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Basic Research in Cardiology

, 106:761 | Cite as

OxLDL and macrophage survival: essential and oxygen-independent involvement of the Hif-pathway

  • David M. PoitzEmail author
  • Antje Augstein
  • Sönke Weinert
  • Rüdiger C. Braun-Dullaeus
  • Ruth H. Strasser
  • Alexander Schmeisser
Original Contribution

Abstract

Atherosclerotic plaques are characterized by hypoxic even anoxic areas and by high concentrations of oxidized lipoproteins. Moreover, unstable plaques attract a high number of macrophages despite the proapoptotic background within these plaques. Recently, it was shown that these macrophages are positive for Hif-1α. This subunit is a part of hypoxia-inducible factor 1 (Hif-1), a key transcriptional factor under hypoxia. Till date, it is not understood whether the Hif-system (consisting of Hif-1, Hif-2 and Hif-3) is involved in protection of macrophages under these proatherogenic conditions. The present study delineates that oxLDL causes fundamental changes in the regulation of the Hif-system in primary human macrophages. First, both oxLDL and hypoxia mediate accumulation of Hif-1α protein. Second, treatment with a combination of oxLDL and hypoxia is acting in an additive manner on Hif-1α protein content. Third, oxLDL alone does not increase Hif-2α protein, but abolishes the hypoxic induction of Hif-2α completely. OxLDL treatment alone was not toxic for macrophages under neither normoxia nor hypoxia. But, inhibition of Hif-pathway by adenoviral expression of a dominant-negative mutant combined with oxLDL treatment independently of the oxygen tension leads to apoptosis, as determined by caspase-3 activation and induction of DNA fragmentation. Furthermore, this inhibition also mediates the opening of the mitochondrial permeability transition pore. In conclusion, the present data show that Hif-1α regulation is essential for survival of oxLDL-treated macrophages independent of the oxygen tension. Therefore, this newly characterized mechanism might also have an important influence for the vulnerability of atherosclerotic plaques.

