Banu KUBULAY, Altuğ KÜÇÜKGÜL, Azime KÜÇÜKGÜL

Deneysel Yangı Oluşturulmuş Gökkuşağı Alabalıklarında (Oncorhynchus mykiss Walbaum, 1792) Timolün Apoptotik Antiinflamatuvar Etkinliği

Enfekte balık dokusunda apoptotoz ve inflamasyon içerikli gen ekspresyonları üzerine timolün etkinliğini araştıran bu çalışmada 4 farklı grup oluşturuldu. Kontrol (C) ve yalnızca timol destekli yemleme grubu (THY-100 µg ml-1 yem) hariç, tüm balıklar E.coli LPS’i (LPS-25 µg ml-1) ile enfekte edildi. Son grup enfekte balıkların timol destekli yem ile beslenmesi (+THY) ile oluşturuldu. 3 günlük deneme süresi sonunda karaciğer ve böbrekten doku örnekleri alındı. Apoptotik sitokinlerden kaspaz 3 (Cas 3), kaspaz 8 (Cas 8); pro-inflamator sitokinlerden ise interferon gama (IFN-γ) ve interlökin 1 beta (IL-1β) gen ekspresyon seviyeleri Real-Time PCR analizleriyle araştırıldı. LPS uygulanması ile IL-1β ve IFNγ gen düzeylerinde azalmaların olduğu fakat THY ile tekrar kontrol seviyelerine ulaştığı belirlendi. +THY grubunun LPS ile karşılaştırılmalı sonuçları gösterdi ki tüm gen seviyeleri yukarı regülasyonlar ile uyarıldı. LPS enfeksiyonu Cas 3 ve 8 ekspresyon seviyelerinde karaciğerde artışlara, böbrekte ise azalmalara neden oldu. Ancak, sadece timol içeren yemle beslenen (THY) balıklarda herhangi bir değişim gözlenmedi. +THY, kaspaz içerikli tüm genlerde yaklaşık 3 misli yukarı regülasyonlar tespit edildi. Çalışmada, timolün pro-inflamator ve apoptosis içerikli gen seviyelerinde değişimler göstererek etkili olduğu belirlendi. Ancak yinede enfekte balıklar üzerinde bitkisel destekli yemlemenin immün modülasyon üzerindeki etkilerini daha iyi anlamak için daha kapsamlı moleküler çalışmalara gereksinim duyulmaktadır.

Anahtar Kelimeler:

Gökkuşağı Alabalığı, Apoptotik sitokinler, Lipopolisakkarit, Timol, Pro inflamator, sitokinler

Activity of Apoptotic and Antiinflammation on Rainbow Trout (Oncorhynchus mykiss Walbaum, 1792) Inflammation-Induced Experimentally

In this study investigated thymol effects on gene expressions relative to pro-inflammatory and apoptosis in tissue fish infected four different trials were formed. All fish were infected with E. coli LPS (25 µg ml-1) expect control and only thymol (THY) supplementation (100 µg ml-1 diet) group fish. The last group was the infected fish fed with thymol supplementation diet (+THY). Kidney and liver tissues were removed 3 day after the experiment period. The gene expression levels of interleukin-1β (IL-1β) and interferon gamma (INF-γ) from pro-inflammatory cytokines; caspase 3 (Cas 3) and caspase 8 (Cas 8) from apoptotic cytokines were investigated by Real-Time PCR analyses. Decreases recorded on gene levels of IL-1β and IFNγ by LPS treatment, however it was observed that returned to the control level. The comparative results of LPS and +THY treatment showed that all gene levels significantly stimulated by up regulation. The expression levels of Cas 3 and 8 following LPS caused the increases in liver, no in kidney. However, any change was not observed in only diet application (THY). +THY application showed about 3-fold up-regulations on all gene expressions including apoptosis. It was determined to be effective by showing changes on gene levels including pro-inflammatory and apoptosis. However, further studies are required to better understand immune modulatory effect of diet supplementation feeding on the virulence mechanism of fish-infected.

Keywords:

Rainbow trout, Apoptotic cytokines, LPS, Thymol, Proinflammatory, Cytokines,

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