Abstract
Background: Hyperglycemia directly contributes to the development of diabetic nephropathy. Nitric oxide (NO), a potent endothelium-derived vasodilator, has been suggested to participate in the regulation of renal blood flow, glomerular filtration rate, and mesangial matrix accumulation. Human vascular endothelial cells are known to exhibit functional heterogeneity, this prompted us to do the first study of NO bioavailability in human glomerular endothelial cells (HGECs), in response to high glucose exposure. Methods: NO release was examined by detecting nitrite generation by the Griess assay in HGECs exposed to control-level (5.5 mM) and high-level (15, 30 and 60 mM) glucose solutions at various time periods (24, 48 and 72 h) in the presence or absence of L-arginine (1 mM), or superoxide dismutase (SOD) (250 U/ml). In addition, we evaluated the effect of glucose on the expression of endothelial nitric oxide synthase (eNOS) in HGECs by Western blotting. Results: Final levels of nitrite generated in HGECs were reduced significantly, in a time- and concentration-dependent manner, after high glucose exposure. However, Western blot analysis revealed that eNOS protein expression was significantly upregulated at 12 h after exposure to high glucose concentrations (30 mM), reaching a peak at 48 h (twofold increase over baseline levels). The inhibitory effect of high glucose on NO production was restored by the addition of SOD. Addition of L-arginine (1 mM) to external media also reversed the inhibitory effect of high glucose on NO production of HGECs as well. Conclusions: The present study demonstrated that high glucose increased eNOS protein expression, but decreased NO release finally. Decreased NO bioavailability seems to be associated with overproduction of superoxide and L-arginine deficiency. These findings provide an important clue in clarifying the molecular basis of the mechanisms by which elevated glucose leads to an imbalance between NO and superoxide, resulting in impaired endothelial function. In addition, restoration of NO function by both administration of L-arginine and adequate intake of antioxidants suggests a potential supportive treatment for patients with diabetic nephropathy.
Journal Section:
Original Paper
References
1.
Jacobs C, Selwood NH: Renal replacement therapy for end-stage renal failure in France: Current status and evolutive trends over the last decade. Am J Kidney Dis 1995;25:188–195.
[PubMed]
2.
Ratner RE: Type 2 diabetes mellitus: The grand overview. Diabet Med 1998;15:S4–S7.
3.
Tesfamariam B, Brown ML, Deykin D, Cohen RA: Elevated glucose promotes generation of endothelium-derived vasoconstrictor prostanoids in rabbit aorta. J Clin Invest 1990;85:929–932.
[PubMed]
4.
Mogensen CE: Microalbuminuria, blood pressure and diabetic renal disease: Origin and development of ideas. Diabetologia 1999;42:263–285.
[PubMed]
5.
Ruggenenti P, Remuzzi G: Nephropathy of type-2 diabetes mellitus. J Am Soc Nephrol 1998;9:2157–2169.
[PubMed]
6.
Laakso M: Hyperglycemia and cardiovascular disease in type 2 diabetes. Diabetes 1999;48:937–942.
[PubMed]
7.
Sugimoto H, Shikata K, Matsuda M, Kushiro M, Hayashi Y, Hiragushi K, Wada J, Makino H: Increased expression of endothelial cell nitric oxide synthase (ecNOS) in afferent and glomerular endothelial cells is involved in glomerular hyperfiltration of diabetic nephropathy. Diabetologia 1998;41:1426–1434.
[PubMed]
8.
Lorenzi M, Cagliero E: Pathobiology of endothelial and other vascular cells in diabetes mellitus. Call for data. Diabetes 1991;40:653–659.
[PubMed]
9.
Honing ML, Morrison PJ, Banga JD, Stroes ES, Rabelink TJ: Nitric oxide availability in diabetes mellitus. Diabetes Metab Rev 1998;14:241–249.
[PubMed]
10.
Loscalzo J: Nitric oxide and vascular disease. N Engl J Med 1995;333:251–253.
11.
Schwartz D, Mendonca M, Schwartz I, Xia Y, Satriano J, Wilson CB, Blantz RC: Inhibition of constitutive nitric oxide synthase (NOS) by nitric oxide generated by inducible NOS after lipopolysaccharide administration provokes renal dysfunction in rats. J Clin Invest 1997;100:439–448.
[PubMed]
12.
Murakami S, Morioka T, Nakagawa Y, Suzuki Y, Arakawa M, Oite T: Expression of adhesion molecules by cultured human glomerular endothelial cells in response to cytokines: Comparison to human umbilical vein and dermal microvascular endothelial cells. Microvasc Res 2001;62:383–391.
13.
Geiger M, Stone A, Mason SN, Oldham KT, Guice KS: Differential nitric oxide production by microvascular and macrovascular endothelial cells. Am J Physiol 1997;273:L275–L281.
14.
Yao J, Morioka T, Li B, Oite T: Endothelin is a potent inhibitor of matrix metalloproteinase-2 secretion and activation in rat mesangial cells. Am J Physiol Renal Physiol 2001;280:F628–F635.
15.
Li B, Yao J, Morioka T, Oite T: Nitric oxide increases albumin permeability of isolated rat glomeruli via a phosphorylation-dependent mechanism. J Am Soc Nephrol 2001;12:2616–2624.
