İn Vitro Hepatik Oksidatif Hasarda Karvakrolün Etkinliğinin Araştırılması

Öz

Nonalkolik karaciğer yağlanması (NAFLD) karaciğerde, özellikle trigliserit olmak üzere yağ birikimi ile karakterize, yaygın görülen bir hastalıktır. Hastalığın tedavisinde araştırmalar, doğal kaynaklardan elde edilen antioksidan moleküllere yoğunlaşmıştır. Bu bağlamda çalışmamızda, kekiğin etken maddesi karvakrolün in vitro hepatoprotektif etkinliği araştırılmıştır. Araştırmamızda materyal olarak insan hepatosit hücre hattı (HepG2, ATCCHB-8065) kullanılmıştır. Deneme grupları kontrol (K), palmitat eklenen grup (P), karvakrol eklenen grup (C) ve palmitat ile birlikte karvakrol eklenen grup (+C) olarak tasarlanmıştır. Farklı konsantrasyonlarda palmitat (150, 300 ve 450 µM) ve karvakrol (1, 5, 10 ve 50 µM) 24 saat sürelerle hücrelere uygulanarak etkin konsantrasyon tespitleri MTT viyabilite testi ile ortaya konulmuştur. Hücrelerden elde edilen lizatlardan hücre içi glutatyon ve nitrit düzeyi spektrofotometrik olarak analiz edilmiştir. Yine bu örneklerdeki aldoz redüktaz düzeyleri (AR) ELISA metoduyla araştırılmıştır. Çalışmadan elde edilen viyabilite verilerine göre, palmitatın 300 µM konsantrasyonunun kontrole göre % 45 oranında hücre kayıplarına neden olduğu, karvakrolün 5 µM konsantrasyonunun hücrelere uygulanımıyla bu kayıpların % 41 oranında önlendiği tespit edilmiştir. Çalışmada palmitat ve karvakrolün GSH düzeyinde önemli bir değişiklik oluşturmadığı ancak NO düzeylerini anlamlı düzeyde azalttığı tespit edilmiştir. Yine hücre içi AR protein düzeyini palmitat arttırırken, karvakrolün bu artışı % 1.7 oranında önlediği belirlenmiştir. Tüm bu veriler doğrultusunda karvakrolün nonalkolik steatozis gibi yağlanmaya bağlı karaciğer hastalıklarında potansiyel bir etken madde olarak düşünülebileceği kanısına varılmıştır.

Anahtar Kelimeler

• Nonalkolik karaciğer yağlanması
• palmitat
• karvakrol
• oksidatif stres
• aldoz redüktaz

Investigation of the Efficacy of Carvacrol in In Vitro Hepatic Oxidative Damage

Öz

Non-alcoholic fatty liver disease (NAFLD); is a common disease characterized by accumulation of fat in the liver, especially triglycerides. Research in the treatment of the disease has focused on antioxidant molecules derived from natural sources. In this context, hepatoprotective effect of carvacrol has been tried to be demonstrated by in vitro experiments. Human hepatocyte cell line (HepG2, ATCCHB-8065) was used in the study. Experimental groups were designed as control (K), palmitate added group (P), carvacrol added group (C) and palmitate and carvacrol added group (+ C). Different concentrations of palmitate (150, 300 and 450 µM) and carvacrol (1, 5, 10 and 50 µM) were applied to the cells for 24 hours and effective concentration determinations were determined by MTT viability test. Intracellular glutathione and nitrite levels were analyzed spectrophotometrically from lysates obtained from cells. Besides, aldose reductase levels (AR) in these samples were investigated by ELISA method. According to the viability data obtained from the study, it was found that 300 µM concentration of palmitate caused 45% cell losses compared to the control, and 5 µM concentration of carvacrol prevented 41% of these cell losses. In this study, palmitate and carvacrol did not produce a significant change in GSH levels but it significantly reduced NO levels. In addition, palmitate increased intracellular AR protein level, while carvacrol inhibited this increase by 1.7%. According to all these data, it was concluded that carvacrol can be considered as a potential active agent in fatty liver diseases such as nonalcoholic fatty liver disease.

Anahtar Kelimeler

• Nonalcoholic fatty liver disease
• palmitate
• carvacrol
• oxidative stress
• aldose reductase

Kaynakça

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