Investigation of possible protective effects of propolis extract against bile acid induced hepatotoxicity in C3A cell line

Cholestasis is a pathophysiological condition that characterized by the accumulation in the liver of bile salts, biliary pressure increase, bile duct occlusion. Propolis, a bee products, is rich about polyphenolic compounds and flavonoids. İts antitumor, anti-inflammatory, anti-oxidative, antimutagenic properties were researched. We aimed to possible protective effect of propolis extract against bile acid induced hepatotoxicity on viability of hepatocyte. We used C3A human hepatoma cell line in this study. Cells were grown in DMEM/F12 medium containing %10 FCS and %1 antibiotic under condition with humidified atmosphere (37°C, %5 CO2). Cells were harvested using trypsin/EDTA and were counted with a hemocytometer. Cells were seeded at concentration of 3x104 hücre/100µl into 96 well plate. We allowed for all cells attach to plate during a day. GCDCA (concentration of 0.5, 0.75, 1, 1.5, 2 mM) was added in medium of cells (n=3). After 24h administration of the bile acid, we determinated toxic dose of GCDCA is 1 mM by MTT assay. Ethanol and aqueous extracts of propolis (concentration of 50, 10, 5, 1 µg/ml) were administered before and after 3h treatment of determinated effective dose of bile acid. Possible protective effects of propolis were measured by using MTT assay 24h later. Ethanol content of propolis solution applied on cells was kept below 0.05%. The cell morphology was evaluated by acridine orange staining under the fluorescent microscope. Comparing alone treatment group of ethanol and aqueous propolis extracts with the control group did not show significant effects (p>0.05). It shown that all dose of aqueous extract of propolis treatment before toxicity induced by 1 mM GCDCA and 10 to 50 µg/ml aqueous extract of propolis after toxicity induced by 1 mM GCDCA significantly increased cell viability(p<0.05). Before toxicity induced by 1.5 mM GCDCA, 10 to 50 µg/ml aqueous extract of propolis shown positively effect on cell viability(p<0.05). Before and after toxicity induced by 1 mM GCDCA, 1 µg/ml ethanol extract of propolis shown positively effect(p<0.05). Before and after toxicity induced by 1.5 mM GCDCA, ethanol extract of propolis did not show positively effect(p>0.05). Because of to be scarcely therapy for protection of the liver during the waiting period before operation of cholestasis and to be seen in-vitro protective effect of propolis by this study, in-vivo study about this event is reqiured.