Effect of High Glucose on the Production of Reactive Oxygen Species in Trabecular Meshwork Cells

Chang Beum Bae; Jae Woo Kim

doi:10.3341/jkos.2010.51.3.418

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Bae and Kim: Effect of High Glucose on the Production of Reactive Oxygen Species in Trabecular Meshwork Cells

Original Article

J Korean Ophthalmol Soc 2010;51(3):418-422.

Published online: 25 September 2010

DOI: https://doi.org/10.3341/jkos.2010.51.3.418

Effect of High Glucose on the Production of Reactive Oxygen Species in Trabecular Meshwork Cells

Chang Beum Bae, MD, Jae Woo Kim, MD, PhD

Department of Ophthalmology, Catholic University of Daegu College of Medicine, Daegu, Korea

Address reprint requests to Jae Woo Kim, MD, PhD Department of Ophthalmology, Catholic University of Daegu College of Medicine #3056-6 Daemyeung-4-dong, Nam-gu, Daegu 705-718, Korea Tel: 82-53-650-4728, Fax: 82-53-627-0133, E-mail: jwkim@cu.ac.kr

Received 5 April 2009 Accepted 9 December 2009

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

To investigate the effect of high glucose concentration on the production of reactive oxygen species in cultured human trabecular meshwork cells (HTMCs).

Methods

Primarily cultured HTMCs were exposed to low glucose (5 mM) and high glucose (22 mM) concentrations, respectively, for seven days. Cellular survival, as well as nitric oxide (NO) and hydrogen peroxide production, were assessed by measured MTT assay, Griess assay, and o-Dianisidine dihydrochloride assay, respectively. Some cells were co-exposed to N-acetyl cysteine (NAC) to assess the effect of this antioxidant.

Results

High glucose concentration increased the survival of cultured HTMCs significantly, with no effect from NAC. High glucose concentration increased the production of NO and hydrogen peroxide, which were abolished by co-exposure with NAC.

Conclusions

High glucose concentration increases the production of NO and hydrogen peroxide, which can be abolished by antioxidant in trabecular meshwork cells.

Keywords: High glucose, Hydrogen peroxide, Nitric oxide, Reactive oxygen species, Trabecular meshwork cells

References

1. Alvarado J, Murphy C, Juster R. Trabecular meshwork cellularity in primary open-angle glaucoma and nonglaucomatous normals. Ophthalmology. 1984; 91:564–79.

2. Rohen JW, LÜtjen-Drecoll E, FlÜgel C, et al. Ultrastructure of the trabecular meshwork in untreated cases of primary open-angle glaucoma. Exp Eye Res. 1993; 56:683–92.

3. Alvarado JA, Alvarado RG, Yeh RF, et al. A new insight into the cellular regulation of aqueous outflow: how trabecular meshwork endothelial cells drive a mechanism that regulates the permeability of Schlemm's canal endothelial cells. Br J Ophthalmol. 2005; 89:1500–5.

4. Schuman JS, Erickson K, Nathanson JA. Nitrovasodilator effects on intraocular pressure and ocular outflow facility in monkeys. Exp Eye Res. 1994; 58:99–105.

5. Wang RF, Podos SM. Effect of the topical application of nitro-glycerin on intraocular pressure in normal and glaucomatous monkeys. Exp Eye Res. 1995; 60:337–9.

6. Nathanson JA, McKee M. Alteration of ocular nitric oxide synthase in human glaucoma. Invest Ophthalmol Vis Sci. 1995; 36:1774–84.

7. Matsuo T. Basal nitric oxide production is enhanced by hydraulic pressure in cultured human trabecular cells. Br J Ophthalmol. 2000; 84:631–5.

8. Saccà SC, Izzotti A, Rossi P, Traverso C. Glaucomatous outflow pathway and oxidative stress. Exp Eye Res. 2007; 84:389–99.

9. Wei YH. Oxidative stress and mitochondrial DNA mutations in human aging. Proc Soc Exp Biol Med. 1998; 217:53–63.

10. Saccà SC, Pascotto A, Camicione P, et al. Oxidative DNA damage in the human trabecular meshwork: clinical correlation in patients with primary open-angle glaucoma. Arch Ophthalmol. 2005; 123:458–63.

11. Brodsky SV, Morrishow AM, Dharia N, et al. Glucose scavenging of nitric oxide. Am J Physiol Renal Physiol. 2001; 280:480–6.

12. El-Remessy AB, Abou-Mohamed G, Caldwell RW, Caldwell RB. High glucose-induced tyrosine nitration in endothelial cells: role of eNOS uncoupling and aldose reductase activation. Invest Ophthalmol Vis Sci. 2003; 44:3135–43.

13. Becker B. Diabetes mellitus and primary open-angle glaucoma. Am J Ophthalmol. 1971; 1:1–16.

14. Davies PD, Duncan G, Pynsent PB, et al. Aqueous humour glucose concentration in cataract patients and its effect on the lens. Exp Eye Res. 1984; 39:605–9.

15. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65:55–63.

16. Green LC, Wagner DA, Glogowski J, et al. Analysis of nitrate, nitrite and [15N]nitrate in biological fluids. Anal Biochem. 1982; 126:131–8.

