İnsan Lenfositlerinde Likopenin Oksidatif DNA Hasarı Üzerine Etkisi

Year 2013, Issue: 2, 163 - 178, 01.06.2013

Dilek Tokaç Sevtap Aydın Arif Ahmet Başaran Nurşen Başaran

Abstract

Likopen domates, işlenmiş domates ürünleri ve diğer meyvelerde yaygın olarak bulunan ve insan diyetinde önemli rol oynayan bir karotenoiddir. Bir antioksidan olarak rol oynayabildiği ve oksidatif strese karşı hücresel makromolekülleri koruyabildiği gösterilmektedir. Likopen içeren gıdaların diyetsel alımlarının reaktif oksijen radikalleri tarafından başlatıldığı düşünülen ateroskleroz, katarakt, yaşlanma ile ilişkili makula dejenerasyonu, multiple skleroz ve kanser gibi hastalıkların inhibisyonuna katkı sağladığı tahmin edilmektedir. Bu çalışma H2O2 ile indüklenen oksidatif DNA hasarına karşı likopenin etkisini incelemek üzere gerçekleştirilmiştir ve DNA hasarı tek hücre jel elektroforez comet yöntemi ile değerlendirilmiştir. İnsan lenfositlerinde H2O2 ile indüklenen okside pürin baz düzeylerini incelemek üzere formamidoprimidin-DNA-glikozilaz Fpg enzimi kullanılmıştır. Troloks eşdeğer antioksidan kapasite TEAK yöntemi ile likopenin antioksidan kapasitesi belirlendi. 10 mM üzeri çalışılan tüm konsantrasyonlarda likopenin tek başına DNA hasarını indüklediği görülmüştür. Ancak insan lenfositlerinde 0.05 mM H2O2 ile indüklenen DNA tek zincir kırıklarını 0.5-12 mM konsantrasyonlarda likopen anlamlı olarak azaltmıştır. 10mM üzeri konsantrasyonlarda tek başına likopenin artmış okside pürin düzeylerin göstergesi olan Fpg duyarlı bölgeleri indüklediği görülmektedir. Sonuç olarak, likopen in vitro insan lenfositlerinde hem antioksidan hem de oksidan özellik göstermektedir. Diyetsel destek olarak uygun düzeylerde likopen doğal bir antioksidan olarak kullanılabilir, fakat sonuçlarımız sadece insane lenfositleri ile yapılmış in vitro deneylerle alakalıdır. Likopenin mutajenik/antimutajenik potansiyelini aydınlatmak için ayrıca hayvan ve insan çalışmaları da gerçekleştirilmelidir.

Keywords

Likopen, hidrojen peroksit, tek hücre jel elektroforez, comet yöntemi, TEAK Troloks eşdeğer antioksidan kapasite yöntemi

The Effect of Lycopene on Oxidative DNA Damage in Human Lymphocytes

Abstract

Lycopene, a carotenoid compound widely present in tomatoes, processed tomato products and other fruits, plays an important role in human diet. It has been shown that lycopene could act as an antioxidant and protect cellular macromolecules against oxidative stress. Dietary intake of food containing lycopene has been estimated to contribute to the inhibition of the diseases believed to be initiated by reactive oxygen species including atherosclerosis, cataract, age-related macular degeneration, multiple sclerosis, and cancer. The present study was carried out to investigate the modulating effects of lycopene against the oxidative DNA damage induced by H2O2 and the DNA damage was evaluated by single cell gel electrophoresis comet assay . Formamidopyrimidine-DNA-glycoslase Fpg enzyme was used to examine the levels of oxidized purine bases induced by H2O2 in human lymphocytes. The antioxidant capacity of lycopene was also analyzed by trolox equivalent antioxidant capacity TEAC assay. At all concentrations studied above 10 mM lycopene alone seemed to induce DNA damage. However at the concentrations of 0.5-12 mM lycopene significantly reduced DNA single strand breakages induced by 0.05 mM H2O2 in human lymphocytes. At the concentrations above 10mM lycopene alone seemed to induce Fpg sensitive sites indicating the increased oxidized purine base levels. In conclusion, it seems that lycopene exerts both oxidant and antioxidant properties in human lymphocytes in vitro. In appropriate levels as a dietary supplement, lycopene can be used as a natural antioxidant but our results concern only in vitro experiments with human lymphocytes. Additional animal and human studies should be performed in order to clarify the mutagenic/antimutagenic potential of lycopene.

Keywords

Lycopene, hydrogen peroxide, single cell gel electrophoresis, comet assay, TEAC trolox equivalent antioxidant capacity assay

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