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Diabetologia
Published in: Diabetologia 6/2003

01-06-2003 | Article

Intercellular signalling within vascular cells under high D-glucose involves free radical-triggered tyrosine kinase activation

Authors: G. Schaeffer, S. Levak-Frank, M. M. Spitaler, E. Fleischhacker, V. E. Esenabhalu, A. H. Wagner, M. Hecker, W. F. Graier

Published in: Diabetologia | Issue 6/2003

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Abstract

Aims/hypothesis

Diabetes mellitus is associated with endothelial dysfunction in human arteries due to the release of superoxide anions (·O2 –) that was found to occur predominantly in smooth muscle cells (SMC). This study was designed to elucidate the impact of high glucose concentration mediated radical production in SMC on EC. Pre-treatment of vascular SMC with increased D-glucose enhanced release of ·O2 –.

Methods

Microscope-based analyses of intracellular free Ca2+ concentration (fura-2), immunohistochemistry (f-actin) and tyrosine kinase activity were performed. Furthermore, RT-PCR and Western blots were carried out.

Results

Interaction of EC with SMC pre-exposed to high glucose concentration yielded changes in endothelial Ca2+ signalling and polymerization of f-actin in a concentration-dependent and superoxide dismutase (SOD) sensitive manner. This interaction activated endothelial tyrosine kinase(s) but not NFκB and AP-1, while SOD prevented tyrosine kinase stimulation but facilitated NFκB and AP-1 activation. Erbstatin, herbimycin A and the src family specific kinase inhibitor PP-1 but not the protein kinase C inhibitor GF109203X prevented changes in endothelial Ca2+ signalling and cytoskeleton organization induced by pre-exposure of SMC to high glucose concentration. Adenovirus-mediated expression of kinase-inactive c-src blunted the effect of pre-exposure of SMC to high glucose concentration on EC.

