Background/Aim: Increased nitric oxide (NO) generation and action have been suggested to be associated with glomerular hyperfiltration and increased vascular permeability early in diabetes. However, previous studies have primarily focused on the constitutive nitric oxide synthase (cNOS) pathway present in endothelial cells, and the role of the inducible NOS (iNOS) pathway in diabetic nephropathy has remained unclear. This study examined whether high glucose modulates NO synthesis by the iNOS pathway in rat mesangial cells. In addition, the effect of inhibition of the iNOS pathway on fibronectin production was determined to examine the role of the iNOS pathway in high glucose-induced extracellular expansion by mesangial cells. Methods: NO synthesis by the iNOS pathway was evaluated by nitrite and iNOS mRNA and protein productions. The effects of protein kinase C (PKC) inhibitor and aldose reductase inhibitor on the iNOS mRNA expression and aminoguanidine, a relatively specific inhibitor of the iNOS on fibronectin protein production were examined. Results: High 30 mM glucose concentration led to significant increases in nitrite production of rat mesangial cells upon stimulation with lipopolysaccharide (LPS) plus interferon-γ (IFN-γ) compared with control 5.6 mM glucose concentration. Mesangial iNOS mRNA expression and protein production also increased significantly in response to high glucose. The addition of calphostin C, a PKC inhibitor, and 6-bromo-1,3-dioxo-1H-benz[d,e]isoquinoline-2(3H)-acetic acid, an aldose reductase inhibitor, significantly suppressed the enhancement of iNOS mRNA expression in high glucose concentration. High glucose also significantly increased fibronectin protein production of mesangial cells upon stimulation with LPS plus IFN-γ compared to control glucose. Aminoguanidine reversed this high glucose-induced fibronectin production at dose inhibiting iNOS mRNA expression. Conclusions: These results indicate that high glucose enhances cytokine-induced NO production by rat mesangial cells, and that the activation of PKC and aldose reductase pathway may play a role in this enhancement. In addition, high glucose-induced NO production by the iNOS pathway may promote extracellular matrix accumulation by mesangial cells under certain condition.

1.
Tilton RG, Chang K, Pugliese G, et al: Prevention of hemodynamic and vascular albumin filtration changes in diabetic rats by aldose reductase inhibitors. Diabetes 1989;38:1258–1270.
2.
Vlassara H, Bucala R: Recent progress in advanced glycation and diabetic vascular disease: Role of advanced glycation end product receptors. Diabetes 1996;45(suppl 3):S65–S66.
3.
Diederich D: NO in diabetic nephropathy in nitric oxide and the kidney; in Goligorsky MS, Gross SS (eds): Physiology and Pathophysiology. New York, Chapman & Hall, 1997, pp 349–367.
4.
Lee HB, Ha H: Introduction to experimental approaches to diabetic nephropathy. Kidney Int 1997;52(suppl 60):S1–S2.
5.
Kolm-Litty V, Sauer U, Nerlich A, Lehmann R, Schleicher ED: High glucose-induced transforming growth factor-beta1 production is mediated by the hexosamine pathway in porcine glomerular mesangial cells. J Clin Invest 1998;101:160–169.
6.
Baynes JW, Thorpe SR: Role of oxidative stress in diabetic complications: A new perspective on an old paradigm. Diabetes 1999;48:1–9.
7.
Bank N, Aynedjian HS: Role of EDRF (nitric oxide) in diabetic renal hyperfiltration. Kidney Int 1993;43:1306–1312.
8.
Tolins JP, Shultz PJ, Raij L, Brown DM, Mauer SM: Abnormal renal hemodynamic response to reduced renal perfusion pressure in diabetic rats: Role of NO. Am J Physiol 1993;265:F886–895.
9.
Craven PA, Studer RK, DeRubertis FR: Impaired nitric oxide-dependent cyclic guanosine monophosphate generation in glomeruli from diabetic rats: Evidence for protein kinase C-mediated suppression of the cholinergic response. J Clin Invest 1994;93:311–320.
10.
