Türk propolisinin histon deasetilaz (HDAC) enzim aktivitesine etkisinin araştırılması
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Tarih
2021
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Tekirdağ Namık Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bireylerin beslenme ve yaşam tarzı çeşitli epigenetik değişikliklere neden olarak vücudun fizyolojik ve patolojik süreçlerini etkileyebilmektedir. Son yıllarda yapılan çalışmalarda çeşitli besin ve besin bileşenlerinin histon deasetilaz (HDAC) enzim aktivitesini baskıladığına dair güçlü kanıtlar bulunmuştur. Tez çalışmamız kapsamında çeşitli farmakolojik etkilere sahip Türk propolisinin (TP) HEK293T ve HeLa hücre hatlarında HDAC aktivitesi üzerindeki etkisi araştırılmıştır. Çalışmada, Türkiye'de piyasadan toplanan ticari formda, saflaştırılmış 2 farklı propolisin suda çözünen formları kullanılmıştır. HEK293T ve HeLa hücrelerinde HDAC aktivitesinin tespiti için öncelikle TP'nin uygulanması planlanan LD50 konsantrasyonları tripan mavisi boyama yöntemi ile belirlenmiştir. NETN lizis yöntemi ile elde edilen hücresel lizatların protein miktar tayininde Bradford yöntemi kullanılmıştır. In vitro koşullarda Propolis-1'in, LD50 dozu ile hem 24 saatlik muamelesinin hem de doğrudan hücresel lizatlarla muamelesinin, HEK293T ve HeLa hücrelerindeki HDAC aktivitesini azalttığı saptanmıştır. Propolis-2'nin doğrudan muamelesi bilgi verici olmamakla birlikte, 24 saatlik uygulaması ise normal hücrede (HEK293T) HDAC aktivitesini düşürürken kanser hücresinde (HeLa) HDAC aktivitesini artırdığı tespit edilmiştir. Çalışmamızdan elde edilen veriler ışığında TP'nin bir normal hücre hattı olan HEK293T hücrelerinde doğrudan ve dolaylı mekanizmalarla HDACi etkisi olduğu, bir kanser hücre hattı olan HeLa hücrelerinde ise HDAC aktivitesi üzerindeki etkisinin çelişkili olduğu gösterilmiştir.
Nutrition and lifestyle can affect the physiological and pathological processes of the body by causing various epigenetic changes. Strong evidences on various foods and nutritional components suppress histone deacetylase (HDAC) enzyme activity have been found in recent studies. In our thesis study, the effects of Turkish propolis, which has various pharmacological effects, on HDAC activity in HEK293T and HeLa cell lines were investigated. Water-soluble forms of two different purified propolis in commercial form collected from the market in Turkey were used in the study. In order to detect the HDAC activity in HEK293T and HeLa cells, LD50 concentrations, which were planned to be applied for the TP, were determined by trypan blue staining method. Bradford method has been used for protein quantification of cellular lysates prepared by NETN lysis method. It has been determined that both 24-hours treatment and direct treatment of cellular lysates with the Turkish Propolis-1 with the LD50 concentration reduced HDAC activity in HEK293T and HeLa cells in in vitro conditions. Although the results of the cellular lysates treated directly with the Propolis-2 is not informative, the of 24-hours treatment has shown decreasing HDAC activity in normal cell (HEK293T) while increasing HDAC activity in cancer cell (HeLa). Our results show that the TP has HDACi effect in HEK293T cells, a normal cell line, by direct and indirect mechanisms, whereas the findings of HDAC activity in HeLa cells, a cancer cell line, are contradictory.
Nutrition and lifestyle can affect the physiological and pathological processes of the body by causing various epigenetic changes. Strong evidences on various foods and nutritional components suppress histone deacetylase (HDAC) enzyme activity have been found in recent studies. In our thesis study, the effects of Turkish propolis, which has various pharmacological effects, on HDAC activity in HEK293T and HeLa cell lines were investigated. Water-soluble forms of two different purified propolis in commercial form collected from the market in Turkey were used in the study. In order to detect the HDAC activity in HEK293T and HeLa cells, LD50 concentrations, which were planned to be applied for the TP, were determined by trypan blue staining method. Bradford method has been used for protein quantification of cellular lysates prepared by NETN lysis method. It has been determined that both 24-hours treatment and direct treatment of cellular lysates with the Turkish Propolis-1 with the LD50 concentration reduced HDAC activity in HEK293T and HeLa cells in in vitro conditions. Although the results of the cellular lysates treated directly with the Propolis-2 is not informative, the of 24-hours treatment has shown decreasing HDAC activity in normal cell (HEK293T) while increasing HDAC activity in cancer cell (HeLa). Our results show that the TP has HDACi effect in HEK293T cells, a normal cell line, by direct and indirect mechanisms, whereas the findings of HDAC activity in HeLa cells, a cancer cell line, are contradictory.
Açıklama
Anahtar Kelimeler
Beslenme ve Diyetetik, Nutrition and Dietetics, Türk Propolisi, HDACi, Epigenetik, Fonksiyonel Besin., Turkish Propolis, HDACi, Epigenetic, Functional Food.
