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Enrichment of LDL with EPA and DHA decreased oxidized LDL-induced apoptosis in U937 cells

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Lipids

Abstract

Oxidized LDL (oxLDL) may contribute to the accumulation of apoptotic cells in atherosclerotic plaques. Although it is well established in monophasic chemical systems that the highly unsaturated EPA and DHA will oxidize more readily than FA that contain fewer double bonds, our previous studies showed that enrichment of LDL, which has discrete polar and nonpolar phases, with these FA did not increase oxidation. The objective of this study was to compare the extent of apoptosis induced by EPA/DHA-rich oxLDL to that induced by EPA/DHA-non-rich oxLDL in U937 cells. LDL was obtained from one healthy subject three times before and after supplementation for 5 wk with 15 g/d of fish oil (FO), an amount easily obtainable from a diet that contains fatty fish. After supplementation, an FPA/DHA-rich LDL was obtained. Oxidative susceptibility of LDL, as determined by measuring the formation of conjugated dienes and the accumulation of cholesteryl ester hydroperoxides, was not higher in EPA/DHA-rich LDL. The oxLDL-induced cell apoptosis was detected by the activation of caspase-3, the translocation of PS to the outer surface of the plasma membrane using the Annexin V-fluorescein isothiocyanate binding assay, and the presence of chromatin condensation and nuclear fragmentation using the 4,6-diamidino-2-phenylindole staining assay All three measures showed that after FO supplementation, EPA/DHA-rich oxLDL-induced cell apoptosis decreased. The decrease was not related to the concentration of lipid hydroperoxides. This study suggests that a possible protective effect of EPA/DHA-rich diets on atherosclerosis may be through lessening cell apoptosis in the arterial wall.

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Abbreviations

AMC:

aminomethyl coumarin

CE18∶200H:

cholesteryl linoleate hydroperoxide

CEOOH:

cholesteryl ester hydroperoxides

DAPI:

4,6-diamidino-2-phenylindole

FITC:

fluorescein isothiocyanate

FO:

fish oil

HDL-C:

HDL-cholesterol

LDL-C:

LDL-cholesterol

nLDL:

native LDL

oxLDL:

oxidized low-density lipoprotein

PI:

propidium iodide

References

  1. Stehbens, W.E. (2000) The Significance of Programmed Cell Death or Apoptosis and Matrix Vesicles in Atherogenesis, Cell. Mol. Biol. 46, 99–110.

    PubMed  CAS  Google Scholar 

  2. Björkerud, S., and Björkerud, B. (1996) Apoptosis Is Abundant in Human Atherosclerotic Lesions, Especially in Inflammatory Cells (macrophages and T cells), and May Contribute to the Accumulation of Gruel and Plaque Instability, Am. J. Pathol. 149, 367–380.

    PubMed  Google Scholar 

  3. Han, D.K.M., Haudenschild, C.C., Hong, M.K., Tinkle, B.T., Leon, M.B., and Liau, G. (1995) Evidence for Apoptosis in Human Atherogenesis and in a Rat Vascular Injury Model, Am. J. Pathol. 147, 267–277.

    PubMed  CAS  Google Scholar 

  4. Mitchinson, M.J., Hardwick, S.J., and Bennett, M.R. (1996) Cell Death in Atherosclerotic Plaques, Curr. Opin. Lipidol. 7, 324–329.

    Article  PubMed  CAS  Google Scholar 

  5. Hegyi, L., Skepper, J.N., Cary, N.R., and Mitchinson, M.J. (1996) Foam Cell Apoptosis and the Development of the Lipid Core of Human Atherosclerosis, J. Pathol. 180, 423–429.

    Article  PubMed  CAS  Google Scholar 

  6. Isner, J.M., Kearney, M., Bortman, S., and Passeri, J. (1995) Apoptosis in Human Atherosclerosis and Restenosis, Circulation 91, 2703–2711.

    PubMed  CAS  Google Scholar 

  7. Ball, R.Y., Stowers, E.C., Burton, J.H., Cary, N.R., Skepper, J.N., and Mitchinson, M.J. (1995) Evidence That the Death of Macrophage Foam Cells Contributes to the Lipid Core of Atheroma, Atherosclerosis 114, 45–54.