Keywords

Macrophages OxLDL Hypoxia-inducible factors Apoptosis 

Notes

Acknowledgments

R.B.-D. was supported by grants of the SFB 655. This project was also supported by grants of the MedDrive program and the CRTD. Anita Männel and Peggy Barthel are acknowledged for excellent technical assistance.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Bjornheden T, Levin M, Evaldsson M, Wiklund O (1999) Evidence of hypoxic areas within the arterial wall in vivo. Arterioscler Thromb Vasc Biol 19:870–876PubMedCrossRefGoogle Scholar
  2. 2.
    Blanc-Brude OP, Teissier E, Castier Y, Leseche G, Bijnens AP, Daemen M, Staels B, Mallat Z, Tedgui A (2007) IAP survivin regulates atherosclerotic macrophage survival. Arterioscler Thromb Vasc Biol 27:901–907PubMedCrossRefGoogle Scholar
  3. 3.
    Blouin CC, Page EL, Soucy GM, Richard DE (2004) Hypoxic gene activation by lipopolysaccharide in macrophages: implication of hypoxia-inducible factor 1α. Blood 103:1124–1130. doi: 10.1182/blood-2003-07-2427 PubMedCrossRefGoogle Scholar
  4. 4.
    Boullier A, Li Y, Quehenberger O, Palinski W, Tabas I, Witztum JL, Miller YI (2006) Minimally oxidized LDL offsets the apoptotic effects of extensively oxidized LDL and free cholesterol in macrophages. Arterioscler Thromb Vasc Biol 26:1169–1176. doi: 10.1161/01.ATV.0000210279.97308.9a PubMedCrossRefGoogle Scholar
  5. 5.
    Bracken CP, Whitelaw ML, Peet DJ (2003) The hypoxia-inducible factors: key transcriptional regulators of hypoxic responses. Cell Mol Life Sci 60:1376–1393. doi: 10.1007/s00018-003-2370-y PubMedCrossRefGoogle Scholar
  6. 6.
    Chen JH, Riazy M, Smith EM, Proud CG, Steinbrecher UP, Duronio V (2009) Oxidized LDL-mediated macrophage survival involves elongation factor-2 kinase. Arterioscler Thromb Vasc Biol 29:92–98. doi: 10.1161/ATVBAHA.108.174599 PubMedCrossRefGoogle Scholar
  7. 7.
    de Nigris F, Gallo L, Sica V, Napoli C (2006) Glycoxidation of low-density lipoprotein promotes multiple apoptotic pathways and NFkB activation in human coronary cells. Basic Res Cardiol 101:101–108. doi: 10.1007/s00395-005-0560-5 PubMedCrossRefGoogle Scholar
  8. 8.
    Dery MA, Michaud MD, Richard DE (2005) Hypoxia-inducible factor 1: regulation by hypoxic and non-hypoxic activators. Int J Biochem Cell Biol 37:535–540. doi: 10.1016/j.biocel.2004.08.012 PubMedCrossRefGoogle Scholar
  9. 9.
    Erbel C, Dengler TJ, Wangler S, Lasitschka F, Bea F, Wambsganss N, Hakimi M, Bockler D, Katus HA, Gleissner CA (2011) Expression of IL-17A in human atherosclerotic lesions is associated with increased inflammation and plaque vulnerability. Basic Res Cardiol 106:125–134. doi: 10.1007/s00395-010-0135-y PubMedCrossRefGoogle Scholar
  10. 10.
    Ermak N, Lacour B, Drueke TB, Vicca S (2008) Role of reactive oxygen species and Bax in oxidized low density lipoprotein-induced apoptosis of human monocytes. Atherosclerosis 200:247–256. doi: 10.1016/j.atherosclerosis.2007.12.052 PubMedCrossRefGoogle Scholar
  11. 11.
    Frede S, Stockmann C, Freitag P, Fandrey J (2006) Bacterial lipopolysaccharide induces HIF-1 activation in human monocytes via p44/42 MAPK and NF-kB. Biochem J 396:517–527. doi: 10.1042/BJ20051839 PubMedCrossRefGoogle Scholar
  12. 12.
    Fulda S, Debatin KM (2007) HIF-1-regulated glucose metabolism: a key to apoptosis resistance? Cell Cycle 6:790–792. doi: 10.4161/cc.6.7.4084 PubMedCrossRefGoogle Scholar
  13. 13.
    Garedew A, Henderson SO, Moncada S (2010) Activated macrophages utilize glycolytic ATP to maintain mitochondrial membrane potential and prevent apoptotic cell death. Cell Death Differ 17:1540–1550. doi: 10.1038/cdd.2010.27 PubMedCrossRefGoogle Scholar
  14. 14.
    Garedew A, Moncada S (2008) Mitochondrial dysfunction and HIF1α stabilization in inflammation. J Cell Sci 121:3468–3475. doi: 10.1242/jcs.034660 PubMedCrossRefGoogle Scholar
  15. 15.
