A Comparative Study of the Hepatoprotective Effect of Centella asiatica Extract (CA-HE50) on Lipopolysaccharide/d-galactosamine-Induced Acute Liver Injury in C57BL/6 Mice

Abstract

Acute liver failure (ALF) refers to the sudden loss of liver function and is accompanied by several complications. In a previous study, we revealed the protective effect of Centella asiatica 50% ethanol extract (CA-HE50) on acetaminophen-induced liver injury. In the present study, we investigate the hepatoprotective effect of CA-HE50 in a lipopolysaccharide/galactosamine (LPS-D-Gal)-induced ALF animal model and compare it to existing therapeutic silymarin, Lentinus edodes mycelia (LEM) extracts, ursodeoxycholic acid (UDCA) and dimethyl diphenyl bicarboxylate (DDB). Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were decreased in the CA-HE50, silymarin, LEM, UDCA and DDB groups compared to the vehicle control group. In particular, AST and ALT levels of the 200 mg/kg CA-HE50 group were significantly decreased compared to positive control groups. Lactate dehydrogenase (LDH) levels were significantly decreased in the CA-HE50, silymarin, LEM, UDCA and DDB groups compared to the vehicle control group and LDH levels of the 200 mg/kg CA-HE50 group were similar to those of the positive control groups. Superoxide dismutase (SOD) activity was significantly increased in the 100 mg/kg CA-HE50, LEM and UDCA groups compared to the vehicle control group and, in particular, the 100 mg/kg CA-HE50 group increased significantly compared to positive control groups. In addition, the histopathological lesion score was significantly decreased in the CA-HE50 and positive control groups compared with the vehicle control group and the histopathological lesion score of the 200 mg/kg CA-HE50 group was similar to that of the positive control groups. These results show that CA-HE50 has antioxidant and hepatoprotective effects at a level similar to that of silymarin, LEM, UDCA and DDB, which are known to have hepatoprotective effects; further, CA-HE50 has potential as a prophylactic and therapeutic agent in ALF.

Keywords: Centella asiatica; acute liver failure; antioxidant; hepatoprotective effect.

Figure 1

Effects of CA-HE50 on body weight and food consumption changes in LPS/D-Gal-induced ALF animal model. (A) The body weights of mice in eight different experimental groups were measured on days 0, 8, 14 and 15. (B) Food consumption of each experimental group was measured once per week during the entire experiment. NC, normal control; VC, vehicle control; CA100, 100 mg/kg CA-HE50; CA200, 200 mg/kg CA-HE50; Silymarin, 100 mg/kg silymarin; LEM, 200 mg/kg LEM; UDCA, 25 mg/kg UDCA; DDB, 200 mg/kg DDB. Data are represented as mean ± SD.

Figure 2

Effects of CA-HE50 on relative organ weight changes in LPS/D-Gal-induced ALF animal model. The weights of major organs collected from mice in each group were measured. (A) Heart; (B) liver; (C) lung; (D) spleen; (E) kidney. NC, normal control; VC, vehicle control; CA100, 100 mg/kg CA-HE50; CA200, 200 mg/kg CA-HE50; Silymarin, 100 mg/kg silymarin; LEM, 200 mg/kg LEM; UDCA, 25 mg/kg UDCA; DDB, 200 mg/kg DDB. Data are represented as mean ± SD (* p < 0.05 or ** p < 0.01 vs. vehicle control).

Figure 3

Effects of CA-HE50 on liver damage and blood lipids in LPS/D-Gal-induced ALF animal model. The serum levels of (A) AST, (B) ALT, (C) GGT, (D) LDH, (E) TG, (F) TC, (G) HDL and (H) LDL were investigated. NC, normal control; VC, vehicle control; CA100, 100 mg/kg CA-HE50; CA200, 200 mg/kg CA-HE50; Silymarin, 100 mg/kg silymarin; LEM, 200 mg/kg LEM; UDCA, 25 mg/kg UDCA; DDB, 200 mg/kg DDB. Data are represented as mean ± SD (^## p < 0.01 vs. normal control; * p < 0.05 or ** p < 0.01 vs. vehicle control).

Figure 4

Effects of CA-HE50 on CAT and SOD activities in LPS/D-Gal-induced ALF animal model. The activity changes of representative antioxidant enzymes (A) CAT and (B) SOD in liver tissue were investigated. NC, normal control; VC, vehicle control; CA100, 100 mg/kg CA-HE50; CA200, 200 mg/kg CA-HE50; Silymarin, 100 mg/kg silymarin; LEM, 200 mg/kg LEM; UDCA, 25 mg/kg UDCA; DDB, 200 mg/kg DDB. Data are represented as mean ± SD (^## p < 0.01 vs. normal control; * p < 0.05 vs. vehicle control).

Figure 5

Effect of CA-HE50 on histopathological changes in LPS/D-Gal-induced ALF animal model. Histopathological changes in the extracted liver tissue were confirmed by H&E and TUNEL analysis. Histopathological changes in liver tissue were quantified by lesion scores according to the severity of the lesion. (A) Representative images of each group in the histopathological lesion. Histopathological lesion scores are presented as a bar diagram. (B) Representative images of each group in apoptotic dead cells by TUNEL analysis. The numbers of apoptotic dead cells are presented as a bar diagram. NC, normal control; VC, vehicle control; CA100, 100 mg/kg CA-HE50; CA200, 200 mg/kg CA-HE50; Silymarin, 100 mg/kg silymarin; LEM, 200 mg/kg LEM; UDCA, 25 mg/kg UDCA; DDB, 200 mg/kg DDB. Data are represented as mean ± SD (^## p < 0.01 or ^### p < 0.001 vs. normal control; * p < 0.05, ** p < 0.01 or *** p < 0.001 vs. vehicle control).