Biyoaktif besin bileşeni piperlongumin'in histon deasetilaz enzim inhibitörü etkisinin araştırılması
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Dosyalar
Tarih
2021
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Yayıncı
Tekirdağ Namık Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Beslenme ve Diyetetik alanında önemli bir yeri olan besin-gen etkileşimlerinin araştırılmasıyla besinlerin organizma üzerindeki etkilerinin daha iyi anlaşılması hedeflenmektedir. Son dönemde yapılan çalışmalarla çeşitli besinlerdeki çok sayıda biyoaktif bileşen Histon Deasetilaz Enzimi İnhibitörü (HDACi) olarak tanımlanmıştır. Bu tez çalışması kapsamında, biyoaktif bir besin bileşeni olarak bilinen Piperlongumin'in (PL) HeLa ve HEK293T hücre soylarındaki HDACi etkisi deneysel olarak araştırılmıştır. PL'in HeLa ve HEK293T hücre soylarındaki HDACi etkisini tespit etmek için, öncelikle uygulanması planlanan LD50 dozları MTT ve AO/PI boyama yöntemleriyle tespit edilmiştir. NETN lizis yöntemi ile hazırlanan hücresel lizatlarda protein miktar tayini Bradford yöntemiyle gerçekleştirilmiştir. Yürütülen deneysel çalışmalar ile; HDAC enzim aktivitesinin, 24 ve 48 saat PL uygulanması sonrasında HeLa hücrelerinde azaldığı, HEK293T hücrelerinde ise arttığı tespit edilmiştir. Normal koşullarda çoğaltılan HeLa ve HEK293T hücrelerinden hazırlanan lizatlar HDAC aktivite tayininden hemen önce PL ile doğrudan muamele edildiğinde ise, HDAC enzim aktivitesinin HeLa hücrelerinde istatistiksel açıdan anlamlı olarak arttığı, HEK293T hücrelerinde ise azaldığı belirlenmiştir. Sonuçlarımız PL'in HEK293T normal ve HeLa kanser hücre soylarında HDAC enzim aktivitesi üzerine farklı yönde etkisinin olduğunu göstermiştir. Ayrıca sonuçlarımız HEK293T ve HeLa hücre soyları, hücre kültürü ortamında PL ile 24 ila 48 saat sürelerle muamele edildiğinde veya PL hücresel lizatlarla doğrudan muamele edildiğinde yine zıt yönde etkiler oluşabileceğini ortaya koymuştur. Elde edilen sonuçlar PL ile HDAC enzimleri arasında doğrudan bir etkileşim olabileceği gibi PL' in HDAC enzim aktivitesi üzerinde dolaylı mekanizmalarla da etkili olabileceğini ortaya koymaktadır.
It is aimed to better understand the effects of nutrients on the organism by investigating the nutrient-gene interactions that have an important place in the field of Nutrition and Dietetics. In recent studies, many bioactive components in various foods have been identified as Histone Deacetylase Enzyme Inhibitor (HDACi). In this thesis, HDACi effect of Piperlongumine (PL), known as a bioactive nutrient component, on HeLa and HEK293T cell lines was experimentally investigated. In order to determine the effect of PL on HDAC activity in HeLa and HEK293T cell lines, the LD50 doses planned to be applied were firstly determined by MTT and AO/PI staining methods. Protein quantification in cellular lysates prepared by the NETN lysis method was carried out by the Bradford assay. With the experimental studies carried out in this study, it was determined that HDAC enzyme activity decreased in HeLa cell lines and increased in HEK293T cell lines after 24 and 48 hours of PL applied. When lysates prepared from HeLa and HEK293T cells grown under normal conditions were treated directly with PL just before HDAC activity assay, it was determined that HDAC enzyme activity increased statistically significantly in HeLa cells and decreased in HEK293T cells. Our results showed that PL has a different effect on HDAC enzyme activity in HEK293T normal and HeLa cancer cell lines. In addition, our results demonstrated that the opposite effects can also occur when HEK293T and HeLa cell lines are applied with PL for 24 to 48 hours in cell culture medium or when cellular lysates applied directly with the PL. The results obtained show that there may be a direct interaction between PL and HDAC enzymes, also PL can be effective with indirectly mechanisms on HDAC enzyme activity.
It is aimed to better understand the effects of nutrients on the organism by investigating the nutrient-gene interactions that have an important place in the field of Nutrition and Dietetics. In recent studies, many bioactive components in various foods have been identified as Histone Deacetylase Enzyme Inhibitor (HDACi). In this thesis, HDACi effect of Piperlongumine (PL), known as a bioactive nutrient component, on HeLa and HEK293T cell lines was experimentally investigated. In order to determine the effect of PL on HDAC activity in HeLa and HEK293T cell lines, the LD50 doses planned to be applied were firstly determined by MTT and AO/PI staining methods. Protein quantification in cellular lysates prepared by the NETN lysis method was carried out by the Bradford assay. With the experimental studies carried out in this study, it was determined that HDAC enzyme activity decreased in HeLa cell lines and increased in HEK293T cell lines after 24 and 48 hours of PL applied. When lysates prepared from HeLa and HEK293T cells grown under normal conditions were treated directly with PL just before HDAC activity assay, it was determined that HDAC enzyme activity increased statistically significantly in HeLa cells and decreased in HEK293T cells. Our results showed that PL has a different effect on HDAC enzyme activity in HEK293T normal and HeLa cancer cell lines. In addition, our results demonstrated that the opposite effects can also occur when HEK293T and HeLa cell lines are applied with PL for 24 to 48 hours in cell culture medium or when cellular lysates applied directly with the PL. The results obtained show that there may be a direct interaction between PL and HDAC enzymes, also PL can be effective with indirectly mechanisms on HDAC enzyme activity.
Açıklama
Anahtar Kelimeler
Beslenme ve Diyetetik, Nutrition and Dietetics
