Skip to main content
Log in

Secretion of MCP-1 and IL-6 by cytokine stimulated production of reactive oxygen species in endothelial cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Endothelial cells are known to produce reactive oxygen species by several mechanisms. Functional consequences of increased production of reactive oxygen species were investigated in vitro after stimulation with several proinflammatory cytokines. Time dependent increases in DCF-fluorescence as a measure of reactive oxygen load were quantified in single cells after incubation with TNF-α, IL-1 and IFN-γ. The increased DCF-fluorescence was inhibited by cell permeant antioxidative substances Tiron and Tempol. NMMA, an inhibitor of nitric oxide synthase reduced endothelial DCF-fluorescence only marginally, indicating a minor participation of nitric oxide production in this detection system. Cytokine induced endothelial DCF-fluorescence increased in the presence of NADH, whereas coincubation with NADPH or xanthine was without effect. Flavoenzyme inhibitor diphenyliodonium abolished stimulated DCF-fluorescence. Cytokine induced release of MCP-1 and IL-6 by endothelial cells was completely inhibited in the presence of Tiron and Tempol, whereas NMMA was less effective. Collectively these data indicate that cytokine stimulated endothelial cells increase their reactive oxygen species production probably via NADH oxidase and this production may critically be involved in the secretion of MCP-1 and IL-6.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Rosen GM, Freeman BA: Detection of superoxide generated by endothelial cells. Proc Natl Acad Sci USA 81: 7269-7273, 1984

    Google Scholar 

  2. Panus PC, Radi R, Chumley PH, Lillard RH, Freeman BA: Detection of H2O2release from vascular endothelial cells. Free Rad Biol Med 14: 217-223, 1993

    Google Scholar 

  3. Arroyo CM, Carmichael AJ, Bouscarel B, Liang JH, Weglicki WB: Endothelial cells as a source of oxygen-free radicals. An ESR study. Free Rad Res Commun 9: 287-296, 1990

    Google Scholar 

  4. Kloner RA, Przyklenk K, Whittaker P: Deleterious effects of oxygen radicals in ischemia/reperfusion. Resolved and unresolved issues. Circulation 80: 1115-1127, 1989

    Google Scholar 

  5. Wang JH, Redmond HP, Watson RW, Bouchier Hayes D: Induction of human endothelial cell apoptosis requires both heat shock and oxidative stress responses. Am J Physiol 272: C1543-C1551, 1997

    Google Scholar 

  6. Beckman-JS; Beckman-TW; Chen-J; Marshall-PA; Freeman-BA: Apparent hydroxyl radical production by peroxynitrite: Implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci USA 87: 1620-1624, 1990

    Google Scholar 

  7. Halliwell B: Free radicals, antioxidants, and human disease: Curiosity, cause, or consequence? Lancet 344: 721-724, 1994

    Google Scholar 

  8. Bone RC: Toward a theory regarding the pathogenesis of the systemic inflammatory response syndrome: What we do and do not know about cytokine regulation. Crit Care Med 24: 163-172, 1996

    Google Scholar 

  9. Matsubara T, Ziff M: Increased superoxide anion release from human endothelial cells in response to cytokines. J Immunol 137: 3295-3298, 1986

    Google Scholar 

  10. Murphy HS, Shayman JA, Till GO, Mahrougui M, Owens CB, Ryan US, Ward PA: Superoxide responses of endothelial cells to C5a and TNF-alpha: Divergent signal transduction pathways. Am J Physiol 263: L51-L59, 1992

    Google Scholar 

  11. Volk T, Hensel M, Kox WJ: Transient Ca2+ changes in endothelial cells induced by low doses of reactive oxygen species: Role of hydrogen peroxide. Mol Cell Biochem 171: 11-21, 1997

    Google Scholar 

  12. Edgell CJ, McDonald CC, Graham JB: Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc Natl Acad Sci USA 80: 3734-3737, 1983

    Google Scholar 

  13. Cines DB, Pollak ES, Buck CA, Loscalzo J, Zimmerman GA, McEver RP, Pober JS, Wick TM, Konkle BA, Schwartz BS, Barnathan ES, McCrae KR, Hug BA, Schmidt AM, Stern DM: Endothelial cells in physiology and in the pathophysiology of vascular disorders. Blood 91: 3527-3561, 1998

    Google Scholar 

  14. Bossink AW, Paemen L, Jansen PM, Hack CE, Thijs LG, Van Damme J: Plasma levels of the chemokines monocyte chemotactic proteins-1 and-2 are elevated in human sepsis. Blood 86: 3841-3847, 1995

    Google Scholar 

  15. Goodman RB, Strieter RM, Martin DP, Steinberg KP, Milberg JA, Maunder RJ, Kunkel SL, Walz A, Hudson LD, Martin TR: Inflammatory cytokines in patients with persistence of the acute respiratory distress syndrome. Am J Resp Crit Care Med 154: 602-611, 1996

    Google Scholar 

  16. Goode HF, Webster NR: Free radicals and antioxidants in sepsis. Crit Care Med 21: 1770-1776, 1993

    Google Scholar 

  17. Taylor DE, Piantadosi CA: Oxidative metabolism in sepsis and sepsis syndrome. J Crit Care 10: 122-135, 1995

    Google Scholar 

  18. Cosentino F, Hishikawa K, Katusic ZS, Luscher TF: High glucose increases nitric oxide synthase expression and superoxide anion generation in human aortic endothelial cells. Circulation 96: 25-28, 1997

