The effects of high-dose qinggan huoxue recipe on acute liver failure induced by d-galactosamine in rats

Abstract

Qinggan Huoxue Recipe is a traditional Chinese medicine, which has been usually used to improve liver function in hepatitis. In order to investigate the effects of high-dose Qinggan Huoxue Recipe on acute liver failure and explore the potential mechanism, we had built acute liver failure models in rats by intraperitoneal injection of D-galactosamine (D-GalN). High-dose Qinggan Huoxue Recipe was delivered by gavage. After treatment, the blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), albumin (ALB), cholinesterase (CHE), and prothrombin time (PT) were determined. The pathological score of liver tissue was recorded. Proliferating cell nuclear antigen (PCNA) immunohistochemistry staining and fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR) of high mobility group box 1 (HMGB1), toll-like receptor 4 (TLR4), nuclear factor-kappa B (NF- κ B), and Caspase-3 were performed. The survival curve was also depicted. Our results demonstrated that high-dose Qinggan Huoxue Recipe could significantly improve liver function and increase survival rates in rats with acute liver failure. These effects were supposed to be mediated by suppressing inflammatory reaction and apoptosis.

Figure 1

The serum ALT, AST, TBIL, ALB, and CHE levels and PT after treatment. (a) ALT; (b) AST; (c) TBIL; (d) ALB; (e) CHE; (f) PT. Control: negative control group; Model: the group injected with D-GalN; SNMC: the group injected with D-GalN and treated with Stronger Neo-Minophagen C; Experiment: the group injected with D-GalN and treated with Qinggan Huoxue Recipe. For Control, Model, SNMC, and Experiment groups, the ALT levels were 35.15 ± 6.01 U/L, 441.10 ± 60.36 U/L, 267.18 ± 41.45 U/L, and 143.22 ± 22.96 U/L, respectively; the AST levels were 151.61 ± 20.87 U/L, 887.80 ± 128.47 U/L, 380.49 ± 55.38 U/L, and 287.36 ± 68.97 U/L, respectively; the TBIL levels were 1.55 ± 0.43 μmol/L, 38.04 ± 6.84 μmol/L, 24.37 ± 4.03 μmol/L, and 18.65 ± 2.96 μmol/L, respectively; the ALB levels were 35.25 ± 4.19 g/L, 23.67 ± 3.21 g/L, 26.65 ± 4.50 g/L, and 29.46 ± 4.19 g/L, respectively; the CHE levels were 557.40 ± 43.23 U/L, 343.92 ± 68.93 U/L, 430.50 ± 83.53 U/L, and 515.82 ± 73.31 U/L, respectively; and the PT was 13.60 ± 1.73 s, 31.80 ± 5.02 s, 29.93 ± 3.83 s, and 24.46 ± 4.25 s, respectively. For (a), (b), and (c), ^† P < 0.05 comparing Control group, *P < 0.05 comparing Model group, and ^‡ P < 0.05 comparing SNMC group. For (d), (e), ^& P < 0.05 comparing Control group and ^# P < 0.05 comparing Model group. For (f), ^& P < 0.05 comparing Control group and ^▲ P < 0.05 comparing Model group and SNMC group.

Figure 2

Pathological scores. (a) HE staining; (b) pathological scores. Control: negative control group; Model: the group injected with D-GalN; SNMC: the group injected with D-GalN and treated with Stronger Neo-Minophagen C; Experiment: the group injected with D-GalN and treated with Qinggan Huoxue Recipe. The pathological scores were 0, 3.69 ± 0.38, 2.78 ± 0.31, and 1.72 ± 0.21, respectively, in Control, Model, SNMC, and Experiment groups. ^& P < 0.05 comparing Control group, ^# P < 0.05 comparing Model group, and ^▲ P < 0.05 comparing SNMC group.

Figure 3

PCNA immunohistochemistry. (a) PCAN immunohistochemistry; (b) histogram of immunohistochemistry. Control: negative control group; Model: the group injected with D-GalN; SNMC: the group injected with D-GalN and treated with Stronger Neo-Minophagen C; Experiment: the group injected with D-GalN and treated with Qinggan Huoxue Recipe. The PCNA positive rates were 7.48 ± 0.90%, 17.55 ± 2.4%, 25.57 ± 2.94%, and 35.68 ± 4.75%, respectively, in Control, Model, SNMC, and Experiment groups. ^& P < 0.05 comparing Control group, ^# P < 0.05 comparing Model group, and ^▲ P < 0.05 comparing SNMC group.

Figure 4

The mRNA expression. (a) HMGB1; (b) TLR4; (c) NF-κB; (d) Caspase-3. Control: negative control group; Model: the group injected with D-GalN; SNMC: the group injected with D-GalN and treated with Stronger Neo-Minophagen C; Experiment: the group injected with D-GalN and treated with Qinggan Huoxue Recipe. For Control, Model, SNMC, and Experiment groups, the HMGB1 mRNA expressions were 0.01 ± 0.01, 0.25 ± 0.04, 0.11 ± 0.04, and 0.07 ± 0.06, respectively; the TLR4 mRNA expressions were 0.04 ± 0.02, 0.41 ± 0.22, 0.22 ± 0.08, and 0.08 ± 0.04, respectively; the NF-κB mRNA expressions were 0.49 ± 0.25, 2.68 ± 1.35, 1.78 ± 0.64, and 0.98 ± 0.63, respectively; and the Caspase-3 mRNA expressions were 1.36 ± 0.26, 3.41 ± 0.85, 2.57 ± 1.04, and 1.64 ± 0.81, respectively. ^& P < 0.05 comparing Control group, ^# P < 0.05 comparing Model group, and ^▲ P < 0.05 comparing SNMC group.

Figure 5

The survival curve. Model: the group injected with D-GalN; SNMC: the group injected with D-GalN and treated with Stronger Neo-Minophagen C; Experiment: the group injected with D-GalN and treated with Qinggan Huoxue Recipe. ^# P < 0.05 comparing Model group.