Wu-Chia-Pi solution attenuates carbon tetrachloride-induced hepatic injury through the antioxidative abilities of its components acteoside and quercetin

Abstract

Wu-Chia-Pi medicated wine, composed nine Chinese medicines soaked in 35% alcohol, is widely used in Asia for its health-promoting functions. However, long-term consumption of alcohol could result in liver dysfunction. In this study, Wu-Chia-Pi solution (WCPS) and extract (WCPE) were prepared by modification of the principals given by the Committee on Chinese Medicine and Pharmacy in Taiwan. The aim of this study was to explore the protective effect of WCPS against carbon tetrachloride (CCl4)-induced liver injury and to clarify its active component(s). Antioxidative effects of the test samples were evaluated via MDA inhibition, catalase activity and DPPH-scavenging assays. HPLC was used to analysis the active components. Results showed that WCPS (1 and 5 mL/kg) significantly prevented CCl4-induced liver injury without chronic liver toxicity. Referring to the antioxidative activities, WCPE displayed significant MDA inhibitory and DPPH-scavenging activities with IC50 values of 0.91 ± 0.03 and 0.60 ± 0.04 mg/mL, respectively. Catalase activity was also enhanced by treatment of WCPE, acteoside and quercetin. Therefore, we suggest that acteoside and quercetin are the major contributors to the antioxidative and hepatoprotective activities of WCPS, and a possible mechanism could be mediated through reduction of oxidative stress.

Figure 1

Histological analysis of the chronic liver toxicity of WCPS. Liver tissues were stained with hematoxylin and eosin at 40× and 100× magnifications. The dose of WCPS was 5.0 mL/kg/day.

Figure 2

WCPS reduced CCl₄-induced hepatic injury as evidenced by the lower serum GOT (A) and GPT (B) levels. ^# p < 0.05, compard to the sham group. * p < 0.05, compared to the control group, n = 5. Statistical significant differences were determined by One-Way ANOVA followed by a Fisher LSD post hoc test.

Figure 3

MDA inhibitory effects (A) and catalsed activity (B) of WCPE and marker substances. A, WCPE; Control, untreated group. Concentrations of WCPE and marker substances in catalase assay were 10 mg/mL and 5 μM, respectively. * p < 0.05, ** p < 0.005, compared to the control group. Statistical significant differences were determined by One-Way ANOVA followed by a Fisher LSD post hoc test.

Figure 4

The HPLC fingerprint of WCPE (A) and the chemical structures of acteoside and quercetin (B). Retention times of acteoside and quercetin were 8.9 and 22.5 min, respectively. The horizontal axis represented time in minutes, and vertical axis represented mAU, the intensity of the UV absorbance values.