Anti-oxidative and hepatoprotective effects of lithospermic acid against carbon tetrachloride-induced liver oxidative damage in vitro and in vivo

Accumulation of an excess amount of reactive oxygen species (ROS) can cause hepatotoxicity that may result in liver damage. Therefore, development of anti-oxidative agents is needed for reducing liver toxicity. This study investigated the anti-oxidative and hepatoprotective activity of lithospermic acid, a plant-derived polycyclic phenolic carboxylic acid isolated from Salvia miltiorrhiza, on carbon tetrachloride (CCl4)-induced acute liver damage in vitro and in vivo. The results of the DPPH assay indicated that lithospermic acid was a good anti-oxidant. The CCl4-exposed Huh7 cell line exhibited decreased cell viability, increased necrosis and elevated ROS and caspase-3/7 activity. Lithospermic acid significantly attenuated the CCl4-induced oxidative damage in a concentration-dependent manner. The result of an in vivo study with BALB/c mice corresponded with the anti-oxidative activity noted in the in vitro study. Exposure of mice to CCl4 resulted in a greater than 2-fold elevation in serum aspartate transaminase (AST) and alanine transaminase (ALT). levels In addition, CCl4-intoxication led to an over 20% decrease in the level of intracellular hepatic enzymes including superoxide dismutase (SOD) and catalase (CAT) as well as increased lipid peroxidation. Upon histological examination of the CCl4-exposed mice, the mouse livers showed severe hepatic damage with a huge section of necrosis and structural destruction. Pretreatment of mice with lithospermic acid for six days significantly reduced CCl4-induced hepatic oxidative damage, serum AST and ALT. The pretreatment also increased SOD and CAT. The findings suggest that the health status of the liver was improved comparable to the control group after a high-dose treatment with lithospermic acid (100 mg/kg weight). The potential applicability of lithospermic acid as a hepatoprotective agent was demonstrated.