Efficacy of Boesenbergia rotunda Treatment against Thioacetamide-Induced Liver Cirrhosis in a Rat Model

Abstract

Background. Experimental research in hepatology has focused on developing traditional medicines into potential pharmacological solutions aimed at protecting liver from cirrhosis. Along the same line, this study investigated the effects of ethanol-based extract from a traditional medicine plant Boesenbergia rotunda (BR) on liver cirrhosis. Methodology/Results. The BR extract was tested for toxicity on 3 groups of rats subjected to vehicle (10% Tween 20, 5 mL/kg) and 2g/kg and 5g/kg doses of the extract, respectively. Next, experiments were conducted on a rat model of cirrhosis induced by thioacetamide injection. The rats were divided into five groups and, respectively, administered orally with 10% Tween-20 (5 mL/kg) (normal control group), 10% Tween-20 (5 mL/kg) (cirrhosis control group), 50 mg/kg of silymarin (reference control group), and 250 mg/kg and 500 mg/kg of BR extract (experimental groups) daily for 8 weeks. The rats in normal group were intraperitoneally injected with sterile distilled water (1 mL/kg) 3 times/week, and those in the remaining groups were injected intraperitoneally with thioacetamide (200 mg/kg) thrice weekly. At the end of the 8 weeks, the animals were sacrificed and samples were collected for comprehensive histopathological, coagulation profile and biochemical evaluations. Also, the antioxidant activity of the BR extract was determined and compared with that of silymarin. Data from the acute toxicity tests showed that the extract was safe to use. Histological analysis of the livers of the rats in cirrhosis control group revealed uniform coarse granules on their surfaces, hepatocytic necrosis, and lymphocytes infiltration. But, the surfaces morphologically looked much smoother and the cell damage was much lesser in those livers from the normal control, silymarin and BR-treated groups. In the high-dose BR treatment group, the livers of the rats exhibited nearly normal looking lobular architecture, minimal inflammation, and minimal hepatocyte damage, the levels of the serum biomarkers and liver enzymes read nearly normal, and these results were all comparable to those observed or quantified from the normal and silymarin-treated groups. The BR extract had the antioxidant activity about half of what was recorded for silymarin. Conclusion. The progression of the liver cirrhosis can be intervened using the ethanol-based BR extract, and the liver's status quo of property, structure, and function can be preserved. This capability of the extract warrants further studies exploring the significance of its pharmacologic potential in successfully treating the liver cirrhosis in humans.

Figure 1

Antioxidant activity of the BR extract compared with the following standards: Gallic acid, Quercetin, Ascorbic acid, Rutin, Trolox, BHT as well as the standard drug Silymarin. Values were expressed as Mean ± SEM. Significant value was at P < 0.001.

Figure 2

Examples of H&E-stained histological sections of livers (left column) and kidneys (right column) obtained from the rats in the acute toxicity test. Rat with the liver and kidney shown in (a) and (b) was treated with 5 mL/kg vehicle (10% Tween 20). Rat with the liver and kidney shown in (c) and (d) was treated with 2 g/kg (5 mL/kg) dose of the BR extract. Rat with the liver and kidney shown in (e) and (f) was treated with 5 g/kg (5 mL/kg) dose of the BR extract. There was no significant difference in the structures of the liver and kidney between the treatment and control groups.

Figure 3

Example images showing macroscopic appearances of the livers sampled from rats in different experimental groups. (a) The liver of a control rat exhibiting regular smooth surface. (b) The liver of a hepatotoxic rat depicting numerous irregular whitish micro- and macronodular on its surface and a large area of ductular cholangiocellular proliferation (arrow) embedded within fibrosis. (c) The liver of a hepatoprotective rat treated with silymarin showing normal smooth surface. (d) The liver of a rat treated with 250 mg/kg of the BR extract illustrating nearly smooth surface with fewer granules (arrow head). (e) The liver of a rat treated with 500 mg/kg of the BR extract having normal smooth surface.

Figure 4

Example histopathological sections of livers sampled from rats in different experimental groups. (a) Normal histological structure and architecture were seen in livers of the control rats. (b) Severe structural damage, formation of pseudolobules with thick fibrotic septa and necrotic areas were present in the liver of the hepatotoxic rat. (c) Mild inflammation but no fibrotic septa was depicted in the liver of the hepatoprotective rat treated with silymarin. (d) Partially preserved hepatocyte and architecture with small area of necrosis and fibrotic septa existed in the liver of the rat treated with 250 mg/kg of the BR extract. (e) Partially preserved hepatocyte and architecture with small areas of mild necrosis were observed in the liver of the rat treated with 500 mg/kg of the BR extract. (H&E stain original magnification 20x).

Figure 5

Effect of BR ethanol extract on % cell normality. Data are expressed as mean ± SEM. Means among groups (n = 6 rate/group) show significant difference, *P < 0.001 compared to cirrhosis control group and **P < 0.001 compared to normal control group.

Figure 6

Effect of the BR extract on the level of MDA in the liver tissue. Data were expressed as Mean ± SEM. Means between the silymarin treated Group 3, low-dose BR-treated Group 4, and high-dose BR-treated Group 5 had significant differences when compared with the cirrhosis control Group 2 with *P < 0.001  and compared with the normal control Group 1 with **P < 0.001.

Figure 7

Effect of BR ethanol group on SOD level in the liver tissue. Data are expressed as mean ± SEM. Means among groups (n = 6 rate/group) show significant difference, *P < 0.001 compared to cirrhosis control group, and **P < 0.001 compared to normal control group.