Craven PA, Studer RK, DeRubertis FR: Impaired nitric oxide release by glomeruli from diabetic rats. Metabolism 1995;44:695–698.
11.
Craven PA, DeRubertis FR, Melhem M: Nitric oxide in diabetic nephropathy. Kiney Int 1997;52(suppl 60):S46–S53.
12.
Sharma K, Danoff TM, DePiero A, Ziyadeh FN: Enhanced expression of inducible nitric oxide synthase in murine macrophages and glomerular mesangial cells by elevated glucose levels: Possible mediation via protein kinase C. Biochem Biophys Res Commun 1995;207:80–88.
13.
Trachtman H, Futterweit S, Crimmins DL: High glucose inhibits nitric oxide production in cultured rat mesangial cells. J Am Soc Nephrol 1997;8:1276–1282.
14.
Narita I, Border WA, Ketteler M, Noble NA: Nitric oxide mediates immunologic injury to kidney mesangium in experimental glomerulonephritis. Lab Invest 1995;72:17–24.
15.
Yang CW, Yu CC, Ko YC, Huang CC: Aminoguanidine reduces glomerular inducible nitric oxide synthase (iNOS) and transforming growth factor-beta1 (TGF-β1) mRNA expression and diminishes glomerulosclerosis in NZB/W F1 mice. Clin Exp Immunol 1998;113:258–264.
16.
Oh JH, Ha H, Yu MR, Lee HB: Sequential effects of high glucose on mesangial cell transforming growth factor-β1 and fibronectin synthesis. Kidney Int 1998;54:1872–1878.
17.
Ding AH, Nathan CF, Stuehr DJ: Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages: Comparison of activating cytokines and evidence for independent production. J Immunol 1988;141:2407–2412.
18.
Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–159.
19.
Graier WF, Simecek S, Kukovetz WR, Kostner GM: High D-glucose-induced changes in endothelial Ca2+/EDRF signaling are due to generation of superoxide anions. Diabetes 1996;45:1386–1395.
20.
Calver AC, Collier JG, Vallance PJT: Inhibition and stimulation of nitric oxide synthesis in the human forearm arterial bed of patients with insulin dependent diabetes. J Clin Invest 1992;90:2548–2554.
21.
McVeigh GE, Brennan GM, Johnston GD, et al: Impaired endothelium dependent and independent vasodilation in patients with type 2 diabetes mellitus. Diabetologia 1992;35:771–776.
22.
Johnstone MT, Creager SJ, Scales KM, Cusco JA, Lee BK, Creager MA: Impaired endothelium dependent vasodilation in patients with insulin dependent diabetes mellitus. Circulation 1993;88:2510–2516.
23.
Williams SB, Cusco JA, Roddy MA, Johnstone MT, Creager MA: Impaired nitric oxide mediated vasodilation in non insulin dependent diabetes. J Am Coll Cardiol 1996;27:567–574.
24.
Wang YX, Brooks DP, Edwards RM: Attenuated glomerular cGMP production and renal vasodilation in streptozotocin-induced diabetic rats. Am J Physiol 1993;264:R952–956.
25.
Nakamura T, Fukui M, Ebihara I, et al: mRNA expression of growth factors in glomeruli from diabetic rats. Diabetes 1993;42:450–456.
26.
Young B, Johnson R, Alpers C, Eng E, Floege J, Couser W: Mesangial cell proliferation precedes development of glomerulosclerosis in experimental diabetic nephropathy (abstract). J Am Soc Nephrol 1992;3:770.
27.
Vodovotz Y, Bogdan C, Paik J, Xie QW, Nathan C: Mechanisms of suppression of macrophage nitric oxide release by transforming growth factor beta. J Exp Med 1993;178:605–613.
28.
Trachtman H, Koss I, Bogart M, et al: High glucose enhances growth factor-stimulated nitric oxide production by cultured rat mesangial cells. Res Commun Mol Pathol Pharmacol 1998;100:213–225.
29.
