Background: Glomerular mononuclear cell infiltration is associated with the development of a diffuse glomerulosclerosis in patients with diabetic nephropathy. Monocyte chemoattractant protein-1 (MCP-1) plays an important role in the recruitment and accumulation of monocytes and lymphocytes within the glomerulus. In the present study, we examined whether the ambient glucose concentration alters the expression of MCP-1 and its receptor CCR2 in primary human mesangial cells (HMC). Methods: MCP-1 mRNA expression was assessed by Northern blot and CCR2 mRNA expression by RT-PCR analysis. MCP-1 protein production was determined by ELISA. Migration studies were performed to assess functional MCP-1 receptor expression. Results: Exposure of HMC to 30 mM D-glucose led to a 30% increase in MCP-1 mRNA expression as compared to 5 mM D-glucose and osmotic controls while there was no difference in MCP-1 protein production. Simultaneously, CCR2 mRNA expression was down-regulated in HMC exposed to 30 mM D-glucose. 5 mM D-glucose primed HMC showed a dose-dependent migration towards MCP-1 that was dose-dependently inhibited by pertussis toxin, the broad-spectrum chemokine antagonist vMIP-II as well as the CCR2 receptor antagonist (1–8del)MCP-1 – demonstrating functional activity of MCP-1 receptor expression in primary HMC. In accordance with the downregulatory effects of 30 mM D-glucose on CCR2 mRNA expression no migratory response towards MCP-1 was observed under these conditions. The additional proinflammatory stimulus TNFα increased MCP-1 protein production in 30 as compared to 5 mM D-glucose primed HMC (2,194 ± 568 vs. 1,422 ± 379 pg MCP-1/104 cells × ml in 30 vs. 5 mM D-glucose primed HMC +24 h TNFα 500 U/ml, p = 0.002). However, this was not associated with an increased MCP-1 mRNA transcription. The 30 mM D-glucose induced downregulation of CCR2 mRNA expression was prevented in the presence of TNFα. Conclusion: High ambient glucose does not affect mesangial MCP-1 release and decreases its CCR2 receptor expression. However, in the presence of an inflammatory stimulus these effects of high glucose are reversed and an autocrine pathway of MCP-1 develops in mesangial cells.

Keywords:
Monocyte chemoattractant protein-1, CCR2, Glucose, Mesangial cell
1.
Furuta T, Saito T, Ootaka T, Soma J, Obara K, Abe K, Yoshinaga K: The role of macrophages in diabetic glomerulosclerosis. Am J Kidney Dis 1993;21:480–485.
2.
Rovin BH, Doe N, Tan LC: Monocyte chemoattractant protein-1 levels in patients with glomerular disease. Am J Kidney Dis 1996;27:640–646.
3.
Banba N, Nakamura T, Matsumura M, Kuroda H, Hattori Y, Kasai K: Possible relationship of monocyte chemoattractant protein-1 with diabetic nephropathy. Kidney Int 2000;58:684–690.
4.
Schwarz M, Radeke HH, Resch K, Uchiechowski P: Lymphocyte-derived cytokines induce sequential expression of monocyte- and T cell-specific chemokines in human mesangial cells. Kidney Int 1997;52:1521–1531.
5.
Ihm CG: Monocyte chemotactic peptide in diabetic nephropathy. Kidney Int 1997;52:S20-S22.
6.
Rovin BH, Tan LC: LDL stimulates mesangial fibronectin production and chemoattractant expression. Kidney Int 1993;43:218–225.
7.
Grandaliano G, Valente AJ, Abboud HE: A novel biologic activity of thrombin: Stimulation of monocyte chemotactic protein production. J Exp Med 1994;179:1737–1741.
8.
Sallusto F, Lanzavecchia A, Mackay CR: Chemokines and chemokine receptors in T-cell priming and Th1/Th2-mediated responses. Immunol Today 1998;19:568–574.
9.
Frade JMR, Mellado M, del Real G, Gutierrez-Ramos JC, Lind P, Martinez AC: Characterization of the CCR2 chemokine receptor: Functional CCR2 receptor expression in B cells. J Immunol 1997;159:5576–5584.
10.
Hayes IM, Jordan NJ, Towers S, Smith G, Paterson JR, Earnshaw JJ, Roach AG, Westwick J, Williams RJ: Human vascular smooth muscle cells express receptors for CC chemokines. Arterioscler Thromb Vasc Biol 1998;18:397–403.
11.