    Ginouves A, Ilc K, Macias N, Pouyssegur J, Berra E (2008) PHDs overactivation during chronic hypoxia “desensitizes” HIFalpha and protects cells from necrosis. Proc Natl Acad Sci USA 105:4745–4750PubMedCrossRefGoogle Scholar
  16. 16.
    Gössl M, Herrmann J, Tang H, Versari D, Galili O, Mannheim D, Rajkumar SV, Lerman LO, Lerman A (2009) Prevention of vasa vasorum neovascularization attenuates early neointima formation in experimental hypercholesterolemia. Basic Res Cardiol 104:695–706. doi: 10.1007/s00395-009-0036-0 PubMedCrossRefGoogle Scholar
  17. 17.
    Jiang G, Li T, Qiu Y, Rui Y, Chen W, Lou Y (2007) RNA interference for HIF-1alpha inhibits foam cells formation in vitro. Eur J Pharmacol 562:183–190. doi: 10.1016/j.ejphar.2007.01.066 PubMedCrossRefGoogle Scholar
  18. 18.
    Kim HY, Kim YH, Nam BH, Kong HJ, Kim HH, Kim YJ, An WG, Cheong J (2007) HIF-1α expression in response to lipopolysaccaride mediates induction of hepatic inflammatory cytokine TNFa. Exp Cell Res 313:1866–1876. doi: 10.1016/j.yexcr.2007.03.009 PubMedCrossRefGoogle Scholar
  19. 19.
    Kohlstedt K, Trouvain C, Namgaladze D, Fleming I (2011) Adipocyte-derived lipids increase angiotensin-converting enzyme (ACE) expression and modulate macrophage phenotype. Basic Res Cardiol 106:205–215. doi: 10.1007/s00395-010-0137-9 PubMedCrossRefGoogle Scholar
  20. 20.
    Lall H, Coughlan K, Sumbayev VV (2008) HIF-1a protein is an essential factor for protection of myeloid cells against LPS-induced depletion of ATP and apoptosis that supports Toll-like receptor 4-mediated production of IL-6. Mol Immunol 45:3045–3049. doi: 10.1016/j.molimm.2008.03.014 PubMedCrossRefGoogle Scholar
  21. 21.
    Laughner E, Taghavi P, Chiles K, Mahon PC, Semenza GL (2001) HER2 (neu) signaling increases the rate of hypoxia-inducible factor 1α (HIF-1α) synthesis: novel mechanism for HIF-1-mediated vascular endothelial growth factor expression. Mol Cell Biol 21:3995–4004. doi: 10.1128/MCB.21.12.3995-4004.2001 PubMedCrossRefGoogle Scholar
  22. 22.
    Libby P (2002) Inflammation in atherosclerosis. Nature 420:868–874. doi: 10.1038/nature01323 PubMedCrossRefGoogle Scholar
  23. 23.
    Maemura K, Hsieh CM, Jain MK, Fukumoto S, Layne MD, Liu Y, Kourembanas S, Yet SF, Perrella MA, Lee ME (1999) Generation of a dominant-negative mutant of endothelial PAS domain protein 1 by deletion of a potent C-terminal transactivation domain. J Biol Chem 274:31565–31570. doi: 10.1074/jbc.274.44.31565 PubMedCrossRefGoogle Scholar
  24. 24.
    Mellor HR, Harris AL (2007) The role of the hypoxia-inducible BH3-only proteins BNIP3 and BNIP3L in cancer. Cancer Metastasis Rev 26:553–566. doi: 10.1007/s10555-007-9080-0 PubMedCrossRefGoogle Scholar
  25. 25.
    Nishi K, Oda T, Takabuchi S, Oda S, Fukuda K, Adachi T, Semenza GL, Shingu K, Hirota K (2008) LPS induces hypoxia-inducible factor 1 activation in macrophage-differentiated cells in a reactive oxygen species-dependent manner. Antioxid Redox Signal 10:983–995. doi: 10.1089/ars.2007.1825 PubMedCrossRefGoogle Scholar
  26. 26.
    Page EL, Robitaille GA, Pouyssegur J, Richard DE (2002) Induction of hypoxia-inducible factor-1α by transcriptional and translational mechanisms. J Biol Chem 277:48403–48409. doi: 10.1074/jbc.M209114200 PubMedCrossRefGoogle Scholar
  27. 27.
    Petronilli V, Miotto G, Canton M, Brini M, Colonna R, Bernardi P, Di LF (1999) Transient and long-lasting openings of the mitochondrial permeability transition pore can be monitored directly in intact cells by changes in mitochondrial calcein fluorescence. Biophys J 76:725–734PubMedCrossRefGoogle Scholar
  28. 28.
    Peyssonnaux C, Cejudo-Martin P, Doedens A, Zinkernagel AS, Johnson RS, Nizet V (2007) Cutting edge: essential role of hypoxia inducible factor-1α in development of lipopolysaccharide-induced sepsis. J Immunol 178:7516–7519PubMedGoogle Scholar
  29. 29.
