Erkan YURTCU, Ezgi KASAPOĞLU, Feride İffet ŞAHİN

Protective effects of β-carotene and silymarin on human lymphocytes

Diyete ek olarak kullanılan beta-karoten ve silimarinin oksidatif hasara karşı koruyucu antioksidan etkileri bilinmektedir. Bu çalışmada insan lenfositlerinde L-arjinin ile oluşturulan oksidatif hasara karşı β-karoten ve silimarinin koruyucu etkilerinin varlığının değerlendirilmesi amaçlanmıştır. Çalışma kapsamına 6 sağlıklı bireyden (3 kadın, 3 erkek) alınan periferik kan örneklerinden hazırlanan lenfosit kültürleri dahil edildi. Oksidatif DNA hasarı L-arjinin ile indüklendi. β-karoten ve silimarin koruyucu etkilerinin varlığının araştırılması amacıyla kültürlere eklendi. DNA hasarının değerlendirilmesi için alkali tek hücre jel elektroforezi uygulandı. L-arjinin kültürlerde oksidatif hasarı indüklerken β-karoten ve silimarin oluşan genotoksisitenin geriye dönüşünde olumlu etki sağladı. Beta-karoten ve silimarin birlikte uygulandığında ise oluşan genotoksik hasarın, ajanların tek tek uygulanması ile elde edilenden farklı olmadığı görüldü. Verilerimiz, beta karoten ve silimarinin in vitro lenfosit kültürlerinde koruyucu etkilerini gösteren karşılaştırmalı ön çalışma sonuçlarını içermektedir.

Beta-karoten ve silimarinin insan lenfositleri üzerindeki koruyucu etkisi

Beta-carotene and silymarin have antioxidant properties against oxidative damage and are used as dietary supplements. The aim of this study was to assess the protective effects of β-carotene and silymarin on healthy human lymphocytes exposed to L-arginine-induced oxidative damage. Study samples were lymphocyte cultures set up from venous blood obtained from 6 healthy individuals (3 males and 3 females). Oxidative DNA damage was induced by L-arginine. β-Carotene and silymarin were added to the cultures as antioxidants to observe their protective effects, if any. Alkaline single-cell gel electrophoresis (SCGE) was used to evaluate DNA damage. While L-arginine generated genotoxic damage in the lymphocyte cultures, both β-carotene and silymarin had positive effects on the reversal of the induced genotoxicity. When β-carotene was applied in combination with silymarin, there was no significant change in mean tail moments in comparison to the use of β-carotene or silymarin alone. The data presented here provide a comparative, preliminary insight into the protective role of both β-carotene and silymarin on L-arginine-induced genotoxicity in in vitro lymphocyte cultures.

