Bazı bitkisel antioksidanların mutajenik ve antimutajenik etkilerinin kısa süreli genotoksisite test yöntemleri ile belirlenmesi
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Date
2010-06-11
Authors
Zorlu, Tolga
Journal Title
Journal ISSN
Volume Title
Publisher
Uludağ Üniversitesi
Abstract
Bu çalışmada fiziksel bir mutajen ajan olan iyonize radyasyonun genotoksik etkisi ve fenolik bir asit olan kinik asidin radyoprotektif etkisi çalışılmıştır. Bu amaçla insan periferal kan lenfositlerinde in vitro COMET testi uygulanmıştır.Bireylerden heparinize olarak alınan periferal kandan lenfosit izolasyonu yapılmıştır. İzole edilen lenfositler kültüre edilmiş ve kültür sonunda hücreler 1 ve 2 Gy radyasyona maruz bırakılmıştır. Radyasyon maruziyetinden 30 dakika önce hücreler 0,5, 1, 2, 4 ve 8 µg/ml kinik asit dozları ile inkübe edilmiştir. Bu işlemlerde sonra in vitro COMET testi gerçekleştirilmiştir. COMET testi sonuçlarına göre radyasyon doz grupları kendi aralarında ve kontrol grubu karşılaştırıldığında Genetik Hasar İndeksi ortalama değerleri arasında anlamlı bir farklılık bulunmuştur (p<0,001). Yine kinik asit dozları ve kontrol grubu arasında yapılan istatistiksel hesaplamalar sonucunda, bu dozların Genetik Hasar İndeksi bakımından kontrol grubu ve kendi aralarında herhangi bir anlamlı farklılık gözlenmemiştir (p>0,05).Kinik asidin 0,5 µg/ml, 1 µg/ml, 2 µg/ml, 4 µg/ml ve 8 µg/ml dozları, radyasyonun 1 Gy ve 2 Gy radyasyon dozları ile kombine edildiğinde, kombine dozlar ve tek başına radyasyon dozları (1 Gy ve 2 Gy) karşılaştırıldığında aralarında anlamlı farklılık bulunmuştur (p<0,001). 1 Gy ve 0,5 µg/ml kombine dozu ile 1 Gy tek radyasyon grubu arasında istatistiksel olarak anlamlı bir farklılık bulunamamıştır. (p>0,05). Aynı şekilde 2 Gy + 0,5 µg/ml kombine dozu ile 2 Gy tek radyasyon arasında istatistiksel olarak anlamlı bir farklılık yoktur (p>0,05). Kinik asidin 4 µg/ml'lık dozu, 1 ve 2 Gy radyasyon dozlarının indüklediği DNA hasarını en etkili şekilde azaltan doz grubu olduğu belirlenmiştir.Sonuç olarak radyasyonun 1 ve 2 Gy' lik dozları COMET testine göre genotoksik etki göstermiştir. Fenolik hidroksil gruplar taşıyan kinik asit ise, bu radyasyonun indüklediği genetik hasarı azaltmıştır. COMET testi sonuçlarına göre kinik asitin radyoprotektif etkisinin olduğu ortaya konmuştur.
In our study, genotoxic effects of ionizing radiation which is a physical mutagen and radioprotective effects of quinic acid are evaluated. For this purpose in vitro COMET assay was carried out in human peripheral blood lymphocytes.Blood samples are drawn into tubes with heparin as an anticoagulant and lymphocytes are isolated. Isolated lymphocytes were cultured. After 24 hour following culture initiation, the lymphocytes were incubated with 0,5, 1, 2, 4 and 8 µg/ml doses of quinic acid. Thirty minutes after quinic acid treatment, all lymphocyte culture petri dishes excluding control lymphocytes were exposed to ionizing radiation. Once viabilities were ensured, more than 80%, the lymphocytes were subjected to the COMET assay after irradiation. According to COMET assay results, when radiation dose groups were compared among themselves and with control groups, significant differences were detected between Genetic Damage Index average values of dose groups (p<0,001). In quinic acid alone treated lymphocytes, no significant differences were detected when compared to control group. (p>0,05).When 0,5, 1, 2, 4 and 8 µg/ml doses of quinic acid were combined with radiation doses, significant differences were detected when compared among the quinic acid combined dose groups themselves and radiation dose groups (p<0,001). In 1 Gy + 0,5 µg/ml quinic acid study group, no significant differences were found in the Genetic Damage Index average values when compared to 1 Gy radiation dose group (p>0,05) . Also no significant differences detected in 2 Gy + 0,5 µg/ml quinic acid group when compared with 2 Gy study group (p>0,05). From the above-mentioned results, 4 µg/ml of quinic acid was selected as the effective concentration which effectively decreased the DNA damage that induced by 1 and 2 Gy doses of ionizing radiation.In present study demonstrates that 1 and 2 Gy doses of radiation showed genotoxic effects in human peripheral blood lymphocytes. İonizing radiation induced genetic damage is decreased by quinic acid which comprehends phenyl hydroxyl groups. The current study demonstrates that quinic acid is effective in protecting lymphocytes against radiation-induced toxicity.
In our study, genotoxic effects of ionizing radiation which is a physical mutagen and radioprotective effects of quinic acid are evaluated. For this purpose in vitro COMET assay was carried out in human peripheral blood lymphocytes.Blood samples are drawn into tubes with heparin as an anticoagulant and lymphocytes are isolated. Isolated lymphocytes were cultured. After 24 hour following culture initiation, the lymphocytes were incubated with 0,5, 1, 2, 4 and 8 µg/ml doses of quinic acid. Thirty minutes after quinic acid treatment, all lymphocyte culture petri dishes excluding control lymphocytes were exposed to ionizing radiation. Once viabilities were ensured, more than 80%, the lymphocytes were subjected to the COMET assay after irradiation. According to COMET assay results, when radiation dose groups were compared among themselves and with control groups, significant differences were detected between Genetic Damage Index average values of dose groups (p<0,001). In quinic acid alone treated lymphocytes, no significant differences were detected when compared to control group. (p>0,05).When 0,5, 1, 2, 4 and 8 µg/ml doses of quinic acid were combined with radiation doses, significant differences were detected when compared among the quinic acid combined dose groups themselves and radiation dose groups (p<0,001). In 1 Gy + 0,5 µg/ml quinic acid study group, no significant differences were found in the Genetic Damage Index average values when compared to 1 Gy radiation dose group (p>0,05) . Also no significant differences detected in 2 Gy + 0,5 µg/ml quinic acid group when compared with 2 Gy study group (p>0,05). From the above-mentioned results, 4 µg/ml of quinic acid was selected as the effective concentration which effectively decreased the DNA damage that induced by 1 and 2 Gy doses of ionizing radiation.In present study demonstrates that 1 and 2 Gy doses of radiation showed genotoxic effects in human peripheral blood lymphocytes. İonizing radiation induced genetic damage is decreased by quinic acid which comprehends phenyl hydroxyl groups. The current study demonstrates that quinic acid is effective in protecting lymphocytes against radiation-induced toxicity.
Description
Keywords
İyonize radyasyon, Kinik asit, Comet testi, Antioksidan, Genotoksisite, Radyoprotektörler, Ionizing Radiation, Quinic acid, COMET assay, Antioxidant, Genotoxicity, Radioprotection
Citation
Zorlu, T. (2010). Bazı bitkisel antioksidanların mutajenik ve antimutajenik etkilerinin kısa süreli genotoksisite test yöntemleri ile belirlenmesi. Yayınlanmamış yüksek lisans tezi. Uludağ Üniversitesi Fen Bilimleri Enstitüsü.