17. Ballinger SW, Van Houten B, Jin GF, et al. Hydrogen peroxide causes significant mitochondrial DNA damage in human RPE cells. Exp Eye Res. 1999; 68:765–72.

18. Nowak D. Hydrogen peroxide release from human polymor-phonuclear leukocytes measured with horseradish peroxidase and o-dianisidine. Effect of various stimulators and cytochalasin B. Biomed Biochim Acta. 1990; 49:353–62.

19. Polansky JR, Weinreb RN, Baxter JD, Alvarado J. Human trabecular cells. I. Establishment in tissue culture and growth characteristics. Invest Ophthalmol Vis Sci. 1979; 18:1043–9.

20. Alvarado JA, Wood I, Polansky JR. Human trabecular cells. II. Growth pattern and ultrastructural characteristics. Invest Ophthalmol Vis Sci. 1982; 23:464–78.

21. Haefliger IO, Dettmann E, Liu R, et al. Potential role of nitric oxide and endothelin in the pathogenesis of glaucoma. Surv Ophthalmol. 1999; 43:S51–8.

22. Chakravarthy U, Hayes RG, Stitt AW, et al. Constitutive nitric oxide synthase expression in retinal vascular endothelial cells is suppressed by high glucose and advanced glycation end products. Diabetes. 1998; 47:945–52.

23. Moncada S, Palmer RM, Higgs EA. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev. 1991; 43:109–42.

24. Bredt DS, Snyder SH. Nitric oxide: a physiologic messenger molecule. Annu Rev Biochem. 1994; 63:175–95.

25. Brüne B, Von Knethen A, Sandau KB. Nitric oxide and its role in apoptosis. Eur J Pharmacol. 1998; 351:261–72.

26. Nathanson JA, McKee M. Identification of an extensive system of nitric oxide-producing cells in the ciliary muscle and outflow pathway of the human eye. Invest Ophthalmol Vis Sci. 1995; 36:1765–73.

27. Geyer O, Podos SM, Mittag T. Nitric oxide synthase activity in tissues of the bovine eye. Graefes Arch Clin Exp Ophthalmol. 1997; 235:786–93.

28. Meyer P, Champion C, Schlötzer-Schrehardt U, et al. Localization of nitric oxide synthase isoforms in porcine ocular tissues. Curr Eye Res. 1999; 18:375–80.

29. Weinreb RN, Bloom E, Baxter JD, et al. Detection of glucocorticoid receptors in cultured human trabecular cells. Invest Ophthalmol Vis Sci. 1981; 21:403–7.

30. Alp NJ, Channon KM. Regulation of endothelial nitric oxide synthase by tetrahydrobiopterin in vascular disease. Arterioscler Thromb Vasc Biol. 2004; 24:413–20.

31. Vasa M, Breitschopf K, Zeiher AM, Dimmeler S. Nitric oxide activates telomerase and delays endothelial cell senescence. Circ Res. 2000; 87:540–2.

32. Zhou L, Li Y, Yue BY. Oxidative stress affects cytoskeletal structure and cell-matrix interactions in cells from ocular tissue: the trabecular meshwork. J Cell Physiol. 1999; 180:182–9.

33. Kurz DJ, Decary S, Hong Y, et al. Chronic oxidative stress compromises telomere integrity and accelerates the onset of senescence in human endothelial cells. J Cell Sci. 2004; 117:2417–26.

34. von Zglinick T. Role of oxidative stress in telomere length regulation and replicative senescence. Ann N Y Acad Sci. 2000; 908:99–110.

35. Furumoto K, Inoue E, Nagao N, et al. Age-dependent telomere shortening is slowed down by enrichment of intracellular vitamin C via suppression of oxidative stress. Life Sci. 1998; 63:935–48.

36. Sato T, Roy S. Effect of high glucose on fibronectin expression and cell proliferation in trabecular meshwork cells. Invest Ophthalmol Vis Sci. 2002; 43:170–5.

37. Chen JZ, Kadlubar FF. A new clue to glaucoma pathogenesis. Am J Med. 2003; 114:697–8.

38. Wordinger RJ, Clark AF. Effects of glucocorticoids on the trabecular meshwork: towards a better understanding of glaucoma. Prog Retin Eye Res. 1999; 18:629–67.

Figure 1.

Effect of high glucose (HG) on the proliferation of human trabecular meshwork cells. HG increased cellular proliferation significantly compared to low glucose (LG) (^* p<0.05). Co-exposed 50 or 100 μM N-acetylcysteine (NAC) did not affect the HG-induced increased cellular proliferation.

Figure 2.

Effect of high glucose (HG) on the production of nitric oxide (NO) in primarily cultured human trabecular meshwork cells. HG increased NO production significantly compared to low glucose (LG) (^* p<0.05), which was abolished by co-exposed 50 or 100 μM N-acetylcysteine (NAC) (^**p<0.05).

Figure 3.

Effect of high glucose (HG) on the production of hydrogen peroxide of human trabecular meshwork cells. HG increased hydrogen peroxide production significantly compared to low glucose (LG) (^* p<0.05), which was abolished by co-exposed 50 or 100 μM N-acetylcysteine (NAC) (^** p<0.05).

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