Salcedo R, Ponce ML, Young HA, Wasserman K, Ward JM, Kleinman HK, Oppenheim JJ, Murphy WJ: Human endothelial cells express CCR2 and respond to MCP-1: Direct role of MCP-1 in angiogenesis and tumor progression. Blood 2000;96:34–40.
12.
Weber KS, Nelson PJ, Grone HJ, Weber C: Expression of CCR2 by endothelial cells: Implications for MCP-1 mediated wound injury repair and in vivo inflammatory activation of endothelium. Arterioscler Thromb Vasc Biol 1999;19:2085–2093.
13.
Hogaboam CM, Bone-Larson CL, Lipinski S, Lukacs NW, Chensue SW, Strieter RM, Kunkel SM: Differential Monocyte chemoattractant protein-1 and chemokine receptor 2 expression by murine lung fibroblasts derived from Th1- and Th2-type pulmonary granuloma models. J Immunol 1999;163:2193–2201.
14.
Gharaee-Kermani M, Denholm EM, Phan SH: Costimulation of fibroblast collagen and transforming growth factor beta 1 gene expression by monocyte chemoattractant protein-1 via specific receptors. J Biol Chem 1996;271:17779–17784.
15.
Banas B, Luckow B, Möller M, Klier C, Nelson PJ, Schadde E, Brigl M, Halevy D, Holthöffer H, Reinhart B, Schlöndorff D: Chemokine and chemokine receptor expression in an novel human mesangial cell line. J Am Soc Nephrol 1999;10:2314–2322.
16.
Radeke HH, Meier B, Topley N, Floege J, Habermehl GG, Resch K: Interleukin-1α and tumor necrosis factor-α induce oxygen radical production in mesangial cells. Kidney Int 1990;37:767–775.
17.
Radeke HH, Gessner JE, Uchiechowski P, Mägert H-J, Schmidt RE, Resch K: Intrinsic human glomerular mesangial cells can express receptors for IgG complexes (hFcγRIII-A) and the associated FcεRI γ-chain. J Immunol 1994;153:1281–1292.
18.
Alpers CE, Hudkins KL, Gown AM, Johnson RJ: Enhanced expression of ‘muscle-specific’ actin in glomerulonephritis. Kidney Int 1992;41:1134–1142.
19.
Radeke HH, Resch K: The inflammatory function of renal glomerular mesangial cells and their interaction with the cellular immune system. Clin Invest 1992;70:825–842.
20.
Floege J, Topley N, Wessel K, Kaever V, Radeke HH, Hoppe J, Kishimoto T, Resch K: Monokines and platelet-derived growth factor modulate prostanoid production in growth-arrested human mesangial cells. Kidney Int 1990;37:859–869.
21.
Radeke HH, Cross AR, Hancock JT, Jones OTG, Nakamura M, Kaever V, Resch K: Functional expression of NADPH oxidase components (α- and β-subunits of cytochrome b558 and 45-kD flavoprotein) by intrinsic human glomerular mesangial cells. J Biol Chem 1991;266:21025–21029.
22.
Francki A, Uciechowski P, Floege J, von der Ohe J, Resch K, Radeke HH: Autocrine growth regulation of human glomerular mesangial cells is primarily mediated by basic fibroblast growth factor. Am J Pathol 1994;147:1372–1382.
23.
Uciechowski P, Schwarz M, Gessner E, Schmidt RE, Resch K, Radeke HH: IFN-γ induces the high affinity Fc receptor I for IgG (CD 64) on human glomerular mesangial cells. Eur J Immunol 1998;28:2928–2935.
24.
Yamagami S, Tokuda Y, Ishii K, Tanaka H, Endo N: cDNA cloning and functional expression of a human monocyte chemoattractant protein 1 receptor. Biochem Biophys Res Commun 1994;202:1156–1162.
25.
Marchand P, Resch K, Radeke HH: Selective inhibition of monocyte chemoattractant protein-1 gene expression in human embryonal kidney cells by specific triple helix-forming oligonucleotides. J Immunol 2000;164:2070–2076.
26.
Beall CJ, Breckenridge SM, Chakravarty L, Kolattukudy PE: Expression of human monocyte chemoattractant protein-1 in the yeast Pichia pastoris. Protein Expr Purif 1998;12:145–150.
27.
Reid S, Ritchie A, Boring L, Gosling J, Cooper S, Hangoc G, Charo IF, Broxmeyer HE: Enhanced myeloid progenitor cell cycling and apoptosis in mice lacking the chemokine receptor, CCR2. Blood 1999;93:1524–1533.
28.
Ihm CG, Park JK, Hong SP, Lee TW, Cho BS, Kim MJ, Cha DR, Ha H: A high glucose concentration stimulates the expression of monocyte chemotactic peptide 1 in human mesangial cells. Nephron 1998;79:33–37.