Conclusions/interpretation

These data suggest that SMC-derived ·O2 – alter endothelial cytoskeleton organization and Ca2+ signalling via activation of c-src. The activation of c-src by SMC-derived radicals is a new concept of the mechanisms underlying vascular dysfunction in diabetes.
Literature
1.
Cohen R (1993) Dysfunction of vascular endothelium in diabetes mellitus. Circulation 87 [Suppl V]:V67–V76
2.
Kirpichnikov D, Sowers JR (2001) Diabetes mellitus and diabetes-associated vascular disease. Trends Endocrinol Metab 12:225–230CrossRefPubMed
3.
Schaeffer G, Wascher TC, Kostner GM, Graier WF (1999) Alterations in platelet Ca2+ signalling in diabetic patients is due to increased formation of superoxide anions and reduced nitric oxide production. Diabetologia 42:167–176CrossRefPubMed
4.
Srivastava AK, St-Louis J (1997) Smooth muscle contractility and protein tyrosine phosphorylation. Mol Cell Biochem 176:47–51CrossRefPubMed
5.
Srivastava AK (2002) High glucose-induced activation of protein kinase signaling pathways in vascular smooth muscle cells: a potential role in the pathogenesis of vascular dysfunction in diabetes. Int J Mol Med 9:85–89PubMed
6.
Fleischhacker E, Esenabalu VE, Spitaler M, Holzmann S, Skrabal F, Koidl B, Kostner GM, Graier WF (1999) Human diabetes is associated with hyperreactivity of vascular smooth muscle cells due to altered subcellular Ca2+ distribution. Diabetes 48:1323–1330PubMed
7.
Spitaler MM, Graier WF (2002) Vascular targets of redox signalling in diabetes mellitus. Diabetologia 45:476–494CrossRef
8.
Pieper GM (1998) Review of alterations in endothelial nitric oxide production in diabetes: protective role of arginine on endothelial dysfunction. Hypertension 31:1047–1060PubMed
9.
Idris IG, Donnelly R (2001) Protein kinase C activation: isozyme-specific effects on metabolism and cardiovascular complications in diabetes. Diabetologia 44:659–673CrossRefPubMed
10.
Graier WF, Simecek S, Kukovetz WR, Kostner GM (1996) High-D-glucose-induced changes in endothelial Ca2+/EDRF signaling are due to generation of superoxide anions. Diabetes 45:1386–1395PubMed
11.
Graier WF, Wascher TC, Lackner L, Toplak H, Krejs GJ, Kukovetz WR (1993) Exposure to elevated D-glucose concentrations modulates vascular endothelial cell vasodilatatory response. Diabetes 42:1497–1505PubMed
12.
Graier WF, Hoebel BG, Paltauf-Doburzynska J, Kostner GM (1998) Effects of superoxide anions on endothelial Ca2+ signaling pathways. Arterioscler Thromb Vasc Biol 18:1470–1479PubMed
13.
Steinbrecher UP (1988) Role of superoxide in endothelial-cell modification of low-density lipoproteins. Biochim Biophys Acta 959:20–30CrossRefPubMed
14.
Frieden M, Malli R, Samardzija M, Demaurex N, Graier WF (2002) Subplasmalemmal endoplasmic reticulum controls KCa channel activity upon stimulation with a moderate histamine concentration in a human umbilical vein endothelial cell line. J Physiol 540:73–84CrossRefPubMed
15.
Krzesz R, Wagner AH, Cattaruzza M, Hecker M (1999) Cytokine-inducible CD40 gene expression in vascular smooth muscle cells is mediated by nuclear factor kappaB and signal transducer and activation of transcription-1. FEBS Lett 453:191–196PubMed
16.
Hoebel BG, Graier WF (1998) 11,12-Epoxyeicosatrienoic acid stimulates tyrosine kinase activity in porcine aortic endothelial cells. Eur J Pharmacol 346:115–117CrossRefPubMed
17.
Kurokawa K, Mochizuki N, Ohba Y, Mizuno H, Miyawaki A, Matsuda M (2001) A pair of fluorescent resonance energy transfer-based probes for tyrosine phosphorylation of the CrkII adaptor protein in vivo. J Biol Chem 276:31305–31310CrossRefPubMed
18.
Okuda M, Takahashi M, Suero J et al. (1999) Shear stress stimulation of p130(cas) tyrosine phosphorylation requires calcium-dependent c-Src activation. J Biol Chem 274:26803–26809CrossRefPubMed
19.
Graier WF, Myers PR, Adams HR, Parker JL (1994) E. coli endotoxin inhibits agonist-mediated cytosolic calcium mobilization and nitric oxide biosynthesis in cultured endothelial cells. Circ Res 75:659–668PubMed
20.
Doan TN, Gentry DL, Taylor AA, Elliott SJ (1994) Hydrogen peroxide activates agonist-sensitive Ca2+-flux pathways in canine venous endothelial cells. Biochem J 297:209–215PubMed
21.
Elliott SJ, Doan TN (1993) Oxidant stress inhibits the store-dependent Ca2+-influx pathway of vascular endothelial cells. Biochem J 292:385–393PubMed
22.
Koliwad SK, Kunze DL, Elliott SJ (1996) Oxidant stress activates a non-selective cation channel responsible for membrane depolarization in calf vascular endothelial cells. J Physiol 491:1–12PubMed
23.
Zhao B, Ehringer WD, Dierichs R, Miller FN (1997) Oxidized low-density lipoprotein increases endothelial intracellular calcium and alters cytoskeletal f-actin distribution. Eur J Clin Invest 27:48–54PubMed
24.
Kamata K, Sugiura M, Kasuya Y (1995) Decreased Ca2+ influx into the endothelium contributes to the decrease in endothelium-dependent relaxation in the aorta of streptozotocin-induced diabetic mice. Res Commun Mol Pathol Pharmacol 90:69–74PubMed
25.
Kamata K, Nakajima M (1998) Ca2+ mobilization in the aortic endothelium in streptozotocin-induced diabetic and cholesterol-fed mice. Br J Pharmacol 123:1509–1516PubMed
26.