    Google Scholar 

  19. Rajagopalan S, Kurz S, Munzel T, Tarpey M, Freeman BA, Griendling KK, Harrison DG: Angiotensin II-mediated hypertension in the rat increases vascular superoxide production via membrane NADH/NADPH oxidase activation. Contribution to alterations of vasomotor tone. J Clin Invest 97: 1916-1923, 1996

    Google Scholar 

  20. Panus PC, Radi R, Chumley PH, Lillard RH, Freeman BA: Detection of H2O2 release from vascular endothelial cells. Free Rad Biol Med 14: 217-223, 1993

    Google Scholar 

  21. Royall JA, Gwin PD, Parks DA, Freeman-BA: Responses of vascular endothelial oxidant metabolism to lipopolysaccharide and tumor necrosis factor-alpha. Arch Biochem Biophys 294: 686-694, 1992

    Google Scholar 

  22. Arai T, Kelly SA, Brengman ML, Takano M, Smith EH, Goldschmidt Clermont PJ, Bulkley GB: Ambient but not incremental oxidant generation effects intercellular adhesion molecule 1 induction by tumour necrosis factor alpha in endothelium. Biochem J 331: 853-861, 1998

    Google Scholar 

  23. Galley HF, Walker BE, Howdle PD, Webster NR: Regulation of nitric oxide synthase activity in cultured human endothelial cells: Effect of antioxidants. Free Rad Biol Med 21: 97-101, 1996

    Google Scholar 

  24. Ito A, Tsao PS, Adimoolam S, Kimoto M, Ogawa T, Cooke JP: Novel mechanism for endothelial dysfunction: Dysregulation of dimethylarginine dimethylaminohydrolase. Circulation 99: 3092-3095, 1999

    Google Scholar 

  25. Anema SM, Walker SW, Howie AF, Arthur JR, Nicol F, Beckett GJ: Thioredoxin reductase is the major selenoprotein expressed in human umbilical-vein endothelial cells and is regulated by protein kinase C. Biochem J 342: 111-117, 1999

    Google Scholar 

  26. Frigerio S, Gelati M, Ciusani E, Corsini E, Dufour A, Massa G, Salmaggi A: Immunocompetence of human microvascular brain endothelial cells: Cytokine regulation of IL-1beta, MCP-1, IL-10, sICAM-1 and sVCAM-1. J Neurol245: 727-730, 1998

    Google Scholar 

  27. Krishnaswamy G, Smith JK, Mukkamala R, Hall K, Joyner W L Y, Chi DS: Multifunctional cytokine expression by human coronary endothelium and regulation by monokines and glucocorticoids. Microvasc Res 55: 189-200, 1998

    Google Scholar 

  28. Sanders SP, Zweier JL, Kuppusamy P, Harrison SJ, Bassett DJ, Gabrielson EW, Sylvester JT: Hyperoxic sheep pulmonary microvascular endothelial cells generate free radicals via mitochondrial electron transport. J Clin Invest 91: 46-52, 1993

    Google Scholar 

  29. Zweier JL, Kuppusamy P, Thompson-Gorman S, Klunk D, Lutty GA: Measurement and characterization of free radical generation in reoxygenated human endothelial cells. Am J Physiol 266: C700-C708, 1994

    Google Scholar 

  30. Xia Y, Tsai AL, Berka V, Zweier JL: Superoxide generation from endothelial nitric-oxide synthase. A Ca2+/calmodulin-dependent and tetrahydrobiopterin regulatory process. J Biol Chem 273: 25804-25808, 1998

    Google Scholar 

  31. Mohazzab KM, Kaminski PM, Wolin MS: NADH oxidoreductase is a major source of superoxide anion in bovine coronary artery endothelium. Am J Physiol 266: H2568-H2572, 1994

    Google Scholar 

  32. Darley-Usmar V, Wiseman H, Halliwell B: Nitric oxide and oxygen radicals: A question of balance. FEBS Lett 369: 131-135, 1995

    Google Scholar 

  33. Cheng JJ, Wung BS, Chao YJ, Wang DL: Cyclic strain-induced reactive oxygen species involved in ICAM-1 gene induction in endothelial cells. Hypertension 31: 125-130, 1998

    Google Scholar 

  34. Weber C, Erl W, Pietsch A, Strobel M, Ziegler-Heitbrock HW, Weber PC: Antioxidants inhibit monocyte adhesion by suppressing nuclear factor-kappa B mobilization and induction of vascular cell adhesion molecule-1 in endothelial cells stimulated to generate radicals. Arterioscler Thromb 14: 1665-1673, 1994

    Google Scholar 

  35. Flohe L, Brigelius-Flohe R, Saliou C, Traber MG, Packer L L: Redox regulation of NF-kappa B activation. Free Rad Biol Med 22: 1115-1126, 1997

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Volk, T., Hensel, M., Schuster, H. et al. Secretion of MCP-1 and IL-6 by cytokine stimulated production of reactive oxygen species in endothelial cells. Mol Cell Biochem 206, 105–112 (2000). https://doi.org/10.1023/A:1007059616914

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1007059616914

Navigation