Xie QW, Cho HJ, Calaycay J, et al: Cloning and characterization of inducible nitric oxide synthase from mouse macrophages. Science 1992;256:225–228.
30.
Marsden PA, Ballermann BJ: Tumor necrosis factor alpha activates soluble cyclase in bovine glomerular mesangial cells via an L-arginine dependent mechanism. J Exp Med 1990;172:1843–1852.
31.
Hortelano S, Genaro AM, Bosca L: Phorbol esters induce nitric oxide synthase and increase arginine influx in cultured peritoneal macrophages. FEBS Lett 1993;320:135–139.
32.
Bosca L, Lazo PA: Induction of nitric oxide release by MRC OX-44 (anti-CD53) through a protein kinase C-dependent pathway in rat macrophages. J Exp Med 1994;179:1119–1126.
33.
Okuda Y, Kawashima K, Suzuki S, et al: Restoration of nitric oxide production by aldose reductase inhibitor in human endothelial cells cultured in high-glucose medium. Life Sci 1997;60:PL53–56.
34.
Sochor M, Baquer NZ, McLean P: Regulation of pathways of glucose metabolism in kidney: The effect of experimental diabetes on the activity of the pentose phosphate pathway and the glucuronate-xylulose pathway. Arch Biochem Biophys 1979;198:632–646.
35.
Williamson JR, Chang K, Frangos M, et al: Hyperglycemic pseudohypoxia and diabetic complications. Diabetes 1993;42:801–813.
36.
Kapor-Drezgic J, Zhou X, Babazono T, Dlugosz JA, Hohman T, Whiteside C: Effect of high glucose on mesangial cell protein kinase C-δ and -ε is polyol pathway dependent. J Am Soc Nephrol 1999;10:1193–1203.
37.
Furusu A, Miyazaki M, Abe K, et al: Expression of endothelial and inducible nitric oxide synthase in human glomerulonephritis. Kidney Int 1998;53:1760–1768.
38.
Chatziantoniou C, Boffa JJ, Ardaillou R, Dussaule JC: Nitric oxide inhibition induces early activation of type I collagen gene in renal resistance vessels and glomeruli in transgenic mice: Role of endothelin. J Clin Invest 1998;101:2780–2789.
39.
Trachtman H, Futterweit S, Singhal P: Nitric oxide modulates the synthesis of extracellular matrix proteins in cultured rat mesangial cells. Biochem Biophys Res Commun 1995;207:120–125.
40.
Craven PA, Studer RK, Felder J, Phillips S, DeRubertis FR: Nitric oxide inhibition of transforming growth factor-β and collagen synthesis in mesangial cells. Diabetes 1997;46:671–681.
41.
Trachtman H, Futterweit S, Garg P, Reddy K, Singhal PC: Nitric oxide stimulates the activity of a 72-kDa neutral matrix metalloproteinase in cultured rat mesangial cells. Biochem Biophys Res Commun 1996;218:704–708.
42.
Giardino I, Fard AK, Hatchell DL, Brownlee M: Aminoguanidine inhibits reactive oxygen species formation, lipid peroxidation, and oxidant-induced apoptosis. Diabetes 1998;47:1114–1120.
43.
Studer RK, Craven PA, DeRubertis FR: Antioxidant inhibition of protein kinase C-signaled increase in transforming growth factor-beta in mesangial cells. Metabolism 1997;46:918–925.
44.
Trachtman H, Futterweit S, Bienkowski RS: Taurine prevents glucose-induced lipid peroxidation and increased collagen production in cultured rat mesangial cells. Biochem Biophys Res Commun 1993;191:759–765.
45.
Trachtman H: Vitamin E prevents glucose-induced lipid peroxidation and increased collagen production in cultured rat mesangial cells. Microvasc Res 1994;47:232–239.
46.
Ha H, Lee SH, Kim KH: Effects of rebamipide in a model of experimental diabetes on the synthesis of transforming growth factor-β and fibronectin, and lipid peroxidation induced by high glucose in cultured mesangial cells. J Pharmacol Exp Ther 1997;281:1457–1462.
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.