    Article  PubMed  CAS  Google Scholar 

  8. Hegyi, L., Hardwick, S.J., Siow, R.C., and Skepper, J.N. (2001) Macrophage Death and the Role of Apoptosis in Human Atherosclerosis, J. Hematother. Stem Cell. Res. 10, 27–42.

    Article  PubMed  CAS  Google Scholar 

  9. Kockx, M.M., and Knaapen, M.W. (2000) The Role of Apoptosis in Vascular Disease, J. Pathol. 190, 267–280.

    Article  PubMed  CAS  Google Scholar 

  10. Witztum, J.L., and Steinberg, D. (1991) Role of Oxidized LDL in Atherogenesis, J. Clin. Invest. 88, 1785–1792.

    PubMed  CAS  Google Scholar 

  11. Berliner, J.A., and Heinecke, J.W. (1996) The Role of Oxidized Lipoproteins in Atherosclerosis, Free Radic. Biol. Med. 20, 707–727.

    Article  PubMed  CAS  Google Scholar 

  12. Dimmeler, S., Haendeler, J., Galle, J., and Zeiher, A.M. (1997) Oxidized Low-Density Lipoprotein Induces Apoptosis of Human Endothelial Cells by Activation of CPP32-like Proteases. A Mechanistic Clue to the ‘Response to Injury’ Hypothesis, Circulation 95, 1760–1763.

    PubMed  CAS  Google Scholar 

  13. Harada-Shiba, M.K., Kinoshita, M.K., Kamido, H.S., and Shimokado, K.T. (1998) Oxidized Low Density Lipoprotein Induces Apoptosis in Cultured Human Umbilical Vein Endothelial Cells by Common and Unique Mechanisms, J. Biol. Chem. 273, 9681–9687.

    Article  PubMed  CAS  Google Scholar 

  14. Siow, R.C., Richards, J.P., Pedley, K.C., Leake, D.S., and Mann, G.E. (1999) Vitamin C Protects Human Vascular Smooth Muscle Cells Against Apoptosis Induced by Moderately Oxidized LDL Containing High Levels of Lipid Hydroperoxides, Arterioscler. Thromb. Vasc. Biol. 19, 2387–2394.

    CAS  Google Scholar 

  15. Wintergerst, E.S., Jelk, J., Rahner, C., and Asmis, R. (2000) Apoptosis Induced by Oxidized Low Density Lipoprotein in Human Monocyte-Derived Macrophages Involves CD36 and Activation of Caspase-3, Eur. J. Biochem. 267, 6050–6059.

    Article  PubMed  CAS  Google Scholar 

  16. Hardwick, S.J., Hegyi, L., Clare, K., Law, N.S., Carpenter, K.L., Mitchinson, M.J., and Skepper, J.N. (1996) Apoptosis in Human Monocyte-Macrophages Exposed to Oxidized Low Density Lipoprotein, J. Pathol. 179, 294–302.

    Article  PubMed  CAS  Google Scholar 

  17. Wang, T.G., Gotoh, Y., Jennings, M.H., Rhoads, C.A., and Aw, T.Y. (2000) Lipid Hydroperoxide-Induced Apoptosis in Human Colonic CaCo-2 Cells Is Associated with an Early Loss of Cellular Redox Balance, FASEB J. 14, 1567–1576.

    Article  PubMed  CAS  Google Scholar 

  18. Liu, W., Kato, M., Akhand, A.A., Hayakawa, A., Suzuki, H., Miyata, T., Kurokawa, K., Hotta, Y., Ishikawa, N., and Nakashima, I. (2000) 4-Hydroxynonenal Induces a Cellular Redox Status-Related Activation of the Caspase Cascade for Apoptotic Cell Death, J. Cell. Sci. 113, 635–641.

