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Role of NADPH oxidase in interleukin-4-induced monocyte chemoattractant protein-1 expression in vascular endothelium

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Abstract

Objective and design

The pro-oxidative and pro-inflammatory pathways in vascular endothelium have been implicated in the development of atherosclerosis. In the present study, we investigated effect of interleukin-4 (IL-4) on monocyte chemoattractant protein-1 (MCP-1) expression in vascular endothelium and examined the role of distinct sources of reactive oxygen species (ROS) in this process.

Methods and results

Real-time reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay showed that IL-4 significantly up-regulated mRNA and protein expression of MCP-1 in human aortic endothelial cells (HAEC) and C57BL/6 mice. A significant and dose-dependent inhibition of IL-4-induced MCP-1 expression was observed in HAEC pre-treated with antioxidants, such as pyrrolidine dithiocarbamate and epigallocatechin gallate, indicating that IL-4-induced MCP-1 expression is mediated via a ROS-dependent mechanism. Additionally, pharmacological inhibitors of NADPH oxidase (NOX) significantly attenuated IL-4-induced MCP-1 expression in HAEC. Furthermore, the disruption of the NOX gene dramatically reduced IL-4-induced MCP-1 expression in NOX knockout mice (B6.129S6-Cybbtm1Din/J). In contrast, overexpression of MCP-1 in IL-4-stimulated HAEC was not affected by inhibiting other ROS generating pathways, such as xanthine oxidase and the mitochondrial electron transport chain.

Conclusions

These results demonstrate that IL-4 up-regulates MCP-1 expression in vascular endothelium through NOX-mediated ROS generation.

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References

  1. Lee YW, Hennig B, Toborek M. Redox-regulated mechanisms of interleukin-4-induced MCP-1 expression in human vascular endothelial cells. Am J Physiol Heart Circ Physiol. 2003;284:H185–92.

    CAS  PubMed  Google Scholar 

  2. Rollins BJ, Pober JS. Interleukin-4 induces the synthesis and secretion of MCP-1/JE by human endothelial cells. Am J Pathol. 1991;138:1315–9.

    CAS  PubMed  Google Scholar 

  3. Bouloumie A, Marumo T, Lafontan M, Busse R. Leptin induces oxidative stress in human endothelial cells. FASEB J. 1999;13:1231–8.

    CAS  PubMed  Google Scholar 

  4. Strieter RM, Wiggins R, Phan SH, Wharram BL, Showell HJ, Remick DG, et al. Monocyte chemotactic protein gene expression by cytokine-treated human fibroblasts and endothelial cells. Biochem Biophys Res Commun. 1989;162:694–700.

    Article  CAS  PubMed  Google Scholar 

  5. Taubman MB, Rollins BJ, Poon M, Marmur J, Green RS, Berk BC, et al. JE mRNA accumulates rapidly in aortic injury and in platelet-derived growth factor-stimulated vascular smooth muscle cells. Circ Res. 1992;70:314–25.

    CAS  PubMed  Google Scholar 

  6. Wung BS, Cheng JJ, Hsieh HJ, Shyy YJ, Wang DL. Cyclic strain-induced monocyte chemotactic protein-1 gene expression in endothelial cells involves reactive oxygen species activation of activator protein 1. Circ Res. 1997;81:1–7.

    CAS  PubMed  Google Scholar 

  7. Nelken NA, Coughlin SR, Gordon D, Wilcox JN. Monocyte chemoattractant protein-1 in human atheromatous plaques. J Clin Invest. 1991;88:1121–7.

    Article  CAS  PubMed  Google Scholar 

  8. Yla-Herttuala S, Lipton BA, Rosenfeld ME, Sarkioja T, Yoshimura T, Leonard EJ, et al. Expression of monocyte chemoattractant protein 1 in macrophage-rich areas of human and rabbit atherosclerotic lesions. Proc Natl Acad Sci USA. 1991;88:5252–6.

