Casuarinin Protects Cultured MDCK Cells from Hydrogen Peroxide-Induced Oxidative Stress and DNA Oxidative Damage

Abstract

Casuarinin has been shown to be an antioxidant in acellular experiments. This study was designed to assess the ability of casuarinin, extracted from Terminalia arjuna, to protect cultured Madin-Darby canine kidney (MDCK) cells against H₂O₂-mediated oxidative stress. A comparison with trolox, a hydrosoluble vitamin E analogue was performed. MDCK cells were pretreated with casuarinin or trolox for 1 h, then exposed to H₂O₂. After incubation with 0.8 mM H₂O₂ for 1 h, casuarinin caused a decrease in intracellular peroxide production as shown by dichlorofluorescein (DCF) fluorescence in a concentration-dependent manner. After 3 h exposure to 8 mM H₂O₂, the percentage of intracellular glutathione (GSH)-negative cells was reduced in the casuarinin-treated group. Addition of 32mM H₂O₂ to MDCK cells for 3 h induced an increase in the percentage of cells containing 8-oxoguanine but the level of such cells declined in casuarinin-treated cells. These results show that casuarinin is more effective against H₂O₂-induced oxidative damage than trolox. The data suggest that casuarinin attenuates H₂O₂-induced oxidative stress, decreases DNA oxidative damage and prevents the depletion of intracellular GSH in MDCK cells.

Key words

Casuarinin - Terminalia arjuna - oxidative stress - hydrogen peroxide - glutathione - DNA oxidative damage

References

• 1 Gupta R, Singhal S, Goyle A, Sharma V N. Antioxidant and hypocholesterolaemic effects of Terminalia arjuna tree-bark powder: a randomised placebo-controlled trial.  J Assoc Physicians India. 2001;  49 231-5
• 2 Shaila H P, Udupa S L, Udupa A L. Hypolipidemic activity of three indigenous drugs in experimentally induced atherosclerosis.  Int J Cardiol. 1998;  67 119-24
• 3 Kaur S, Grover I S, Kumar S. Antimutagenic potential of extracts isolated from Terminalia arjuna .  J Environ Pathol Toxicol Oncol. 2001;  20 9-14
• 4 Perumal Samy R, Ignacimuthu S, Sen A. Screening of 34 Indian medicinal plants for antibacterial properties.  J Ethnopharmacol. 1998;  62 173-82
• 5 Yang L L, Lee C Y, Yen K Y. Induction of apoptosis by hydrolysable tannins from Eugenia jambos L. on human leukemia cells.  Cancer Lett. 2000;  157 65-75
• 6 Kashiwada Y, Nonaka G, Nishioka I, Chang J J, Lee K H. Antitumor agents, 129. Tannins and related compounds as selective cytotoxic agents.  J Nat Prod. 1992;  55 1033-43
• 7 Satomi H, Umemura K, Ueno A, Hatano T, Okuda T, Noro T. Carbonic anhydrase inhibitors from the pericarps of Punica granatum L.  Biol Pharm Bull. 1993;  16 787-90
• 8 Maclin L J, Bendich A. Free radical tissue damage: protective role of antioxidant nutrients.  FASEB J. 1987;  1 441-5
• 9 Ishii R, Saito K, Horie M, Shibano T, Kitanaka S, Amano F. Inhibitory effects of hydrolysable tannins from Melastoma dodecandrum Lour. on nitric oxide production by murine macrophage-like cell line, RAW264.7, activated with lipopolysaccharide and interferon-gamma.  Biol Pharm Bull. 1999;  22 647-53
• 10 Ames B N, Shigenaga M K, Hagen T M. Oxidants, antioxidants, and the degenerative diseases of aging.  Proc Natl Acad Sci USA. 1993;  90 7915-22
• 11 Halliwell B, Aruoma O I. DNA damage by oxygen-derived species.  FEBS Lett. 1991;  281 9-19
• 12 Lin T C, Ma Y T, Hsu F L. Tannins from the bark of Terminalia arjuna.  Chin Pharm J. 1996;  48 25-35
• 13 LeBel C P, Ischiopoulos H, Bondy S C. Evaluation of the probe 2′,7′-dichlorofluorescin as indicator of reactive oxygen species formation and oxidative stress.  Chem Res Toxicol. 1992;  5 227-31
• 14 Chang W H, Chen C H, Lu F J. Different effects of baicalein, baicalin and wogonin on mitochondrial function, glutathione content and cell cycle progression in human hepatoma cell lines.  Planta Medica. 2002;  68 128-32
• 15 Fernandez-Checa J C, Kaplowitz N, Garcia-Ruiz C, Colell A. Mitochondrial glutathione: importance and transport.  Semin Liver Dis. 1998;  18 389-401
• 16 De Zwart L L, Meerman J H, Commandeur J N, Vermeulen N P. Biomarkers of free radical damage applications in experimental animals and in humans.  Free Radical Biol Med. 1999;  26 202-26
• 17 Halliwell B, Gutteridge J MC, Cross C E. Free radicals, antioxidants, and human disease: where are we now?.  J Lab Clin Med. 1992;  119 598-620
• 18 Winterbourn C C, Metodiewa D. The reaction of superoxide with reduced glutathione.  Arch Biochem Biophys. 1994;  314 284-90
• 19 Tsai C H, Chern C L, Liu T Z. Antioxidant action of glutathione: Its interaction with superoxide anion and hydroxyl radical.  J Biomed Lab Sci (Taiwan). 2000;  12 107-11
• 20 Chandre J, Samali A, Orrenius S. Triggering and modulation of apoptosis by oxidative stress.  Free Radical Biol Med. 2000;  29 323-33