    Riazy M, Chen JH, Steinbrecher UP (2009) VEGF secretion by macrophages is stimulated by lipid and protein components of OxLDL via PI3-kinase and PKCς activation and is independent of OxLDL uptake. Atherosclerosis 204:47–54. doi: 10.1016/j.atherosclerosis.2008.08.004 PubMedCrossRefGoogle Scholar
  30. 30.
    Ross R (1999) Atherosclerosis—an inflammatory disease. N Engl J Med 340:115–126PubMedCrossRefGoogle Scholar
  31. 31.
    Rossig L, Dimmeler S, Zeiher AM (2001) Apoptosis in the vascular wall and atherosclerosis. Basic Res Cardiol 96:11–22. doi: 10.1007/s003950170073 PubMedCrossRefGoogle Scholar
  32. 32.
    Sanson M, Ingueneau C, Vindis C, Thiers JC, Glock Y, Rousseau H, Sawa Y, Bando Y, Mallat Z, Salvayre R, Negre-Salvayre A (2008) Oxygen-regulated protein-150 prevents calcium homeostasis deregulation and apoptosis induced by oxidized LDL in vascular cells. Cell Death Differ 15:1255–1265. doi: 10.1038/cdd.2008.36 PubMedCrossRefGoogle Scholar
  33. 33.
    Shatrov VA, Sumbayev VV, Zhou J, Brune B (2003) Oxidized low-density lipoprotein (oxLDL) triggers hypoxia-inducible factor-1α (HIF-1α) accumulation via redox-dependent mechanisms. Blood 101:4847–4849. doi: 10.1182/blood-2002-09-2711 PubMedCrossRefGoogle Scholar
  34. 34.
    Sluimer JC, Gasc JM, van Wanroij JL, Kisters N, Groeneweg M, Sollewijn Gelpke MD, Cleutjens JP, van den Akker LH, Corvol P, Wouters BG, Daemen MJ, Bijnens AP (2008) Hypoxia, hypoxia-inducible transcription factor, and macrophages in human atherosclerotic plaques are correlated with intraplaque angiogenesis. JACC 51:1258–1265. doi: 10.1016/j.jacc.2007.12.025 PubMedGoogle Scholar
  35. 35.
    Takeda N, O’Dea EL, Doedens A, Kim JW, Weidemann A, Stockmann C, Asagiri M, Simon MC, Hoffmann A, Johnson RS (2010) Differential activation and antagonistic function of HIF-{alpha} isoforms in macrophages are essential for NO homeostasis. Genes Dev 24:491–501. doi: 10.1101/gad.1881410 PubMedCrossRefGoogle Scholar
  36. 36.
    Tsukamoto Y, Kuwabara K, Hirota S, Ikeda J, Stern D, Yanagi H, Matsumoto M, Ogawa S, Kitamura Y (1996) 150-kD oxygen-regulated protein is expressed in human atherosclerotic plaques and allows mononuclear phagocytes to withstand cellular stress on exposure to hypoxia and modified low density lipoprotein. J Clin Invest 98:1930–1941. doi: 10.1172/JCI118994 PubMedCrossRefGoogle Scholar
  37. 37.
    Uchida T, Rossignol F, Matthay MA, Mounier R, Couette S, Clottes E, Clerici C (2004) Prolonged hypoxia differentially regulates hypoxia-inducible factor (HIF)-1α and HIF-2α expression in lung epithelial cells: implication of natural antisense HIF-1α. J Biol Chem 279:14871–14878. doi: 10.1074/jbc.M400461200 PubMedCrossRefGoogle Scholar
  38. 38.
    Vink A, Schoneveld AH, Lamers D, Houben AJS, van der Groep P, van Diest PJ, Pasterkamp G (2007) HIF-1a expression is associated with an atheromatous inflammatory plaque phenotype and upregulated in activated macrophages. Atherosclerosis 195:e69–e75. doi: 10.1016/j.atherosclerosis.2007.05.026 PubMedCrossRefGoogle Scholar
  39. 39.
    Willert M, Augstein A, Poitz DM, Schmeisser A, Strasser RH, Braun-Dullaeus RC (2010) Transcriptional regulation of Pim-1 kinase in vascular smooth muscle cells and its role for proliferation. Basic Res Cardiol 105:267–277. doi: 10.1007/s00395-009-0055-x PubMedCrossRefGoogle Scholar
  40. 40.
    Zhang C (2008) The role of inflammatory cytokines in endothelial dysfunction. Basic Res Cardiol 103:398–406. doi: 10.1007/s00395-008-0733-0 PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • David M. Poitz
    • 1
    Email author
  • Antje Augstein
    • 1
  • Sönke Weinert
    • 1
    • 2
  • Rüdiger C. Braun-Dullaeus
    • 1
    • 2
  • Ruth H. Strasser
    • 1
  • Alexander Schmeisser
    • 1
    • 2
  1. 1.Department of Internal Medicine and CardiologyUniversity of Technology DresdenDresdenGermany
  2. 2.Internal Medicine, Department of Cardiology, Angiology and PneumologyMagdeburg UniversityMagdeburgGermany

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