    PubMed  CAS  Google Scholar 

  19. Lizard, G., Monier, S., Cordelet, C., Gesquiere, L., Deckert, V., Gueldry, S., Lagrost, L., and Gambert, P. (1999) Characterization and Comparison of the Mode of Cell Death, Apoptosis Versus Necrosis, Induced by 7β-Hydroxycholesterol and 7-Ketocholesterol in the Cells of the Vascular Wall, Arterioscler. Thromb. Vasc. Biol. 19, 1190–2000.

    PubMed  CAS  Google Scholar 

  20. Spyridopoulos, I., Wischhusen, J., Rabenstein, B., Mayer, P., Axel, D.I., Frohlich, K.U., and Karsch, K.R. (2001) Alcohol Enhances Oxysterol-Induced Apoptosis in Human Endothelial Cells by a Calcium-Dependent Mechanism, Arterioscler. Thromb. Vasc. Biol. 21, 439–444.

    PubMed  CAS  Google Scholar 

  21. Yang, L., and Sinensky, M.S. (2000) 25-Hydroxycholesterol Activates a Cytochrome c Release-Mediated Caspase Cascade, Biochem. Biophys. Res. Commun. 278, 557–563.

    Article  PubMed  CAS  Google Scholar 

  22. Lee, T., and Chau, L. (2001) Fas/Fas Ligand-Mediated Death Pathway Is Involved in OxLDL-Induced Apoptosis in Vascular Smooth Muscle Cells, Am. J. Cell. Physiol. 280, C709-C718.

    CAS  Google Scholar 

  23. Vieira, O., Escargueil-Blanc, I., Meilhac, O., Basile, J.P., Laranjinha, J., Almeida, L., Salvayre, R., and Negre-Salvayre, A. (1998) Effect of Dietary Phenolic Compounds on Apoptosis of Human Cultured Endothelial Cells Induced by Oxidized LDL. Br. J. Pharmacol. 123: 565–573.

    Article  PubMed  CAS  Google Scholar 

  24. Higdon, J.V., Du, S.H., Lee, Y.S., Wu, T., and Wander, R.C. (2001) Supplementation of Postmenopausal Women with Fish Oil Does Not Increase Overall Oxidation of LDL, ex vivo Compared to Dietary Oils Rich in Oleate and Linoleate, J. Lipid Res. 42, 407–418.

    PubMed  CAS  Google Scholar 

  25. Suarna, C., Dean, R.T., May, J., and Stocker, R. (1995) Human Atherosclerotic Plaque Contains Both Oxidized Lipids and Relatively Large Amounts of α-Tocopherol and Ascorbate, Arterioscler. Thromb. Vasc. Biol. 15, 1616–1624.

    PubMed  CAS  Google Scholar 

  26. Song, J., and Wander, R.C. (1991) Effects of Dietary Selenium and Fish Oil (MaxEPA) on Arachidonic Acid Metabolism and Hemostatic Function in the Rat, J. Nutr. 121, 284–292.

    PubMed  CAS  Google Scholar 

  27. Wander, R.C., and Patton, B.D. (1991) Lipids and Fatty Acids of Three Species of Northeast Pacific Finfish Harvested in Summer, Food Composition Anal. 4, 128–135.

    Article  CAS  Google Scholar 

  28. Arnaud, J., Fortis, I., Blachier, S., Kia, D., and Favier, A. (1991) Simultaneous Determination of Retinol, α-Tocopherol and β-Carotene in Serum by Isocratic High-Performance Liquid Chromatography, J. Chromatogr. 572, 103–116.

    PubMed  CAS  Google Scholar 

  29. Wander, R.C., Du, S.H., and Thomas, D.R. (1998) Influence of Long-Chain Polyunsaturated Fatty Acids on Oxidation of Low Density Lipoprotein, Prostaglandins Leukotrienes Essent. Fatty Acids 59, 143–151.

    Article  CAS  Google Scholar 

  30. Marz, W., and Gross, W. (1986) Analysis of Plasma Lipoproteins by Ultracentrifugation in a New Fixed Angle Rotor: Evaluation of a Phosphotungstic Acid/MgCl2 Precipitation and a Quantitative Lipoprotein Electrophoresis Assay, Clin. Chim. Acta 160, 1–18.