[PubMed]
16.
Graier WF, Posch K, Fleischhacker E, Wascher TC, Kostner GM: Increased superoxide anion formation in endothelial cells during hyperglycemia: An adaptive response or initial step of vascular dysfunction? Diabetes Res Clin Pract 1999;45:153–160.
17.
Rosen P, Nawroth PP, King G, Moller W, Tritschler HJ, Packer L: The role of oxidative stress in the onset and progression of diabetes and its complications: A summary of a Congress Series sponsored by UNESCO-MCBN, the American Diabetes Association and the German Diabetes Society. Diabetes Metab Res Rev 2001;17:189–212.
18.
Nishikawa T, Edelstein D, Du XL, Yamagishi S, Matsumura T, Kaneda Y, Yorek MA, Beebe D, Oates PJ, Hammes HP, Giardino I, Brownlee M: Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature 2000;404:787–790.
[PubMed]
19.
Evans JL, Goldfine ID, Maddux BA, Grodsky GM: Are oxidative stress-activated signaling pathways mediators of insulin resistance and β-cell dysfunction? Diabetes 2003;52:1–8.
20.
Abid MR, Tsai JC, Spokes KC, Deshpande SS, Irani K, Aird WC: Vascular endothelial growth factor induces manganese-superoxide dismutase expression in endothelial cells by a Rac1-regulated NADPH oxidase-dependent mechanism. FASEB J 2001;15:2548–2550.
[PubMed]
21.
Diabetes Control and Complications Trial Research Group: The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. N Engl J Med 1993;329:977–986.
[PubMed]
22.
Oyanagi-Tanaka Y, Yao J, Wada Y, Morioka T, Suzuki Y, Gejyo F, Arakawa M, Oite T: Real-time observation of hemodynamic changes in glomerular aneurysms induced by anti-Thy-1 antibody. Kidney Int 2001;59:252–259.
23.
Wada Y, Morioka T, Oyanagi-Tanaka Y, Yao J, Suzuki Y, Gejyo F, Arakawa M, Oite T: Impairment of vascular regeneration precedes progressive glomerulosclerosis in anti-Thy-1 glomerulonephritis. Kidney Int 2002;61:432–443.
[PubMed]
24.
Goligorsky MS, Chen J, Brodsky S: Endothelial cell dysfunction leading to diabetic nephropathy: Focus on nitric oxide. Hypertension 2001;37:744–748.
[PubMed]
25.
Giugliano D, Marfella R, Coppola L, Verrazzo G, Acampora R, Giunta R, Nappo F, Lucarelli C, D’Onofrio F: Vascular effects of acute hyperglycemia in humans are reversed by L-arginine. Evidence for reduced availability of nitric oxide during hyperglycemia. Circulation 1997;95:1783–1790.
[PubMed]
26.
Du X, Stocklauser-Farber K, Rosen P: Generation of reactive oxygen intermediates, activation of NF-κB, and induction of apoptosis in human endothelial cells by glucose: Role of nitric oxide synthase? Free Radic Biol Med 1999;27:752–763.
[PubMed]
27.
Derubertis FR, Craven PA: Activation of protein kinase C in glomerular cells in diabetes. Mechanisms and potential links to the pathogenesis of diabetic glomerulopathy. Diabetes 1994;43:1–8.
[PubMed]
28.
Brodsky SV, Morrishow AM, Dharia N, Gross SS, Goligorsky MS: Glucose scavenging of nitric oxide. Am J Physiol Renal Physiol 2001;280:F480–F486.
29.
Craven PA, Studer RK, DeRubertis FR: Impaired nitric oxide release by glomeruli from diabetic rats. Metabolism 1995;44:695–698.
[PubMed]
30.
Govers R, Rabelink TJ: Cellular regulation of endothelial nitric oxide synthase. Am J Physiol Renal Physiol 2001;280:F193–F206.
31.
Pieper GM, Peltier BA: Amelioration by L-arginine of a dysfunctional arginine/nitric oxide pathway in diabetic endothelium. J Cardiovasc Pharmacol 1995;25:397–403.
[PubMed]
32.
Pieper GM, Siebeneich W, Moore-Hilton G, Roza AM: Reversal by L-arginine of a dysfunctional arginine/nitric oxide pathway in the endothelium of the genetic diabetic BB rat. Diabetologia 1997;40:910–915.
[PubMed]
33.
Huang S, Fu P, Li X, Zhang X: L-Arginine and nitric oxide have effects on glomerulus hyperperfusion of early diabetic rats (in Chinese). Hua Xi Yi Ke Da Xue Xue Bao 1997;28:251–254.
[PubMed]
34.
Knowles RG, Moncada S: Nitric oxide synthases in mammals. Biochem J 1994;298:249–258.
[PubMed]
35.
Palmer RM, Ashton DS, Moncada S: Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 1988;333:664–666.
[PubMed]
36.
Wever RM, van Dam T, van Rijn HJ, de Groot F, Rabelink TJ: Tetrahydrobiopterin regulates superoxide and nitric oxide generation by recombinant endothelial nitric oxide synthase. Biochem Biophys Res Commun 1997;237:340–344.
[PubMed]
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