Cinnamic (sinnamik) asidin radyasyondan koruyucu etkisinin in vitro kısa süreli genotoksisite test yöntemleri ile araştırılması
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Date
2013
Authors
Tüzün, Ece
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Publisher
Uludağ Üniversitesi
Abstract
Bu çalışmada fiziksel bir mutajen ajan olan iyonize radyasyonun genotoksik etkisi ve fenolik bir asit olan sinnamik asidin radyoprotektif etkisi çalısılmıştır. Bu amaçla insan periferal kan lenfositlerinde in vitro comet ve mikrnonukleus 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 1, 2, 4 g/ml sinnamik asit dozları ile inkübe edilmiştir. Bu işlemlerde sonra in vitrocomet ve mikronukleus testleri gerçekleştirilmiştir. COMET testi sonuçlarına göre genetik hasar indeksi ve hasarlı hücre değerlerinden yararlanılarak karşılaştırmalar yapılmıştır. Genetik hasar indeksleri açısından 1 Gy.R. ile 1, 2, 4 g/ml sinnamik asit kombine dozları ve 2 Gy.R. ile 1, 2, 4 g/ml sinnamik asit kombine dozlarıyla yapılan ikili karşılaştırmalar açısından istatiksel olarak bir anlamlılık bulunmuştur (p<0,001). COMET testinden yararalanarak % Hasarlı Hücre değerleri ile de 1 Gy.R. ile1, 2, 4 g/ml sinnamik asit kombine dozları ve 2 Gy.R. ile 1, 2, 4 g/ml sinnamik asit kombine dozlarıyla da karşılaştırmalar yapılmış bir anlamlılık burada da bulunmuştur (p<0,005). Mikronukleus testinden yaralanılarak MN frekansı ve Nükleer Bölünme İndeksi (NBİ) ortalama değerlerine göre gerekli istatiksel test yöntemleri kullanılarak karşılaştırmalar yapılmıştır. NBİ değerleri açısından bir anlamlık bulunmamıştır (p>0,05). MN frekansı açısından ise 1 GyR. ile1, 2, 4 g/ml sinnamik asit kombine dozları ve 2 GyR.ile 1, 2, 4 g/ml sinnamik asit kombine dozlarıyla yapılan ikili karşılaştırmalar sonucunda bir anlamlılık bulunmuştur (p<0,05).Sonuç olarak radyasyonun 1 ve 2 Gy lik dozları COMET ve mikronukleus testlerine göre genotoksik etki göstermiştir. Sinnamik asitte bu radyasyonun indüklediğigenetik hasarı azaltmıştır. COMET ve mikronukleus testlerine dayanarak sinnamikasitin radyoprotektif etkisinin olduğu ortaya konmuştur.
In our study, genotoxic effects of ionizing radiation which is a physical mutagen andradioprotective effects of cinnamic acidare evaluated. For this purpose in vitro COMET and micronucleus assay were carried out in human peripheral blood lymphocytes.Blood sample sare drawn in to tubes with heparin as an anticoagulantandlymphocytes are isolated. Isolated lymphocytes were cultured. After 24 hour followingculture initiation, the lymphocytes were in cubated with 1, 2 and 4g/mluding control lymphocytes were exposed to ionizing radiation. Onceviabilities were ensured, more than 80%, the lymphocytes were subjected to the COMET and micronucleus assay after irradiation. According to COMET assay results,comparisons were made using the values of % Damaged Cells and Genetic Damage Index. In terms of Genetic Damage Index, as a result of the comporisons of 1 Gy.R. with 1, 2 and 4 combined of cinnamic acid and 2 Gy.R. with 1, 2 and 4 combined of cinnamic acid statistically significant difference was found (p<0,001). Using the COMET assay, according to percent age of damaged cells, 1 Gy.R with 1, 2 and 4 combined of cinnamic acid and 2 Gy.R. with 1, 2 and 4 combined of cinnamic acid significant differences were found(p<0,005).According to MN frequency and Nuclear Division Index, statistical comparisons were made using the required test methods. No statistically significant differences were found in terms of NBI (p>0,05). In terms off requency of MN as a result of the comporisons of 1 Gy.R. with 1, 2 and 4 combined of cinnamic acid and 2 Gy.R. with 1, 2 and 4 combined of cinnamic acid significant difference was found (p<0,05).In present study demon strates that 1 and 2 Gy doses of radiation showed genotoxiceffects in human peripheral blood lymphocytes. Ionizing radiation induced geneticdamage is decreased by cinnamic acid. The current study demon stratesth at cinnamicacid is effective in protecting lymphocytes again stradiation-induced toxicity.
In our study, genotoxic effects of ionizing radiation which is a physical mutagen andradioprotective effects of cinnamic acidare evaluated. For this purpose in vitro COMET and micronucleus assay were carried out in human peripheral blood lymphocytes.Blood sample sare drawn in to tubes with heparin as an anticoagulantandlymphocytes are isolated. Isolated lymphocytes were cultured. After 24 hour followingculture initiation, the lymphocytes were in cubated with 1, 2 and 4g/mluding control lymphocytes were exposed to ionizing radiation. Onceviabilities were ensured, more than 80%, the lymphocytes were subjected to the COMET and micronucleus assay after irradiation. According to COMET assay results,comparisons were made using the values of % Damaged Cells and Genetic Damage Index. In terms of Genetic Damage Index, as a result of the comporisons of 1 Gy.R. with 1, 2 and 4 combined of cinnamic acid and 2 Gy.R. with 1, 2 and 4 combined of cinnamic acid statistically significant difference was found (p<0,001). Using the COMET assay, according to percent age of damaged cells, 1 Gy.R with 1, 2 and 4 combined of cinnamic acid and 2 Gy.R. with 1, 2 and 4 combined of cinnamic acid significant differences were found(p<0,005).According to MN frequency and Nuclear Division Index, statistical comparisons were made using the required test methods. No statistically significant differences were found in terms of NBI (p>0,05). In terms off requency of MN as a result of the comporisons of 1 Gy.R. with 1, 2 and 4 combined of cinnamic acid and 2 Gy.R. with 1, 2 and 4 combined of cinnamic acid significant difference was found (p<0,05).In present study demon strates that 1 and 2 Gy doses of radiation showed genotoxiceffects in human peripheral blood lymphocytes. Ionizing radiation induced geneticdamage is decreased by cinnamic acid. The current study demon stratesth at cinnamicacid is effective in protecting lymphocytes again stradiation-induced toxicity.
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Keywords
İyonize radyasyon, Sinnamik asit, Comet testi, Mikronukleus testi, Antioksidan, Genotoksisite, Radyoprotektörler, IonizingRadiation, Cinnamicacid, COMET assay, Micronucleus test, Antioxidant, Genotoxicity, Radioprotection
Citation
Tüzün, E. (2013). Cinnamic (sinnamik) asidin radyasyondan koruyucu etkisinin in vitro kısa süreli genotoksisite test yöntemleri ile araştırılması. Yayınlanmamış yüksek lisans tezi. Uludağ Üniversitesi Fen Bilimleri Enstitüsü.