Kidney and Blood Pressure Research

Original Paper

Free Access

C-Reactive Protein Induces Interleukin-6 and Thrombospondin-1 Protein and mRNA Expression through Activation of Nuclear Factor-ĸB in HK-2 Cells

Wang H.a · Chen D.a · Zhao M.a · Shu S.b · Xiong S.a · Gan X.a · Chao S.a · Cao J.a

Author affiliations

aDepartment of Cardiology, Zhongnan Hospital of Wuhan University, and bState Key Laboratory of Freshwater Ecology and Biotechnology, Wuhan, People’s Republic of China

Corresponding Author

Hai-rong Wang

Department of Cardiology

Zhongnan Hospital of Wuhan University

169 Dong Hu Road, Wuhan 430071 (People’s Republic of China)

Tel. +86 27 6781 3073, E-Mail wanghai2192@sina.com

Keywords: Mitogen-activated protein kinaseNuclear factor-kappa BInterleukin-6Thrombospondin-1C-reactive proteinRenal tubular epithelial cells

Related Articles for ""
Kidney Blood Press Res 2012;35:211–219
https://doi.org/10.1159/000332402

Abstract

Background: Although C-reactive protein (CRP) is significantly increased in patients with diabetic nephropathy, whether CRP exerts direct proinflammatory effects on human renal tubular epithelial cells (HK-2 cells) is still unclear. Methods: HK-2 cells were incubated with purified CRP at clinically relevant concentrations (0, 5, 10, 20 and 40 µg/ml). The protein and transcript levels of thrombospondin-1 (TSP-1) and interleukin-6 (IL-6) were determined by ELISA and RT-PCR. Phosphorylation of p38MAPK was investigated through Western blot analysis in HK-2 cells induced by CRP. The activation of nuclear factor-kappa B (NF-ĸB) was studied via EMSA. A specific p38MAPK inhibitor (SB203580) and an NF-ĸB inhibitor (PDTC; pyrrolidine dithiocarbamate) were used to analyze the signal transduction in CRP induction. To explore the direct or indirect role of CRP in HK-2 cells, IL-6 or TSP-1 antibodies were used. The expression of IL-6, TSP-1 and transforming growth factor-β1 (TGF-β1) were determined through Western blot analysis in HK-2 cells. Results: In HK-2 cells, purified CRP significantly induced protein release and mRNA expression of IL-6 and TSP-1 in a dose- and time-dependent manner. TGF-β1 protein was overexpressed in HK-2 cells induced by CRP, which cannot be inhibited by IL-6 or TSP-1 antibodies. CRP triggered phosphorylation of p38MAPK and activation of NF-ĸB-mediated signal transduction. SB203580 (5 µM) and PDTC (50 µM) efficiently suppressed those effects of CRP in HK-2 cells. Conclusions: CRP induces IL-6 and TSP-1 protein release and mRNA expression from HK-2 cells via activation of the p38MAPK and NF-ĸB signaling pathways and TGF-β1 was highly expressed in HK-2 cells, suggesting that CRP plays an important role in the propagation and prolongation of inflammation in renal fibrosis.