    Article  CAS  PubMed  Google Scholar 

  9. Takeya M, Yoshimura T, Leonard EJ, Takahashi K. Detection of monocyte chemoattractant protein-1 in human atherosclerotic lesions by an anti-monocyte chemoattractant protein-1 monoclonal antibody. Hum Pathol. 1993;24:534–9.

    Article  CAS  PubMed  Google Scholar 

  10. Gu L, Okada Y, Clinton SK, Gerard C, Sukhova GK, Libby P, et al. Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. Mol Cell. 1998;2:275–81.

    Article  CAS  PubMed  Google Scholar 

  11. Gosling J, Slaymaker S, Gu L, Tseng S, Zlot CH, Young SG, et al. MCP-1 deficiency reduces susceptibility to atherosclerosis in mice that overexpress human apolipoprotein B. J Clin Invest. 1999;103:773–8.

    Article  CAS  PubMed  Google Scholar 

  12. Boring L, Gosling J, Cleary M, Charo IF. Decreased lesion formation in CCR2−/− mice reveals a role for chemokines in the initiation of atherosclerosis. Nature. 1998;394:894–7.

    Article  CAS  PubMed  Google Scholar 

  13. Aiello RJ, Bourassa PK, Lindsey S, Weng W, Natoli E, Rollins BJ, et al. Monocyte chemoattractant protein-1 accelerates atherosclerosis in apolipoprotein E-deficient mice. Arterioscler Thromb Vasc Biol. 1999;19:1518–25.

    CAS  PubMed  Google Scholar 

  14. Martinovic I, Abegunewardene N, Seul M, Vosseler M, Horstick G, Buerke M, et al. Elevated monocyte chemoattractant protein-1 serum levels in patients at risk for coronary artery disease. Circ J. 2005;69:1484–9.

    Article  CAS  PubMed  Google Scholar 

  15. Ni W, Egashira K, Kitamoto S, Kataoka C, Koyanagi M, Inoue S, et al. New anti-monocyte chemoattractant protein-1 gene therapy attenuates atherosclerosis in apolipoprotein E-knockout mice. Circulation. 2001;103:2096–101.

    CAS  PubMed  Google Scholar 

  16. Inoue S, Egashira K, Ni W, Kitamoto S, Usui M, Otani K, et al. Anti-monocyte chemoattractant protein-1 gene therapy limits progression and destabilization of established atherosclerosis in apolipoprotein E-knockout mice. Circulation. 2002;106:2700–6.

    Article  CAS  PubMed  Google Scholar 

  17. Tedgui A, Mallat Z. Cytokines in atherosclerosis: pathogenic and regulatory pathways. Physiol Rev. 2006;86:515–81.

    Article  CAS  PubMed  Google Scholar 

  18. Buono C, Come CE, Stavrakis G, Maguire GF, Connelly PW, Lichtman AH. Influence of interferon-γ on the extent and phenotype of diet-induced atherosclerosis in the LDLR-deficient mouse. Arterioscler Thromb Vasc Biol. 2003;23:454–60.

    Article  CAS  PubMed  Google Scholar 

  19. Barks JL, McQuillan JJ, Iademarco F. TNF-α and IL-4 synergistically increase vascular cell adhesion molecule-1 expression in cultured vascular smooth muscle cells. J Immunol. 1997;159:4532–87.

    CAS  PubMed  Google Scholar 

  20. Blease K, Seybold J, Adcock IM, Hellewell PG, Burke-Gaffney A. Interleukin-4 and lipopolysaccharide synergize to induce vascular cell adhesion molecule-1 expression in human lung microvascular endothelial cells. Am J Respir Cell Mol Biol. 1998;18:620–30.

    CAS  PubMed  Google Scholar 

  21. Davenport P, Tipping PG. The role of interleukin-4 and interleukin-12 in the progression of atherosclerosis in apolipoprotein E-deficient mice. Am J Pathol. 2003;163:1117–25.

