Cardiovascular Pharmacology

High density lipoprotein 3 inhibits oxidized low density lipoprotein-induced apoptosis via promoting cholesterol efflux in RAW264.7 cells



To investigate the protective effect of high density lipoprotein 3 (HDL3) on oxidized low density lipoprotein (ox-LDL)-induced apoptosis in RAW264.7 cells.


RAW264.7 cells were exposed to 50 mg/L ox-LDL for various durations up to 48 h, and apoptosis was detected using Hoechst 33258 staining and flow cytometric analysis. Total cholesterol levels were detected by high performance liquid chromatography, cholesterol efflux was determined by Tritium labeling, and the cellular lipid droplets were assayed by oil red O staining.


Treatment with 50 mg/L ox-LDL for 12, 24, and 48 h increased the apoptotic rate of RAW264. 7 cells in a time-dependent manner. The peak apoptotic rate (47.7%) was observed after 48 h incubation. HDL3 at various concentrations (50 mg/L, 100 mg/L, and 200 mg/L) inhibited the ox-LDL (50 mg/L for 48 h)-mediated apoptosis that was accompanied by an increased rate of intracellular cholesterol efflux, and decreased total cholesterol levels in cells in a concentration-dependent manner. Blockage of cholesterol efflux by brefeldin decreased the protective effect of HDL3 on ox-LDL induced apoptosis. Increase of the cholesterol efflux effected by another cholesterol acceptor, β-cyclodextrin, led to a dramatic decrease in the apoptotic rate of cells.


HDL3 antagonizes ox-LDL-induced apoptosis in RAW264.7 cells, through reducing the accumulation of toxic cholesterol.


  1. 1

    Littlewood TD, Bennet MR . Apoptotic cell death in atherosclerosis. Current Opin Lipidol 2003; 14: 469–75.

    CAS  Article  Google Scholar 

  2. 2

    Stoneman VE, Bennett MR . Role of apoptosis in atherosclerosis and its therapeutic implications. Clin Sci 2004; 107: 343–54.

    CAS  Article  Google Scholar 

  3. 3

    Kolodgie FD, Narula J, Haider N . Apoptosis in atherosclerosis. Does it contribute to plaque instability? Cardiol Clin 2001; 19: 127–39.

    CAS  Article  Google Scholar 

  4. 4

    Best PJM, Hasdai D, Sangiorgi G, Schwartz RS, Holmes DR, Simari RD, et al. Apoptosis Basic concepts and implication in coronary artery disease. Arterioscler Thromb Vasc Biol 1999; 19: 14–22.

    CAS  Article  Google Scholar 

  5. 5

    Kinscherf R, Claus R, Wagner M, Gehrke C, Kamencic H, Hou D, et al. Apoptosis caused by oxidized LDL is manganese superoxide dismutase and p53 dependent. FASEB J 1998; 12: 461–7.

    CAS  Article  Google Scholar 

  6. 6

    Wintergerst ES, Jelk J, Rahner C, Asmis R . Apoptosis induced by oxidized low density lipoprotein in human monocyte-derived macrophages involves CD36 and activation of caspase-3. Eur J Biochem 2000; 267: 6050–9.

    CAS  Article  Google Scholar 

  7. 7

    Toru K, Kume N, Minami M, Hayashida K, Murayama T, Sano H, et al. Role of oxidized LDL in atherosclerosis. Ann NY Acad Sci 2001; 947: 199–206.

    Google Scholar 

  8. 8

    Winther MP, Dijk KW, Havekes LM, Hofker MH . Macrophage scavenger receptor class a: a multifunctional receptor in atherosclerosis. Arterioscler Thromb Vasc Biol 2000; 20: 290–7.

    Article  Google Scholar 

  9. 9

    Dhaliwal BS, Steinbrecher UP . Cholesterol delivered to macrophages by oxidized low density lipoprotein is sequestered in lysosomes and fails to efflux normally. J Lipid Res 2000; 41: 1658–65.

    CAS  PubMed  Google Scholar 

  10. 10

    Yancey PG, Jerome WG . Lysosomal sequestration of free and esterified cholesterol from oxidized low density lipoprotein in macrophages of different species. J Lipid Res 1998; 39: 1349–61.

    CAS  PubMed  Google Scholar 

  11. 11

    Tabas I . Consequences of cellular cholesterol accumulation: basic concepts and physiological implications. J Clin Invest 2002; 110: 905–11.

    CAS  Article  Google Scholar 

  12. 12

    Weibel GK, Jerome WG, Small DM, Warner GJ, Stoltenborg JK, Kearney MA, et al. Effects of intracellular free cholesterol accumulation on macrophage viability: a model for foam cell death. Arterioscler Thromb Vasc Biol 1998; 18: 423–31.

    Article  Google Scholar 

  13. 13

    Yao PM, Tabas I . Free cholesterol loading of macrophages is associated with widespread mitochondrial dysfunction and activation of the mitochondrial apoptosis pathway. J Biol Chem 2001; 276: 42468–76.

