Hepatoprotective effect of Eugenol on Acetaminophen-Induced Hepatotoxicity in HepG2 cells

Imbalance in liver metabolism lead to oxidative stress mainly caused by free radicals or termed as reactive oxidative oxygen (ROS). Prolonged ROS exposure without proper treatment induce severe liver damage and serious hepatic diseases including cirrhosis. Eugenol (4-allyl 2-methoxyphenol) is phenolic derivative compound that showed antioxidant, anti-inflammatory, analgesic, antibacterial, antifungal, and antitumor activities. This study aims to evaluate the hepatoprotective effect of eugenol through biochemical markers analysis. Cytotoxic assay was performed in various concentrations of eugenol (3,125; 6,25; 12,5; 25; 50; 100 μg/mL) using (3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxyphenkyl)-2-(4-sulfophenyl)-2H-tetrazolium) to determine the safe concentrations for next assays. Aspartate aminotransferase (AST), alanin aminotransferase (ALT), and lactate dehydrogenase (LDH) assay were performed using colorimetric method to evaluate the levels and activity of liver-related enzymes which are elevated in damaged liver as they were used as hepatotoxicity markers. The viability of HepG2 cells increased in eugenol concentration 3.125 μg/mL and then decreased along with the rise of eugenol concentrations. From this cytotoxic assay, two concentrations of eugenol were choosen (6.25 and 25 ug/ml) to be evaluated in the next assays. The level of LDH, ALT, and AST decreased after eugenol treatment compared to negative control. The most effective concentration of eugenol to seemed different in certain hepatotoxicity markers. This study suggests that eugenol was safe to use for cells culture environment in large ranges of concentrations and shows hepatoprotective effect in APAP-induced hepatotoxicity model by the decrease of LDH level and AST and ALT activities.