Proc Natl Acad Sci USA 1977; 74: 4481-4485
  • 22 Umeda F, Franks DJ, Sugimoto H, Halle JP, Hamet P. Effect of plasma, serum and platelets from diabetics on DNA synthesis in cultured vascular smooth muscle cells. Clin Invest Med 1985; 8: 35-40
  • 23 Jaffe BM, Behrman HR, Parker CW. Radioimmunoassay measurement of prostaglandins E, A, and F in human plasma. J Clin Invest 1973; 52: 398-405
  • 24 Yagi K. A simple fluorometric assay for lipoperoxide in blood plasma. Biochem Med 1976; 15: 212-216
  • 25 Scatchard G. The attraction of proteins for small molecules and ions. Ann N Y Acad Sci 1949; 51: 660-675
  • 26 Thakur AK, Jaffe ML, Rodbard D. Graphical analysis of ligand-binding system: evaluation by Monte Carlo Studies. Analyt Biochem 1980; 107: 279-295
  • 27 Huang N, Lineberger B, Stiner M. Alpha-tocopherol, a potent modulator of endothelial cell function. Thromb Res 1988; 50: 547-557
  • 28 Toivanen JL. Effect of selenium, vitamin E and vitamin C on human prostacyclin and thromboxane synthesis in vitro. Prostaglandins Leukotriens Med 1987; 26: 265-280
  • 29 Hopper K, Phillips WEJ, Murray TK, Campbell JS. Data on serum tocopherol levels in a selected group of Canadians. Can J Physiol Pharmacol 1970; 48: 321-323
  • 30 Bokman SO, Larsson C. The distribution of prostaglandin synthetase in membrane fractions from the rabbit renal medulla. J Urtrastr Res 1975; 50: 367
  • 31 Rollins TE, Smith WL. Subcellular localization of prostaglandin-forming cyclooxygenase in Swiss mouse 3T3 fibroblasts by electron microscopic immunocytochemistry. J Biol Chem 1980; 1.0: 4872-4875
  • 32 Willems C, De Groot PG, Pool GA, Gonsalvez MS, Van Aken WG, Van Mourik JA. Arachidonate metabolism in cultured human vascular endothelial cells. Evidence for two prostaglandin synthetic pathways sensitive to acetylsalicylic acid. Biochim Biophys Acta 1982; 713: 581-588
  • 33 Gisinger C, Watanabe J, Colwell JA. Vitamin E and platelet eicosanoids in diabetes mellitus. Prostaglandins Leukotrienes Essential Fatty Acids 1990; 40: 169-176
  • 34 Kitabchi AE, Wimalasena J, Barker J. Specific receptor sites for α-tocopherol in purified isolated adrenocortical cell membrane. Biochem Biophys Res Comm 1980; 96: 1739-1746
  • 35 Kitabchi AE, Wimalasena J. Specific binding site for d-α-tocopherol on human erythrocyte. Biochim Biophys Acta 1982; 684: 200-206