    CAS  PubMed  Google Scholar 

  22. Galéa P, Thibault G, Lacord M, Bardos P, Lebranchu Y. IL-4, but not tumor necrosis factor-alpha, increases endothelial cell adhesiveness for lymphocytes by activating a cAMP-dependent pathway. J Immunol. 1993;151:588–96.

    PubMed  Google Scholar 

  23. Hong HY, Lee HY, Kwak W, Yoo J, Na MH, So IS, et al. Phage display selection of peptides that home to atherosclerotic plaques: IL-4 receptor as a candidate target in atherosclerosis. J Cell Moll Med. 2008;12:2003–14.

    Article  CAS  Google Scholar 

  24. Huang H, Lavoie-Lamoureux A, Moran K, Lavoie JP. IL-4 stimulates the expression of CXCL-8, E-selectin, VEGF, and inducible nitric oxide synthase mRNA by equine pulmonary artery endothelial cells. Am J Physiol Lung Cell Mol Physiol. 2007;292:L1147–54.

    Article  CAS  PubMed  Google Scholar 

  25. King VL, Szilvassy SJ, Daugherty A. Interleukin-4 deficiency decreases atherosclerotic lesion formation in a site-specific manner in female LDL receptor−/− mice. Arterioscler Thromb Vasc Biol. 2002;22:456–61.

    Article  CAS  PubMed  Google Scholar 

  26. Lee YW, Eum SY, Chen KC, Hennig B, Toborek M. Gene expression profile in interleukin-4-stimulated human vascular endothelial cells. Mol Med. 2004;10:19–27.

    Article  CAS  PubMed  Google Scholar 

  27. Lee YW, Hirani AA. Role of interleukin-4 in atherosclerosis. Arch Pharm Res. 2006;29:1–15.

    Article  CAS  PubMed  Google Scholar 

  28. Lee YW, Kühn H, Hennig B, Neish AS, Toborek M. IL-4-induced oxidative stress upregulates VCAM-1 gene expression in human endothelial cells. J Mol Cell Cardiol. 2001;33:83–94.

    Article  CAS  PubMed  Google Scholar 

  29. Lee YW, Kühn H, Kaiser S, Hennig B, Daughterty A, Toborek M. Interleukin 4 induces transcription of the 15-lipoxygenase I gene in human endothelial cells. J Lipid Res. 2001;42:783–91.

    CAS  PubMed  Google Scholar 

  30. Masinovsky B, Urdal D, Gallatin WM. IL-4 acts synergistically with IL-1β to promote lymphocyte adhesion to microvascular endothelium by induction of vascular cell adhesion molecule-1. J Immunol. 1990;145:2886–95.

    CAS  PubMed  Google Scholar 

  31. Paul WE. Interleukin-4: a prototypic immunoregulatory lymphokine. Blood. 1991;77:1859–70.

    CAS  PubMed  Google Scholar 

  32. Rey FE, Cifuentes ME, Kiarash A, Quinn MT, Pagano PJ. Novel competitive inhibitor of NAD(P)H oxidase assembly attenuates vascular O2 − and systolic blood pressure in mice. Circ Res. 2001;89:408–11.

    Article  CAS  PubMed  Google Scholar 

  33. Rocken M, Racke M, Shevach EM. IL-4-induced immune deviation as antigen-specific therapy for inflammatory autoimmune disease. Immunol Today. 1996;17:225–31.

    Article  CAS  PubMed  Google Scholar 

  34. Walch L, Massade L, Dufilho M, Brunet A, Rendu F. Pro-atherogenic effect of interleukin-4 in endothelial cells: modulation of oxidative stress, nitric oxide and monocyte chemoattractant protein-1 expression. Atherosclerosis. 2006;187:285–91.

    Article  CAS  PubMed  Google Scholar 

  35. Lee YW, Lee WH, Kim PH. Oxidative mechanisms of IL-4-induced IL-6 expression in vascular endothelium. Cytokine. 2010;49:73–9.