Pieper GM, Dondlinger L (1997) Glucose elevations alter bradykinin-stimulated intracellular calcium accumulation in cultured endothelial cells. Cardiovasc Res 34:169–178CrossRefPubMed
27.
Graier WF, Posch K, Wascher TC, Kostner GM (1997) Role of superoxide anions in changes of endothelial vasoactive response during acute hyperglycemia. Horm Met Res 29:622–626
28.
Graier WF, Posch K, Fleischhacker E, Wascher TC, Kostner GH (1999) Increased superoxide anion formation in endothelial cells during hyperglycemia: an adaptive response or initial step of vascular dysfunction? Diabet Res Clin Pract 45:153–160
29.
Pagano PJ, Ito Y, Tornheim K, Gallop P, Tauber AI, Cohen RA (1995) An NADPH oxidase superoxide-generating system in the rabbit aorta. Am J Physiol 268:H2274–H2280PubMed
30.
Griendling KK, Alexander RW (1996) Endothelial control of the cardiovascular system: recent advances. Faseb J 10:283–292PubMed
31.
Griendling KK, Sorescu D, Ushio-Fukai M (2000) NAD(P)H oxidase: role in cardiovascular biology and disease. Circ Res 86:494–501PubMed
32.
Mohazzab KM, Wolin MS (1994) Sites of superoxide anion production detected by lucigenin in calf pulmonary artery smooth muscle. Am J Physiol 267:L815–L822PubMed
33.
Mohazzab KM, Kaminski PM, Wolin MS (1994) NADH oxidoreductase is a major source of superoxide anion in bovine coronary artery endothelium. Am J Physiol 266:H2568–H2572PubMed
34.
Nishikawa T, Edelstein D, Du XL et al. (2000) Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature 404:787–790PubMed
35.
Nishikawa T, Edelstein D, Brownlee M (2000) The missing link: a single unifying mechanism for diabetic complication. Kidney Int 58 [Suppl]77:26–30
36.
Holda JR, Blatter A (1997) Capacitative calcium entry is inhibited in vascular endothelial cells by disruption of cytoskeletal microfilaments. FEBS Lett 403:191–196PubMed
37.
Rosado JA, Sage SO (2000) The actin cytoskeleton in store-mediated calcium entry. J Physiol 526:221–229PubMed
38.
Rosado JA, Jenner S, Sage SO (2000) A role for the actin cytoskeleton in the initiation and maintenance of store-mediated calcium entry in human platelets. Evidence for conformational coupling. J Biol Chem 275:7527–7533CrossRefPubMed
39.
Wang YJ, Gregory RB, Barritt GJ (2002) Maintenance of the filamentous actin cytoskeleton is necessary for the activation of store-operated Ca2+ channels, but not other types of plasma-membrane Ca2+ channels, in rat hepatocytes. Biochem J 363:117–126CrossRefPubMed
40.
Sundaresan MY, Ferrans ZX, Sulciner VJ et al. (1996) Regulation of reactive-oxygen-species generation in fibroblasts by Rac1. Biochem J 318:379–382PubMed
41.
Irani K, Xia Y, Zweier JL et al. (1997) Mitogenic signaling mediated by oxidants in Ras-transformed fibroblasts. Science 275:1649–1652CrossRefPubMed
42.
Irani K, Goldschmidt-Clermont PJ (1998) Ras, superoxide and signal transduction. Biochem Pharmacol 55:1339–1346
43.
Irani K (2000) Oxidant signaling in vascular cell growth, death, and survival : a review of the roles of reactive oxygen species in smooth muscle and endothelial cell mitogenic and apoptotic signaling. Circ Res 87:179–183PubMed
44.
Huttunen HJ, Fages C, Rauvala H (1999) Receptor for advanced glycation end products (RAGE)-mediated neurite outgrowth and activation of NF-kappaB require the cytoplasmic domain of the receptor but different downstream signaling pathways. J Biol Chem 274:19919–19924CrossRefPubMed
45.
46.
Gilbert DA, Bergel F (1964) The chemistry of xanthine oxidase. 9. An improved method of preparing the bovine milk enzyme. Biochem J 90:350–353PubMed
47.
Garcia R, Parikh NU, Saya H, Gallick GE (1991) Effect of herbimycin A on growth and pp60c-src activity in human colon tumor cell lines. Oncogene 6:1983–1989PubMed
48.
Hanke JH, Gardner JP et al. (1996) Discovery of a novel, potent, and Src family-selective tyrosine kinase inhibitor. Study of Lck- and FynT-dependent T cell activation. J Biol Chem 271:695–701PubMed
49.
Toullec D, Pianetti P, Coste H et al. (1991) The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem 266:15771–15781PubMed
50.
Inoguchi TL, Umeda P, Yu F et al.(2000) High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells. Diabetes 49:1939–1945PubMed
51.
Blanchetot C, Tertoolen LG, Hertog J den (2002) Regulation of receptor protein-tyrosine phosphatase alpha by oxidative stress. EMBO J 21:493–503CrossRefPubMed
52.
Liao DF, Jin ZG, Baas AS et al. (2000) Purification and identification of secreted oxidative stress-induced factors from vascular smooth muscle cells. J Biol Chem 275:189–196CrossRefPubMed
53.
Ceriello A, Russo P dello, Amstad P, Cerutti P (1996) High glucose induces antioxidant enzymes in human endothelial cells in culture. Evidence linking hyperglycemia and oxidative stress. Diabetes 45:471–477PubMed
Metadata
Title
Intercellular signalling within vascular cells under high D-glucose involves free radical-triggered tyrosine kinase activation
Authors
G. Schaeffer
S. Levak-Frank
M. M. Spitaler
E. Fleischhacker
V. E. Esenabhalu
A. H. Wagner
M. Hecker
W. F. Graier
Publication date
01-06-2003
Publisher
Springer-Verlag
Published in
Diabetologia / Issue 6/2003
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI
https://doi.org/10.1007/s00125-003-1091-y

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