© 2012 S. Karger AG, Basel

References

  1. Lin Lu, Li Jin Pu, Xue Wei Xu, et al: Association of serum levels of glycated albumin, C-reactive protein and tumor necrosis factor-α with the severity of coronary artery disease and renal impairment in patients with type 2 diabetes mellitus. Clin Biochem 2007;40:810–816.
    External Resources
  2. Busch M, Franke S, Müller A, et al: Potential cardiovascular risk factors in chronic kidney disease: AGEs, total homocysteine and metabolites, and the C-reactive protein. Kidney Int 2004;66:338–347.
    External Resources
  3. Peppa M, Uribarri J, Cai W, Lu M, Vlassara H: Glycoxidation and inflammation in renal failure patients. Am J Kidney Dis 2004;43:690–695.
    External Resources
  4. Ridker PM, Cushman M, Stampfer MJ, Tracy RP, Hennekens CH: Plasma concentration of C-reactive protein and risk of developing peripheral vascular disease. Circulation 1998;97:425–428.
    External Resources
  5. Ridker PM, Hennekens CH, Buring JE, Rifai N: C-reactive protein and other markers of inflammation in the prediction of cardiovascular disease in women. N Engl J Med 2000;342:836–843.
    External Resources
  6. Biasucci LM, Liuzzo G, Buffon A, Maseri A: The variable role of inflammation in acute coronary syndromes and in restenosis. Semin Interv Cardiol 1999;4:105–110.
    External Resources
  7. Gussekloo J, Schaap MCL, Frölich M, Blauw GJ, Westendorp RGJ: C-reactive protein is a strong but nonspecific risk factor of fatal stroke in elderly persons. Arterioscler Thromb Vasc Biol 2000;20:1047–1051.
    External Resources
  8. Kawahara K, Biswas KK, Unoshima M, et al: C-reactive protein induces high-mobility group box-1 protein release through activation of p38MAPK in macrophage RAW264.7 cells. Cardiovasc Pathol 2008;17:129–138.
    External Resources
  9. Ridker PM, Hennekens CH, Buring JE, Rifai N: C-reactive protein increases plasminogen activator inhibitor-1 expression in human endothelial cells. Thromb Res 2008;122:125–133.
    External Resources
  10. Wang H, Li J, Huang C, et al: Fluvastatin inhibits the expression of tumor necrosis factor-alpha and activation of nuclear factor-kappa B in human endothelial cells stimulated by C-reactive protein. Clin Chim Acta 2005;353:53–60.
    External Resources
  11. Li J, Wang H, Huang C, et al: Enhanced inflammatory response of blood monocytes to C-reactive protein in patients with unstable angina. Clin Chim Acta 2005;352:127–133.
    External Resources
  12. Jinde K, Nikolic-Paterson DJ, Huang XR, et al: Tubular phenotypic change in progressive tubulointerstitial fibrosis in human glomerulonephritis. Am J Kid Dis 2001;38:761–769.
    External Resources
  13. Stenina OI, Krukovets I, Wang K, et al: Increased expression of thrombospondin-1 in vessel wall of diabetic Zucker rat. Circulation 2003;107:3209–3215.
    External Resources
  14. Janabi M, Yamashita S, Hirano K, et al: Oxidized LDL-induced NF-kappaB activation and monocyte-derived macrophages from CD36-deficient patients. Arterioscler Thromb Vasc Biol 2000;20:1953–1960.
    External Resources
  15. Han KH, Hong K-H, Park J-H, et al: C-reactive protein promotes monocyte chemoattractant protein-1-mediated chemotaxis through upregulating CC chemokine receptor 2 expression in human monocytes. Circulation 2004;109:2566–2571.
    External Resources
  16. Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(–Delta Delta C(T)) method. Methods 2001;25:402–408.
    External Resources
  17. Inoue K, Kawahara K, Biswas KK, Ando K, Mitsudo K, Nobuyoshi M, Maruyama I: HMGB1 expression by activated vascular smooth muscle cells in advanced human atherosclerosis plaques. Cardiovasc Pathol 2007;16:136–143.
    External Resources
  18. Yudkin JS, Kumari M, Humphries SE, Mohamed-Ali V: Inflammation, obesity, stress and coronary heart disease: is interleukin-6 the link? Atherosclerosis 2000;148:209–214.
    External Resources
  19. Beg AA, Finco TS, Nantermet PV, Baldwin AS Jr: Tumor necrosis factor and interleukin-1 lead to phosphorylation and loss of ĸBα: a mechanism for NF-ĸB activation. Mol Cell Biol 1993;13:3301–3310.
    External Resources
  20. Bang R, Marnell L, Mold C, et al: Analysis of binding sites in human C-reactive protein for Fc{gamma}RI, Fc{gamma}RIIA, and C1q by site-directed mutagenesis. J Biol Chem 2005;280:25095–25102.
    External Resources
  21. Dalla Vestra M, Fioretto P: Diabetic nephropathy: renal structural studies in type 1 and type 2 diabetic patients. Int Congr Ser 2005;1253:163–169.
    External Resources
  22. Fioretto P, Bruseghin M, Barzon I, Arboita M, Dalla Vestraa M: Diabetic nephropathy: an update on renal structure. Int Congr Ser 2007;1303:51–59.
    External Resources
  23. Bonnefoy A, Moura R, Hoylaerts MF: The evolving role of thrombospondin-1 in hemostasis and vascular biology. Cell Mol Life Sci 2008,65:713–727.
    External Resources
  24. Vickers MA, Green FR, Terry C, et al: Genotype at a promoter polymorphism of the interleukin-6 gene is associated with baseline levels of plasma C-reactive protein. Cardiovasc Res 2002;53:1029–1034.
    External Resources