    Article  CAS  PubMed  Google Scholar 

  36. Lee YW, Kühn H, Hennig B, Daughterty A, Toborek M. IL-4 induces apoptosis of endothelial cells through the caspase-3-dependent pathway. FEBS Lett. 2000;485:122–6.

    Article  CAS  PubMed  Google Scholar 

  37. Sasaguri T, Arima N, Tanimoto A, Shimajiri S, Hamada T, Sasaguri Y. A role for interleukin 4 in production of matrix metalloproteinase 1 by human aortic smooth muscle cells. Atherosclerosis. 1998;138:247–53.

    Article  CAS  PubMed  Google Scholar 

  38. Chen CC, Manning AM. TGF-β1, IL-10 and IL-4 differentially modulate the cytokine-induced expression of IL-6 and IL-8 in human endothelial cells. Cytokine. 1996;8:58–65.

    Article  CAS  PubMed  Google Scholar 

  39. Bennett BL, Cruz R, Lacson RG, Manning AM. Interleukin-4 suppression of tumor necrosis factor α-stimulated E-selectin gene transcription is mediated by STAT6 antagonism of NF-κB. J Biol Chem. 1997;272:10212–9.

    Article  CAS  PubMed  Google Scholar 

  40. Wright PS, Cooper JR, Kropp KE, Busch SJ. Induction of vascular cell adhesion molecule-1 expression by IL-4 in human aortic endothelial cells is not associated with increased nuclear NF-κB levels. J Cell Physiol. 1999;180:381–9.

    Article  CAS  PubMed  Google Scholar 

  41. Toborek M, Lee YW, Kaiser S, Hennig B. Measurement of inflammatory properties of fatty acids in human endothelial cells. Methods Enzymol. 2002;352:198–219.

    Article  CAS  PubMed  Google Scholar 

  42. Deng X, Li H, Tang YW. Cytokine expression in respiratory syncytial virus-infected mice as measured by quantitative reverse-transcriptase PCR. J Virol Methods. 2003;107:141–6.

    Article  CAS  PubMed  Google Scholar 

  43. Lee YW, Lee WH. Protective effects of genistein on pro-inflammatory pathways in human brain microvascular endothelial cells. J Nutr Biochem. 2008;19:819–25.

    Article  CAS  PubMed  Google Scholar 

  44. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆CT method. Methods. 2001;25:402–8.

    Article  CAS  PubMed  Google Scholar 

  45. Rollins BJ. Chemokines. Blood. 1997;90:909–28.

    CAS  PubMed  Google Scholar 

  46. Gu L, Tseng SC, Rollins BJ. Monocyte chemoattractant protein-1. Chem Immunol. 1999;72:7–29.

    Article  CAS  PubMed  Google Scholar 

  47. Bursill CA, Channon KM, Greaves DR. The role of chemokines in atherosclerosis: recent evidence from experimental models and population genetics. Curr Opin Lipidol. 2004;15:145–9.

    Article  CAS  PubMed  Google Scholar 

  48. Sheikine YA, Hansson GK. Chemokines as potential therapeutic targets in atherosclerosis. Curr Drug Targets. 2006;7:13–27.

    Article  CAS  PubMed  Google Scholar 

  49. Braunersreuther V, Mach F, Steffens S. The specific role of chemokines in atherosclerosis. Thromb Haemost. 2007;97:714–21.

    CAS  PubMed  Google Scholar 

  50. Gimbrone MA, Bevilacqua MP, Cybulsky MI. Endothelial-dependent mechanisms of leukocyte adhesion in inflammation and atherosclerosis. Ann NY Acad Sci. 1990;598:77–85.

    Article  PubMed  Google Scholar 

  51. Iseki A, Kambe F, Okumura K, Niwata S, Yamamoto R, Hayakawa T, et al. Pyrrolidine dithiocarbamate inhibits TNF-α-dependent activation of NF-κB by increasing intracellular copper level in human aortic smooth muscle cells. Biochem Biophys Res Commun. 2000;276:88–92.