29.
Phillips AO, Steadman R, Topley N, Williams JD: Elevated D-glucose concentrations modulate TGF-β1 synthesis by human cultured renal proximal tubular epithelial cells: The permissive role of platelet-derived growth factor. Am J Pathol 1995;147:362–373.
30.
Phillips AO, Topley N, Steadman R, Morrisey K, Williams JD: Induction of TGF-β1 synthesis in D- glucose primed human proximal tubular epithelial cells by IL-1β and TNFα. Kidney Int 1996;50:1546–1554.
31.
Ha H, Lee HB: Effect of high glucose on peritoneal mesothelial cell biology. Perit Dial Int 2000;20:S15-S18.
32.
Uesugi N, Sakata N, Horiuchi S, Nagai R, Takeya M, Meng J, Saito T, Takebayash S: Glycoxidation-modified macrophages and lipid peroxidation product are associated with the progression of human diabetic nephropathy. Am J Kidney Dis 2001;38:1016–1025.
33.
Young BA, Johnson RJ, Alpers CE, Eng CE, Ghordon K, Floege J, Couser W: Cellular events in the evolution of experimental diabetic nephropathy. Kidney Int 1995;47:935–944.
34.
Sassy-Prigent C, Heudes D, Mandet C, Belair MF, Michel O, Perderau B, Bariety J, Bruneval P: Early glomerular macrophage recruitment in streptozotocin-induced diabetic rats. Diabetes 2000;49:466–475.
35.
Kato S, Luyckx VA, Ots M, Lee KW, Ziai F, Troy JL, Brener BM, MacKenzie HS: Renin-angiotensin blockade lowers MCP-1 expression in diabetic rats. Kidney Int 1999;56:1037–1048.
36.
Weber C, Draude G, Weber KS, Wubert J, Lorenz RL, Weber PC: Downregulation by tumor necrosis factor-alpha of monocyte CCR2 expression and monocyte chemotactic-1-induced transendothelial migration is antagonized by oxidized low-density lipoprotein: A potential mechanism of monocyte retention in atherosclerotic lesions. Atherosclerosis 1999;145:115–123.
37.
Xu L, Khandaker MH, Barlic J, Ran L, Borja ML, Madrenas J, Rahimpour R, Chen K, Mitchell G, Tan CM, DeVries M, Feldman RD, Kelvin DJ: Identification of a novel mechanism for endotoxin-mediated down-modulation of CC chemokine receptor expression. Eur J Immunol 2000;30:227–235.
38.
Sica A, Caccani A, Botazzi B, Bernasconi S, Allavena P, Gaetano B, Fei F, LaRosa G, Scotton C, Balkwill F, Mantovani A: Defective expression of the monocyte chemotactic protein-1 receptor CCR2 in macrophages associated with human ovarian carcinoma. J Immunol 2000;164:733–738.
39.
Premack BA, Schall TJ: Chemokine receptors: Gateways to inflammation and infection. Nat Med 1996;2:1174–1178.
40.
Kledal TN, Rosenkilde MM, Coulin F, Simmons G, Johnsen AH, Alouani S, Power CA, Luttichau HR, Gerstoft J, Clapham PR, Clark-Lewis I, Wells TNC, Schwartz TW: A broad-spectrum chemokine antagonist encoded by Kaposi’s sarcoma-associated herpesvirus. Science 1997;277:1656–1659.
41.
Jarnagin K, Grunberger D, Mulkins M, Wong B, Hemmerich S, Paavola C, Bloom A, Bhakta S, Diehl F, Freedman R, McCarley D, Polsky I, Ping-Tsou A, Kosaka A, Handel TM: Identification of surface residues of the monocyte chemotactic protein 1 that affect signaling through the receptor CCR2. Biochemistry 1999;38:16167–16177.
42.
Aragay AM, Mellado M, Frade JMR, Martin AM, Jimenez MC, Martinez-AC, Mayor F Jr: Monocyte chemoattractant protein-1-induced CCR2B receptor desensitization mediated by the G protein-coupled receptor kinase 2. Proc Natl Acad Sci USA 1998;95:2985–2990.
43.
Bird JE, Giancarli MR, Kurihara T, Kowala MC, Valentine MT, Gitlitz PH, Pandya DG, French MH, Durham SK: Increased severity of glomerulonephritis in C-C chemokine receptor 2 knockout mice. Kidney Int 2000;57:129–136.
© 2002 S. Karger AG, Basel
2002
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.
You do not currently have access to this content.