Tannik Asitin İnsan Lenfositlerinde DNA Hasarı Üzerine Etkisi

Year 2014, Issue: 2, 133 - 146, 01.06.2014

Dilek Tokaç Sevtap Aydın Arif Ahmet Başaran Nurşen Başaran

Abstract

Doğal ürünlerde bulunan fenolik maddeler reaktif oksijen türevlerinin zararlı etkilerine karşı organizmayı korumada önemli bir role sahiptir. Suda çözünen polifenolik tannik asit TA bitkisel çaylar, bira, ceviz, fındık ve üzümsü meyveler gibi pek çok bitkisel içecek ve gıdalarda bulunmaktadır. Tanenlerin kan pıhtılaşmasını hızlandırdığı, kan basıncını azalttığı, serum lipid düzeyini düşürdüğü, karaciğer nekrozuna neden olduğu ve immün yanıtları düzenlediği gibi bazı fizyolojik etkileri bildirilmektedir. TA konsantrasyonuna ve yapısında bulunan hidroksil grupların sayı ve bulunduğu yere göre antioksidan veya prooksidan olarak rol almaktadır. Ancak, TA’ca zengin gıdaların tüketiminin özofagus kanseri gibi belli kanser sıklıkları ile ilişkili olması nedeniyle TA’nın karsinojenik olabileceği de ileri sürülmektedir. Bu çalışmada TA’nın antioksidan kapasitesi troloks eşdeğer antioksidan kapasite deneyi ile ve insan lenfositlerinde H2O2 ile indüklenen DNA hasarı üzerine TA’nın etkisi standart comet yöntemi ve formamidoprimidin-DNA-glikosilaz Fpg ile adapte edilmiş comet yöntemi ile araştırılmıştır. Sonuçlarımız 0.5 μM konsantrasyon üzerinde TA’nın anlamlı olarak antioksidan kapasiteye sahip olduğunu göstermiştir. Çalışılan tüm konsantrasyonlarında 0.1-100 M TA’nın tek başına DNA hasarına neden olmadığı ve hatta H2O2 ile indüklenen oksidatif DNA hasarını anlamlı olarak azalttığı saptanmıştır. Ayrıca 0.1-100 M konsantrasyon aralığında tek başına TA’nın artmış okside pürin baz düzeylerinin göstergesi olan Fpg duyarlı bölgeleri indüklemediği, ancak TA’nın çalışılan tüm konsantrasyonlarında doz bağımlı olarak H2O2 ile indüklenen Fpg duyarlı bölgeleri anlamlı olarak azalttığı saptanmıştır. Sonuç olarak, TA’nın insan lenfositlerinde oksidatif DNA hasarı üzerine koruyucu bir etkiye sahip olduğu in vitro olarak görülmektedir. Diyetsel destek olarak uygun düzeylerde tannik asit doğal bir antioksidan olarak kullanılabilir, fakat tannik asitin insan sağlığı üzerine potansiyel etkilerini aydınlatmak için daha ileri in vivo ve in vitro çalışmalara gerek vardır.

Keywords

Tannik asit, Hidrojen peroksit, Tek hücre jel elektroforez, Comet yöntemi, TEAK Troloks eşdeğer antioksidan kapasite yöntemi

The Effect of Tannic Acid on DNA Damage in Human Lymphocytes

Abstract

Phenolic substances in natural products play an important role in protecting the organism against harmful effects of reactive oxygen species. Tannic acid TA , water-soluble polyphenol, is present in many plant beverages and foods, such as herbal teas, beer, walnut, hazelnut, and berries. Tannins have been reported to exert some physiological effects, such as to accelerate blood clotting, reduce blood pressure, decrease the serum lipid level, cause liver necrosis, and modulate immunoresponses. It acts as an antioxidant or a prooxidant, depending on the concentrations of TA and the number and position of hydroxyl groups on TA. It has been suggested that tannins might be carcinogenic, as the incidences of certain cancers, such as esophageal cancer, are related to consumption of tannins-rich foods. In the present study, the antioxidant capacity of TA was determined by the trolox equivalent antioxidant capacity assay and the effect of TA on DNA damage induced by H2O2 in human lymphocytes were investigated by the standard comet assay and the formamidopyrimidine-DNA-glycoslase Fpg modified comet assay. Our study showed that above the concentrations of 0.5 μM TA showed significant antioxidant capacity. It was determined that at all the concentrations studied 0.1-100 M , TA alone did not cause DNA damage and even significantly reduced oxidative DNA damage induced by H2O2. It was also determined that within the concentration of 0.1-100 M TA alone did not induce Fpg sensitive sites indicating the increased oxidized purine base levels, but TA treatment at all the concentrations studied significantly reduced the Fpg sensitive sites induced by H2O2 in a dose dependent. In conclusion, it seems that TA exerts a protective effect on oxidative DNA damage in human lymphocytes in vitro. In appropriate levels as a dietary supplement, TA may be used as a natural antioxidant however it is necessary to perform the further in vivo and in vitro studies to clarify the potential effects of TA on human health.

Keywords

Tannic acid, Hydrogen peroxide, Single cell gel electrophoresis, Comet assay, TEAC Trolox equivalent antioxidant capacity assay

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