    Article  CAS  PubMed  Google Scholar 

  52. Tipoe GL, Leung TM, Hung MW, Fung ML. Green tea polyphenols as an anti-oxidant and anti-inflammatory agent for cardiovascular protection. Cardiovasc Hematol Disord Drug Targets. 2007;7:135–44.

    CAS  PubMed  Google Scholar 

  53. Basta G, Lazzerini G, Del Turco S, Ratto GM, Schmidt AM, De Caterina R. At least 2 distinct pathways generating reactive oxygen species mediate vascular cell adhesion molecule-1 induction by advanced glycation end products. Arterioscler Thromb Vasc Biol. 2005;25:1401–7.

    Article  CAS  PubMed  Google Scholar 

  54. Hennig B, Toborek M, McClain CJ, Diana JN. Nutritional implications in vascular endothelial cell metabolism. J Am Coll Nutr. 1996;15:345–58.

    CAS  PubMed  Google Scholar 

  55. Sorescu D, Weiss D, Lassegue B, Clempus RE, Szocs K, Sorescu GP, et al. Superoxide production and expression of Nox family proteins in human atherosclerosis. Circulation. 2002;105:1429–35.

    Article  CAS  PubMed  Google Scholar 

  56. Guzik TJ, Sadowski J, Guzik B, Jopek A, Kapelak B, Przybylowski P, et al. Coronary artery superoxide production and nox isoform expression in human coronary artery disease. Arterioscler Thromb Vasc Biol. 2006;26:333–9.

    Article  CAS  PubMed  Google Scholar 

  57. Hennig B, Chow CK. Lipid peroxidation and endothelial cells injury: implication in atherosclerosis. Free Radical Biol Med. 1988;4:99–106.

    Article  CAS  Google Scholar 

  58. Thomas SR, Witting PK, Drummond GR. Redox control of endothelial function and dysfunction: molecular mechanisms and therapeutic opportunities. Antioxid Redox Signal. 2008;10:1713–65.

    Article  CAS  PubMed  Google Scholar 

  59. Heitzer T, Schlinzig T, Krohn K, Meinertz T, Munzel T. Endothelial dysfunction, oxidative stress, and risk of cardiovascular events in patients with coronary artery disease. Circulation. 2001;104:2673–8.

    Article  CAS  PubMed  Google Scholar 

  60. Yla-Herttuala S. Gene expression in atherosclerotic lesions. Hertz. 1992;17:270–6.

    CAS  Google Scholar 

  61. Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature. 1993;362:801–9.

    Article  CAS  PubMed  Google Scholar 

  62. Brinckmann R, Topp MS, Zalan I, Heydeck D, Ludwig P, Kühn H, et al. Regulation of 15-lipoxygenase expression in lung epithelial cells by interleukin-4. Biochem J. 1996;318:305–12.

    CAS  PubMed  Google Scholar 

  63. Bedard K, Krause K. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiol Rev. 2007;87:245–313.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported in part by Grants from National Institutes of Health/National Heart, Lung, and Blood Institute (HL085229) and National Science Foundation Macromolecular Interfaces with Life Sciences-Integrative Graduate Education and Research Traineeship (MILES-IGERT).

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Authors and Affiliations

  1. Laboratory of Vascular Biology, Department of Biomedical Sciences and Pathobiology, School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University (Virginia Tech), ICTAS Building (MC-0298), Stanger Street, Blacksburg, VA, 24061, USA

    Yong Woo Lee & Paul H. Kim

  2. School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA, 24061, USA

    Yong Woo Lee & Won Hee Lee

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  1. Yong Woo Lee
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Correspondence to Yong Woo Lee.

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Responsible Editor: I. Ahnfelt-Rønne.

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Lee, Y.W., Lee, W.H. & Kim, P.H. Role of NADPH oxidase in interleukin-4-induced monocyte chemoattractant protein-1 expression in vascular endothelium. Inflamm. Res. 59, 755–765 (2010). https://doi.org/10.1007/s00011-010-0187-3

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  • DOI: https://doi.org/10.1007/s00011-010-0187-3

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