Mirisetin'in insan lenfosit DNA'sı üzerindeki radyoprotektif etkisinin incelenmesi
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Dosyalar
Tarih
2013
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Trakya Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, bir flavonoid olan Mirisetin'in insan lenfosit DNA'sı için radyoprotektif olup olmadığı araştırılmıştır. Bu amaçla, gönüllülerden alınan kan örnekleri ile Mikronukleus, Komet ve ELİSA deneyleri yapılmıştır. Her deney serisi için kontrol, DMSO, 100 µM ve 200 µM Mirisetin, 0.5 Gy ve 2 Gy radyasyon dozları ile Mirisetin ve radyasyonun birlikte uygulamalarının sonuçları analiz edilmiştir. Mikronukleus deneyleri için kan örnekleri Mirisetin ve radyasyon uygulamalarını takiben 68 saat kültüre alınmış ve sitokinez aşamasında Sitokalazin-B ile durdurulan hücreler ışık mikroskobu ile değerlendirilmiştir. Deney sonuçlarına göre, 100 µM ve 200 µM Mirisetin uygulamasının 0.5 Gy ve 2 Gy dozlarda radyasyonun hasar verici etkisini azalttığı gözlenmiştir. Komet deneylerinde 2 farklı protokol uygulanmıştır: Alkali ve FPG Komet yöntemleri. pH>13 Alkali Komet DNA?daki tek ve çift zincir kırıklarını tespit etmek için, FPG komet ise radyasyon nedeni ile meydana gelen oksidatif DNA hasarlarını belirlemek amacıyla kullanılmıştır. Komet deney sonuçlarına göre, 100 µM ve 200 µM dozlarda Mirisetin'in kendisinin de DNA'da ileri düzeyde kırıklara neden olduğu tespit edilmiştir. Bu nedenle, bu çalışmada uygulanan deney koşulları ile Mirisetin'in radyoprotektif etkisinin değerlendirilmesinde sağlıklı bir sonuç elde edilemeyeceği kanısına varılmıştır.
Abstract
In this study, it is investigated that Myricetin which is a flavonoid is whether radioprotective for human lymphocyte DNA or not. For this purpose, Micronucleus, Comet and ELISA assays were carried out with blood samples were taken from volunteers. For each experiment series, results were analyzed as control, DMSO, 100 µM and 200 µM Myricetin, myricetin with doses of 0.5 Gy and 2 Gy irradiation together. Blood samples were cultured for 68 hours followed by myricetin and radiation applications and cultures were blocked by Cytochalasin-B at cytokinesis stage and cells were evaluated by light microscopy for Micronucleus assays. According to the test results, it was observed that the application of 100 µM and 200 µM Myricetin at 0.5 Gy and 2 Gy doses reduce damaging effect of radiation . Two different protocols were applied in Comet Assays: Alkaline and FPG Comet methods. pH>13 Alkaline Comet was used to detect single and double strand breaks of DNAs, as for FPG Comet, it was used to determine oxidative DNA damage induced by radiation. According to Comet Assay results, it was determined that myricetin also itself causes breaks highly on DNA at 100 µM and 200 µM doses. Therefore, it was concluded that no accurate results is able to obtain to evaluate the radioprotective effect of Myricetin in the test conditions applied in these assays.
Abstract
In this study, it is investigated that Myricetin which is a flavonoid is whether radioprotective for human lymphocyte DNA or not. For this purpose, Micronucleus, Comet and ELISA assays were carried out with blood samples were taken from volunteers. For each experiment series, results were analyzed as control, DMSO, 100 µM and 200 µM Myricetin, myricetin with doses of 0.5 Gy and 2 Gy irradiation together. Blood samples were cultured for 68 hours followed by myricetin and radiation applications and cultures were blocked by Cytochalasin-B at cytokinesis stage and cells were evaluated by light microscopy for Micronucleus assays. According to the test results, it was observed that the application of 100 µM and 200 µM Myricetin at 0.5 Gy and 2 Gy doses reduce damaging effect of radiation . Two different protocols were applied in Comet Assays: Alkaline and FPG Comet methods. pH>13 Alkaline Comet was used to detect single and double strand breaks of DNAs, as for FPG Comet, it was used to determine oxidative DNA damage induced by radiation. According to Comet Assay results, it was determined that myricetin also itself causes breaks highly on DNA at 100 µM and 200 µM doses. Therefore, it was concluded that no accurate results is able to obtain to evaluate the radioprotective effect of Myricetin in the test conditions applied in these assays.
Açıklama
Doktora Tezi
Anahtar Kelimeler
Mirisetin, Radyoprotektif Etki, Komet Deneyi, Mikronukleus, Myricetin, Radioprotective Effect, Comet Assay, Micronucleus
