İn Vitro Lipopolisakkarit Uyarımlı Karaciğer Yangı Modelinde Alfa-Terpineol’ün Etkinliğinin Araştırılması

Öz

Terpinoller monoterpen bileşikler olup, birçok çalışmada antioksidan özelliklerinden dolayı antikanser, antikonvülsant ve antiülser gibi biyolojik etkileri ortaya konulmuştur. Araştırmanın amacını, LPS’in karaciğer hücrelerine uygulanarak oluşturulan yangı modelinde, α-terpineol’ün antiinflamatuar ve antiapoptotik biyofonksiyonlarının araştırılması oluşturmuştur. Denemede insan orijinli HepG2 (ATCC® HB-8065) hücreleri materyal olarak seçilmiştir. LPS ve α-terpineol, hücrelere farklı konsantrasyonlarda 24 saat uygulanmıştır ve etkin konsantrasyonları, hücre canlılık testleriyle (MTT) gerçekleştirilmiştir. Sonrasında, TNF-α, IL-1β, IL-10, Kaspaz 3, Bax ve Bcl-2 gen ekspresyon düzeyleri tam zamanlı kantatif polimeraz zincir reaksiyonuyla (qRT-PCR) araştırılmıştır. LPS’in 50 ng/ml konsantrasyonu %11,5 oranında hücre kayıplarına neden olduğu belirlenmiş ve modelleme için bu konsantrasyon seçilmiştir. Ancak, 10 μM konsantrasyonda α-terpineolün hücre kayıplarını % 2.12 oranında önlediği bulunmuştur. Çalışmada, LPS’in TNF-α ve IL-1β gen ekspresyonlarını arttırdığı, α-terpineol uygulamasının ise bu durumu tersine çevirdiği tespit edilmiştir. Yine, IL-10 gen ekspresyonu yüksek LPS konsantrasyonu ile baskılanırken, α-terpineol tarafından anlamlı düzeyde uyarıldığı da ortaya konulmuştur. Bununla birlikte, LPS’in kaspaz 3 ve Bax gen ekspresyonlarını arttırdığı, ancak α-terpineol’ün bu stimülasyonu baskıladığı tespit edilmiştir. Bcl-2 gen ekspresyonları ise LPS tarafından baskılanırken, α-terpineol tarafından uyarıldığı bulunmuştur. Özetle, α-terpineol’ün kısa süreli ve düşük olan proliferatif konsantrasyonunun özellikle patojen kaynaklı karaciğer rahatsızlıklarında alternatif bir tedavi ajanı olabileceği görülmüştür.

Anahtar Kelimeler

• α- terpineol
• apoptoz
• inflamasyon
• lipopolisakkarit

Investigation of The Efficiency Of Alpha-Terpineol in the In Vitro Lipopolisacharide Stimulated Liver Inflammation Model

Öz

Terpinols are monoterpene compounds, and biological effects like anticancer, anticonvulsant and antiulcer have been demonstrated in many studies due to their antioxidant properties. The aim of the research was to investigate the anti-inflammatory and antiapoptotic biofunctions of α-terpineol in LPS induced inflammation model of liver cells. HepG2 (ATCC® HB-8065) cells of human origin were selected as material. LPS and α-terpineol were applied to cells at different concentrations for 24 hours, and their effective concentrations were determined by cell viability tests (MTT). Afterwards, gene expression levels of TNF-α, IL-1β, IL-10, Caspase 3, Bax and Bcl-2 were investigated by qRT-PCR method. LPS at 50 ng/ml, caused 11.5% cell loss and was chosen for modeling. However, α-terpineol at 10 μM concentration prevented 2.12% of cell loss. In the study, LPS increased TNF-α and IL-1β gene expressions, and α-terpineol application reversed this situation. Again, while IL-10 was suppressed by high LPS concentration, it was significantly stimulated by α-terpineol. Besides, LPS increased caspase 3 and Bax gene expressions, but α-terpineol suppressed this stimulation. Bcl-2 gene expressions were suppressed by LPS and stimulated by α-terpineol. In summary, short-term and low proliferative concentration of α-terpineol can be an alternative treatment agent, especially in pathogen-induced liver disorders.

Anahtar Kelimeler

• α-terpineol
• apoptosis
• inflammation
• lipopolysaccharide

Kaynakça

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