Korean J Ophthalmol.  2008 Sep;22(3):183-186. 10.3341/kjo.2008.22.3.183.

Protective Effects of Epigallocatechin Gallate After UV Irradiation of Cultured Human Lens Epithelial Cells

• Heo J
• Lee BR
• Koh JW

• KMID: 1070755
• DOI: http://doi.org/10.3341/kjo.2008.22.3.183

Abstract

PURPOSE: To evaluate the protective effects of epigallocatechin gallate (EGCG) against UV irradiation of cultured human lens epithelial cells. METHODS: We irradiated cultured human lens epithelial cells with a 30-second pulse from a UV lamp with an irradiance of 0.6 mW/cm2. Five minutes and 1 hour after UV irradiation, we administered 0, 5, 10, 15, 25, 50, or 100 uM EGCG. The cell number was measured with a microscopic counting chamber and cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: Compared to untreated cells, the total number of cultured human lens epithelial cells was markedly higher after UV irradiation. In a dose-dependent manner, viability was also higher in EGCG-treated cells. CONCLUSIONS: EGCG increased the cell count and cell viability after UV irradiation of cultured human lens epithelial cells, indicating that EGCG can protect lens epithelium against UV damage.

Keyword

Cell viability; Cultured human lens epithelial cells; Epigallocatechin gallate (EGCG); 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay; UV irradiation

MeSH Terms

Antioxidants/*pharmacology
Catechin/*analogs & derivatives/pharmacology
Cell Count
Cell Survival/drug effects
Cells, Cultured
Coloring Agents/diagnostic use
Dose-Response Relationship, Drug
Epithelial Cells/radiation effects
Humans
Lens, Crystalline/cytology/*radiation effects
Radiation Injuries/*prevention & control
Radiation-Protective Agents/*pharmacology
Tetrazolium Salts/diagnostic use
Thiazoles/diagnostic use
*Ultraviolet Rays

Figure

• 

  Fig. 1 Lens epithelial cell loss after UV irradiation. (A) Control without UV irradiation. (B) After UV irradiation without administration of Epigallocatechin gallate (EGCG). (C) 5 minutes after UV irradiation with administration of 100 µM EGCG. (D) 1 hour after UV irradiation with administration of 100 µM EGCG. The photographs were taken by inverted microscope. (×200)

• 

  Fig. 2 Lens epithelial cell count after UV irradiation. Cell count of cultured human lens epithelial cells after UV irradiation was markedly increased with Epigallocatechin gallate (EGCG) administrationgroup, compared with non- administration group. Control was group without UV irradiation. There was no significant relationship between the time to administration EGCG and the lossof cells (P=0.719). (A) 5 minutes after UV irradiation, (B) 1 hour after UV irradiation.

• 

  Fig. 3 Lens epithelial cell viability after UV irradiation. Cell viability of cultured human lens epithelial cells after UV irradiation was markedly increased with Epigallocatechin gallate (EGCG) administration group and was increased dose-dependent way. Basis (0) was group with UV irradiation without administration of EGCG. There was no significance between the time to administration of EGCG and the survival rate of cells (P=0.129). (A) 5 minutes after UV irradiation, (B) 1 hour after UV irradiation.

Reference

1. McCarty CA, Taylor HR. Recent developments in vision research: light damage in cataract. Invest Ophthalmol Vis Sci. 1996. 37:1720–1723.

2. Norren DV, Vos JJ. Spectral transmission of the human ocular media. Vision Res. 1974. 14:1237–1244.

3. Margrain TH, Boulton M, Marshall J, et al. Do blue light filters confer protection against age-related macular degeneration. Prog Retin Eye Res. 2004. 23:523–531.

4. Worgul BV, Merriam GR Jr, Medvedovsky C. Cortical cataract development-an expression of primary damage to the lens epithelium. Lens Eye Toxicity Res. 1989. 6:559–571.

5. Taylor HR, West SK, Rosenthal FS, et al. Effect of ultraviolet radiation in cataract formation. N Engl J Med. 1998. 319:1429–1433.

6. Zigman S, Vaughan T. Near-ultraviolet light effects on the lenses and retinas of mice. Invest Ophthalmol Vis Sci. 1974. 13:462–465.

7. Spector A, Garner WH. Hydrogen peroxide and human cataract. Exp Eye Res. 1981. 33:673–681.

8. Ramachandran S, Morris SM, Devamanoharan P, et al. Radio-isotopic determination of hydrogen peroxide in aqueous humor and urine. Exp Eye Res. 1991. 53:503–506.

9. Middleton E, Kandaswami C, Theoharides TC. The effects of plant flavonoids on mammalian cells: Implicationsfor inflammation, heart disease and cancer. Pharmacol Rev. 2000. 52:673–751.

10. Bettuzzi S, Brausi M, Rizzi F, et al. Chemoprevention of human prostate cancer by oral administration of green tea catechins in volunteers with high-grade prostate intraepithelial neoplasia: a preliminary report from a one-year proof-of-principle study. Cancer Res. 2006. 66:1234–1240.

11. Demeule M, Michaud-Levesque J, Annabi B, et al. Green tea catechins as novel antitumor and antiangiogenic compounds. Curr Med Chem Anticancer Agents. 2002. 2:441–463.

12. Nagle DG, Ferreira D, Zhou YD. Epigallocatechin-3-gallate (EGCG): Chemical and biomedical perspectives. Phytochemistry. 2006. 67:1849–1855.

13. Buttemeyer R, Philipp AW, Schlenzka L, et al. Epigallocatechin gallate can significantly decrease free oxygen radicals in the reperfusion injury in vivo. Transplant Proc. 2003. 35:3116–3120.

14. Morley N, Clifford T, Salter LA, et al. The green tea polyphenol (-)-epigallocatechin gallate and green tea can protect human cellular DNA from ultraviolet and visible radiation-induced damage. Photodermatol Photoimmunol Photomed. 2005. 21:15–22.

15. Nagai K, Jiang MH, Hada J. (-)-Epigallocatechin gallate protects against NO stress-induced neuronal damage after ischemia by acting as an anti-oxidant. Brain Res. 2002. 956:319–322.

16. Xie D, Liu G, Zhu G, et al. (-)-Epigallocatechin-3-gallate protects cultured spiral ganglion cells from H2O2-induced oxidizing damage. Acta Otolaryngol. 2004. 124:464–470.

17. Mandel S, Weinreb O, Amit T, Youdim MB. Cell signaling pathways in the neuroprotective actions of the green tea polyphenol (-)-epigallocatechin-3-gallate: implications for neurodegenerative diseases. J Neurochem. 2004. 88:1555–1569.

18. Skrzydlewska E, Ostrowska J, Farbiszewski R, Michalak K. Protective effect of green tea against lipid peroxidation in the rat liver, blood serum and the brain. Phytomedicine. 2002. 9:232–238.

19. Yokozawa T, Nakagawa T, Kitani K. Antioxidative activity of green tea polyphenol in cholesterol-fed rats. J Agric Food Chem. 2002. 50:3549–3552.

20. Negishi H, Xu JW, Ikeda K, et al. Black and green tea polyphenols attenuate blood pressure increases in stroke-prone spontaneously hypertensive rats. J Nutr. 2004. 134:38–42.

21. Sutherland BA, Rahman RM, Appleton I. Mechanisms of action of green tea catechins, with a focus on ischemiainduced neurodegeneration. J Nutr Biochem. 2006. 17:291–306.