    Article  PubMed  CAS  Google Scholar 

  31. Friedewald, W.T., Levy, R.I., and Fredrickson, D.S. (1972) Estimation of the Concentration of Low-Density Lipoprotein Cholesterol in Plasma, Without Use of the Preparative Ultracentrifuge, Clin. Chem. 18, 499–502.

    PubMed  CAS  Google Scholar 

  32. Chung, B.H., Segrest, J.P., Ray, M.J., Brunzell, J.D., Hokanson, J.E., Krauss, R.M., Beaudrie, K., and Cone, J.T. (1986) Single Vertical Spin Density Gradient Ultracentrifugation, Methods Enzymol. 128, 181–209.

    Article  PubMed  CAS  Google Scholar 

  33. Chen, C., and Loo, G. (1995) Cigarette Smoke Extract Inhibits Oxidative Modification of Low Density Lipoprotein, Atherosclerosis 112, 177–185.

    Article  PubMed  CAS  Google Scholar 

  34. Wander, R.C., Du, S.-H., Ketchum, S.O., and Rowe, K.E. (1996) Interaction of Vitamin E and Fish Oil on LDL Oxidation in Post-menopausal Women With and Without Hormone Replacement Therapy, Am. J. Clin. Nutr. 63, 184–193.

    PubMed  CAS  Google Scholar 

  35. Esterbauer, H., Striegl, G., Puhl, H., and Rotheneder, M. (1989) Continuous Monitoring of in vitro Oxidation of Human Low Density Lipoprotein, Free Radic. Res. Commun. 6, 67–75.

    PubMed  CAS  Google Scholar 

  36. Stennicke, H.R., and Salvesen, G.S. (1997) Biochemical Characteristics of Caspases-3, −6, −7, and −8, J. Biol. Chem. 272, 25719–25723.

    Article  PubMed  CAS  Google Scholar 

  37. Martin, S.J., Reutelingsperger, C.P., McGahon, A.J., Rader, J.A., van Schie, R.C., LaFace, D.M., and Green, D.R. (1995) Early Redistribution of Plasma Membrane Phosphatidylserine Is a General Feature of Apoptosis Regardless of the Initiating Stimulus: Inhibition by Overexpression of Bcl-2 and Abl, J. Exp. Med. 182, 1545–1556.

    Article  PubMed  CAS  Google Scholar 

  38. Ramsey, F., and Schafer, D. (1999) Statistical Sleuth: A Course in Methods of Data Analysis, Duxbury Press, Boston.

    Google Scholar 

  39. Kuida, K., Haydar, T.F., Kuan, C.Y., Gu, Y., Taya, C., Karasuyama, H., Su, M.S., Rakic, P., and Flavell, R.A. (1998) Reduced Apoptosis and Cytochrome c-Mediated Caspase Activation in Mice Lacking Caspase 9, Cell 94, 325–337.

    Article  PubMed  CAS  Google Scholar 

  40. Song, Z., and Steller, H. (1999) Death by Design: Mechanism and Control of Apoptosis, Trends Cell Biol. 9, M49-M52.

    Article  PubMed  CAS  Google Scholar 

  41. Belloc, F., Belaud-Rotureau, M.A., Lavignolle, V., Bascans, E., Braz-Pereira, E., Durrieu, F., and Lacombe, F. (2000) Flow Cytometry Detection of Caspase 3 Activation in Prepoptotic Leukemic Cells, Cytometry 40, 151–160.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Human Nutrition Research Laboratory, University of North Carolina at Greensboro, P.O. Box 26170, 27402-6170, Greensboro, North Carolina

    Tianying Wu, Cissy Geigerman, Ye-Sun Lee & Rosemary C. Wander

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  1. Tianying Wu
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  2. Cissy Geigerman
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Correspondence to Rosemary C. Wander.

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Wu, T., Geigerman, C., Lee, YS. et al. Enrichment of LDL with EPA and DHA decreased oxidized LDL-induced apoptosis in U937 cells. Lipids 37, 789–796 (2002). https://doi.org/10.1007/s11745-002-0962-7

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  • DOI: https://doi.org/10.1007/s11745-002-0962-7

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