  25. Yeh ET, Anderson HV, Pasceri V, Willerson JT: C-reactive protein: linking inflammation to cardiovascular complications. Circulation 2001;104:974–975.
    External Resources
  26. Tomai F, Crea F, Gaspardone A, et al: Unstable angina and elevated C-reactive protein levels predict enhanced vasoreactivity of the culprit lesion. Circulation 2002;104:1471–1476.
    External Resources
  27. Nishikawa M, Myoui A, Tomita T, et al: Prevention of the onset and progression of collagen-induced arthritis in rats by the potent p38 mitogen-activated protein kinase inhibitor FR167653. Arthritis Rheum 2003;48:2670–2681.
    External Resources
  28. Kuldo JM, Westra J, Asgeirsdottir SA, et al: Differential effects of NF-{kappa}B and p38 MAPK inhibitors and combinations thereof on TNF-{alpha}- and IL-1{beta}-induced proinflammatory status of endothelial cells in vitro. Am J Physiol Cell Physiol 2005;289:1229–1239.
    External Resources
  29. Pepys MB, Hirschfield GM, Tennent GA, et al: Targeting C-reactive protein for the treatment of cardiovascular disease. Nature 2006;440:1217–1221.
    External Resources
  30. Johansen C, Funding AT, Otkjaer K, et al: Protein expression of TNF-{alpha} in psoriatic skin is regulated at a posttranscriptional level by MAPK-activated protein kinase 2. J Immunol 2006;176:1431–1438.
    External Resources
  31. Perlstein RS, Whitnall MH, Abrams JS, Mougey EH, Neta R: Synergistic roles of interleukin-6, interleukin-1, and tumor necrosis factor in the adrenocorticotropin response to bacterial lipopolysaccharide in vivo: synergistic roles of interleukin-6, interleukin-1, and tumor necrosis factor in the adrenocorticotropin response to bacterial lipopolysaccharide in vivo. Endocrinology 1993;132:946–952.
    External Resources
  32. Münzel D, Lehle K, Haubner F, et al: Impact of diabetic serum on endothelial cells: An in-vitro-analysis of endothelial dysfunction in diabetes mellitus type 2. Biochem Biophys Res Commun 2007;362:238–244.
    External Resources
  33. Brennan P, O’Neill LA: 2-mercaptoethanol restores the ability of nuclear factor kappa B (NF kappa B) to bind DNA in nuclear extracts from interleukin 1-treated cells incubated with pyrollidine dithiocarbamate (PDTC): evidence for oxidation of glutathione in the mechanism of inhibition of NF kappa B by PDTC. Biochem J 1996;320:975–981.
    External Resources
  34. Kalinina N, Agrotis A, Antropova Y, DiVitto G, Kanellakis P, Kostolias G, et al: Increased expression of the DNA-binding cytokine IL-6 in human atherosclerotic lesions. Arterioscler Thromb Vasc Biol 2004;24:241–247.
    External Resources
  35. Verma S, Li SH, Badiwala MV, et al: Endothelin antagonism and interleukin-6 inhibition attenuate the proatherogenic effects of C-reactive protein. Circulation 2002;23:1890–1896.
    External Resources
  36. Thieringer R, Fenyk-Melody JE, Le Grand CB, et al: Activation of peroxisome proliferator-activated receptor gamma does not inhibit IL-6 or TNF-alpha responses of macrophages to lipopolysaccharide in vitro or in vivo. Immunology 2000;164:1046–1054.
    External Resources
  37. Tang D, Shi Y, Kang R, et al: Hydrogen peroxide stimulates macrophages and monocytes to actively release HMGB. J Leukoc Biol 2007;81:741–747.
    External Resources
  38. Argania H, Aghaeishahsavarib M, Noroozianavval M, et al: Effects of losartan and enalapril on high-sensitivity C-reactive protein and total antioxidant in renal transplant recipients with renin-angiotensin system polymorphisms. Transplant Proc 2008;40:16–21.
    External Resources
  39. Maeda S: Do inflammatory cytokine genes confer susceptibility to diabetic nephropathy? Kidney Int 2008;74:413–415.
    External Resources
  40. Mineo C, Gormley AK, Yuhanna IS, et al: FcgammaRIIB mediates C-reactive protein inhibition of endothelial NO synthase. Circ Res 2005;97:1124–1131.
    External Resources
  41. Baer PC, Gauer S, Wegner B, Schubert R, Geiger H: C-reactive protein induced activation of MAP-K and RANTES in human renal distal tubular epithelial cells in vitro. Clin Nephrol 2006;66:177–183.
    External Resources
  42. Chang JW, Kim CS, Kim SB, et al: C-reactive protein induces NF-kappaB activation through intracellular calcium and ROS in human mesangial cells. Nephron Exp Nephrol 2005;101:e165–e172.
    External Resources
  43. Patel DN, King CA, Bailey SR, et al: Interleukin-17 stimulates C-reactive protein expression in hepatocytes and smooth muscle cells via p38 MAPK and ERK1/2-dependent NF-ĸB and C/EBPβ. J Biol Chem 2007;282:27229–27238.
    External Resources

Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: January 24, 2011
Accepted: August 28, 2011
Published online: January 03, 2012
Issue release date: May 2012

Number of Print Pages: 9
Number of Figures: 5
Number of Tables: 1

ISSN: 1420-4096 (Print)
eISSN: 1423-0143 (Online)

For additional information: https://www.karger.com/KBR

Copyright / Drug Dosage / Disclaimer

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Article Tools
Article Details

2012, Vol.35, No. 4

May 2012

Article

TAP Icon This Article is Part of
Recommend This
Open Access Gateway
TOP