    CAS  Article  Google Scholar 

  14. 14

    Yao PM, Tabas I . Free cholesterol loading of macrophages induces apoptosis involving the Fas pathway. J Biol Chem 2000; 275: 23807–13.

    CAS  Article  Google Scholar 

  15. 15

    Rader DJ . High-density lipoproteins and atherosclerosis. Am J Cardiol 2002; 90: 62–70.

    Article  Google Scholar 

  16. 16

    Eckardstein AV, Nofer JR, Assmann G . High-density lipoproteins and arteriosclerosis: role of cholesterol efflux and reverse cholesterol transport. Arterioscler Thromb Vasc Biol 2001; 21: 13–27.

    Article  Google Scholar 

  17. 17

    Sugano M, Tsuchida K, Makino N . High-density lipoproteins protect endothelial cells from tumor necrosis factor-alpha-induced apoptosis. Biochem Biophys Res Commun 2000; 272: 872–6.

    CAS  Article  Google Scholar 

  18. 18

    Yancey PG, Rodrigueza WV, Stoudt GW, Johnson WJ, Philips MC, Rothblat GH . Cellular cholesterol efflux mediated by cyclodextrins. J Biol Chem 1996; 271: 16026–34.

    CAS  Article  Google Scholar 

  19. 19

    Mendez AJ . Monensin and Brefeldin A inhibit high density lipoprotein-mediated cholesterol efflux from cholesterol-enriched cells. J Biol Chem 1995; 270: 5891–900.

    CAS  Article  Google Scholar 

  20. 20

    Li LX, Chen JX, Liao DF, Yu L . Probucol inhibits oxidized-low density lipoprotein induced adhesion of monocytes to endothelial cells by reducing p-selectin synthesis in vitro. Endothelium 1998; 6: 1–8.

    CAS  Article  Google Scholar 

  21. 21

    Cullen P, Fobker M, Tegelkamp K, Meyer K, Kannenberg F, Cignarella A, et al. An improved method for quantification of cholesterol and cholesteryl esters in human monocyte-derived macrophages by high performance liquid chromatography with identification of unassigned cholesteryl ester species by means of secondary ion mass spectrometry. J Lipid Res 1997; 38: 401–9.

    CAS  PubMed  Google Scholar 

  22. 22

    Nofer JR, Levkau B, Wolinska I, Junker R, Fobker M, von Eckardstein A, et al. Suppression of endothelial cell apoptosis by high density lipoproteins (HDL) and HDL-associated lysosphingolipids. J Biol Chem 2001; 276: 34480–5.

    CAS  Article  Google Scholar 

  23. 23

    Martinet W, Knaapen MW, De Meyer GR, Herman AG, Kockx MM . Oxidative DNA damage and repair in experimental atherosclerosis are reversed by dietary lipid lowering. Circ Res 2001; 88: 733–9.

    CAS  Article  Google Scholar 

  24. 24

    Rothblat GH, Atger V, Kellner-Weibel G, Williams DL, Phillips MC . Cell cholesterol efflux: integration of old and new observations provides new insights. J Lipid Res 1999; 40: 781–96.

    CAS  PubMed  Google Scholar 

  25. 25

    Rye KA, Clay MA, Barter PJ . Remodeling of high density lipoproteins by plasma factors. Atherosclerosis 1999; 145: 227–38.

    CAS  Article  Google Scholar 

  26. 26

    Attie A, Kastelein JP, Hayden MR . Pivotal role of ABCA1 in reverse cholesterol transport influencing HDL levels and susceptibility to atherosclerosis. J Lipid Res 2001; 42: 1717–26.

    CAS  PubMed  Google Scholar 

  27. 27

    Tall AR, Costet P, Wang N . Regulation and mechanisms of macrophage cholesterol efflux. J Clin Invest 2002; 110: 899–904.

    CAS  Article  Google Scholar 

  28. 28

    Oram JF, Albers JJ, Cheung MC, Bierman EL . The effects of subtractions of high density lipoprotein on cholesterol efflux from cultured fibroblasts Regulation of low density lipoprotein receptor activity. J Biol Chem 1981; 256: 8348–562

    CAS  PubMed  Google Scholar 

  29. 29

    Anatol K, Sandrine C, Chapman MJ . Small, dense HDL particles exert potent protection of atherogenic LDL against oxidative stress. Arterioscler Thromb Vasc Biol 2003; 23: 1881–8.

    Article  Google Scholar 

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Corresponding authors

Correspondence to Wei-dong Yin or Duan-fang Liao.

Additional information

Project supported by the National Natural Science Foundation of China (No 30171084, 30470719), the National Major Basic Research Program of Ministry of Science and Technology of China (973)(G2000056905) and the Natural Science Foundation of Hunan (01JJY2145).

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Jiang, P., Yan, P., Chen, J. et al. High density lipoprotein 3 inhibits oxidized low density lipoprotein-induced apoptosis via promoting cholesterol efflux in RAW264.7 cells. Acta Pharmacol Sin 27, 151–157 (2006).

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  • cholesterol efflux
  • high density lipoprotein-3
  • apoptosis
  • oxidized low density lipoprotein

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