PDF

___

1. Zhang P, Omaye ST. DNA strand breakage and oxygen tension: effects of beta-carotene, alpha-tocopherol and ascorbic acid. Food Chem Toxicol 39: 239-246, 2001.
2. Sies H, Stahl W. Vitamins E and C, beta-carotene, and other carotenoids as antioxidants. Am J Clin Nutr 62: 1315S-1321S, 1995.
3. Ray SD, Lam TS, Rotollo JA et al. Oxidative stress is the master operator of drug and chemically-induced programmed and unprogrammed cell death: Implications of natural antioxidants in vivo. Biofactors 21: 223-232, 2004.
4. Orhan İ, Özçelik B, Şener B. Evaluation of antibacterial, antifungal, antiviral, and antioxidant potentials of some edible oils and their fatty acid profi les. Turk J Biol 35: 251-258, 2011.
5. Ansari MA, Keller JN, Scheff SW. Protective effect of Pycnogenol in human neuroblastoma SH-SY5Y cells following acrolein-induced cytotoxicity. Free Radic Biol Med 45: 1510- 1519, 2008.
6. Krinsky NI, Johnson EJ. Carotenoid actions and their relation to health and disease. Mol Aspects Med 26: 459-516, 2005.
7. Comelli MC, Mengs U, Schneider C et al. Toward the definition of the mechanism of action of silymarin: activities related to cellular protection from toxic damage induced by chemotherapy. Integr Cancer Ther 6: 120-129, 2007.
8. Post-White J, Ladas EJ, Kelly KM. Advances in the use of milk thistle (Silybum marianum). Integr Cancer Ther 6: 104-109, 2007.
9. Ladas EJ, Kelly KM. Milk thistle: is there a role for its use as an adjunct therapy in patients with cancer? J Altern Complement Med 9: 411-416, 2003.
10. Cemek M, Enginar H, Karaca T et al. In vivo radioprotective effects of Nigella sativa L oil and reduced glutathione against irradiation-induced oxidative injury and number of peripheral blood lymphocytes in rats. Photochem Photobiol 82: 1691- 1696, 2006.
11. Erden CD, Ekmekci A, Sahin FI et al. L-arginine and mitomycin C-induced nitric oxide release and apoptosis in human lymphocytes. Cell Biol Int 27: 337-340, 2003.
12. Bal F, Sahin FI, Yirmibes M et al. The in vitro effect of betacarotene and mitomycin C on SCE frequency in Down’s syndrome lymphocyte cultures. Tohoku J Exp Med 184: 295- 300, 1998.
13. Yilmaz Z, Karabay G, Oktem M et al. The apoptotic eff ects of mitomycin C on human endometrial cell cultures and reversal of its effects by beta-carotene and folic acid. Acta Physiol Hung 93: 41-51, 2006.
14. Micozzi MS, Brown ED, Edwards BC et al. Plasma carotenoid response to chronic intake of selected foods and β-carotene supplements in men. Am J Clin Nutr 55: 1120-1125, 1992.
15. Wen Z, Dumas TE, Schrieber SJ et al. Pharmacokinetics and metabolic profi le of free, conjugated, and total silymarin fl avonolignans in human plasma aft er oral administration of milk thistle extract. Drug Metab Dispos 36: 65-72, 2008.
16. O’Brien NM, Woods JA, Aherne SA et al. Cytotoxicity, genotoxicity and oxidative reactions in cell-culture models: modulatory eff ects of phytochemicals. Biochem Soc Trans 28: 22-26, 2000.
17. Stanner SA, Hughes J, Kelly CN et al. A review of the epidemiological evidence for the ‘antioxidant hypothesis’. Public Health Nutr 7: 407-422, 2004.
18. Duthie SJ, Ma A, Ross MA et al. Antioxidant supplementation decreases oxidative DNA damage in human lymphocytes. Cancer Res 56: 1291-1295, 1996.
19. Panayiotidis M, Collins AR. Ex vivo assessment of lymphocyte antioxidant status using the comet assay. Free Radic Res 27: 533-537, 1997.
20. Woods JA, Bilton RF, Young AJ. Beta-carotene enhances hydrogen peroxide-induced DNA damage in human hepatocellular HepG2 cells. FEBS Lett 449: 255-258, 1999.
21. Martin KR, Failla ML, Smith JC Jr. Beta-carotene and lutein protect HepG2 human liver cells against oxidant-induced damage. J Nutr 126: 2098-2106, 1996.
22. Konopacka M, Rzeszowska-Wolny J. Antioxidant vitamins C, E and beta-carotene reduce DNA damage before as well as after gamma-ray irradiation of human lymphocytes in vitro. Mutat Res 491: 1-7, 2001.
23. Wellington K, Jarvis B. Silymarin: a review of its clinical properties in the management of hepatic disorders. Bio Drugs 15: 465-489, 2001.
24. Alidoost F, Gharagozloo M, Bagherpour B et al. Effects of silymarin on the proliferation and glutathione levels of peripheral blood mononuclear cells from beta-thalassemia major patients. Int Immunopharmacol 6: 1305-1310, 2006.
25. Liebler DC. Antioxidant reactions of carotenoids. Ann NY Acad Sci 691: 20-31, 1993.
26. Lowe GM, Booth LA, Young AJ et al. Lycopene and betacarotene protect against oxidative damage in HT29 cells at low concentrations but rapidly lose this capacity at higher doses. Free Radic Res 30: 141-151, 1999.
27. Buettner GR. The pecking order of free radicals and antioxidants: lipid peroxidation, alpha-tocopherol, and ascorbate. Arch Biochem Biophys 300: 535-543, 1993.
28. Kaya Gİ, Somer NÜ, Konyalıoğlu S et al. Antioxidant and antibacterial activities of Ranunculus marginatus var. trachycarpus and R. sprunerianus. Turk J Biol 34: 139-146, 2010.