Nutr Res Pract.  2020 Aug;14(4):334-351. 10.4162/nrp.2020.14.4.334.

Antioxidant and hepatoprotective effects of Korean ginseng extract GS-KG9 in a D-galactosamine-induced liver damage animal model

Affiliations
  • 1nternational Ginseng & Herb Research Institute, Geumsan 32724, Korea

Abstract

BACKGROUND/OBJECTIVES
This study was designed to investigate the improvement effect of white ginseng extract (GS-KG9) on D-galactosamine (Ga1N)-induced oxidative stress and liver injury.
SUBJECTS/METHODS
Sixty Sprague-Dawley rats were divided into 6 groups. Rats were orally administrated with GS-KG9 (300, 500, or 700 mg/kg) or silymarin (25 mg/kg) for 2 weeks. The rats of the GS-KG9- and silymarin-treated groups and a control group were then intraperitoneally injected Ga1N at a concentration of 650 mg/kg for 4 days. To investigate the protective effect of GS-KG9 against GalN-induced liver injury, blood liver function indicators, anti-oxidative stress indicators, and histopathological features were analyzed.
RESULTS
Serum biochemical analysis indicated that GS-KG9 ameliorated the elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) in GalN-treated rats. The hepatoprotective effects of GS-KG9 involved enhancing components of the hepatic antioxidant defense system, including glutathione, glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT). In addition, GS-KG9 treatment inhibited reactive oxygen species (ROS) production induced by GalN treatment in hepatocytes and significantly increased the expression levels of nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins, which are antioxidant proteins. In particular, by histological analyses bases on hematoxylin and eosin, Masson's trichrome, α-smooth muscle actin, and transforming growth factor-β1 staining, we determined that the administration of 500 mg/kg GS-KG9 inhibited hepatic inflammation and fibrosis due to the excessive accumulation of collagen.
CONCLUSIONS
These findings demonstrate that GS-KG9 improves GalN-induced liver inflammation, necrosis, and fibrosis by attenuating oxidative stress. Therefore, GS-KG9 may be considered a useful candidate in the development of a natural preventive agent against liver injury.

Keyword

Panax ginseng; liver; galactosamine; oxidative stress; liver fibrosis

Figure

  • Fig. 1 Schematic diagram of the experimental protocol. Rats were acclimatized on a normal diet for 1 week and then they were divided into 6 groups of ten animals each. They were orally administered the allocated samples once a day for 2 weeks. Each rat was intraperitoneally administered D-galactosamine 650 mg/kg and then all animals were sacrificed for further analysis on day 4 after D-galactosamine treatment.

  • Fig. 2 Effects of GS-KG9 on serum hepatic enzyme activities of rats with D-galactosamin-induced liver damage: (A) serum ALT, (B) serum AST, (C) serum ALP, and (D) serum LDH. Data are represented as the mean ± SE (n = 10). Values with different superscript letters are significantly different among groups according to a one-way analysis of variance coupled with Dunnett's multiple-range test (P < 0.05).ALT, alanine aminotransferase; AST, aspartate aminotransferase; ALP, alkaline phosphatase; LDH, lactate dehydrogenase; GalN, D-galactosamine-induced liver damage group; Sily, Silymarin 25 mg/kg treated and GalN-induced liver damage group; GS-300, GS-KG9 300 mg/kg treated and GalN-induced liver damage group; GS-500, GS-KG9 500 mg/kg treated and GalN-induced liver damage group; GS-700, GS-KG9 700 mg/kg treated and GalN-induced liver damage group.

  • Fig. 3 Effects of GS-KG9 on liver antioxidant enzyme activities of rats with D-galactosamin-induced liver damage. (A) SOD activity, (B) CAT activity, (C) GPX activity. Data are represented as the mean ± SE (n = 10). Values with different superscript letters are significantly different among groups according to a one-way analysis of variance coupled with Dunnett's multiple-range test (P < 0.05).SOD, superoxide dismutase; CAT, catalase; GPX, glutathione peroxidase; GalN, D-galactosamine-induced liver damage group; Sily, Silymarin 25 mg/kg treated and GalN-induced liver damage group; GS-300, GS-KG9 300 mg/kg treated and GalN-induced liver damage group; GS-500, GS-KG9 500 mg/kg treated and GalN-induced liver damage group; GS-700, GS-KG9 700 mg/kg treated and GalN-induced liver damage group.

  • Fig. 4 Inhibitory effects of GS-KG9 on ROS production in hepatocyte HapG2 cells. (A) cell viability of GS-KG9 in HapG2 cells, (B) cell viability of D-galactosamine in HapG2 cells, (C) ROS production induced by D-galactosamine in HapG2 cells, (D) Inhibitory effects of GS-KG9 on D-galactosamine-induced ROS production in HapG2 cells. Data are represented as the mean ± SE (n = 5). Values with different superscript letters are significantly different among groups according to a one-way analysis of variance coupled with Dunnett's multiple-range test (P < 0.05).ROS, reactive oxygen species; GalN, D-galactosamine.

  • Fig. 5 Effects of GS-KG9 on Nrf2 and HO-1 expression in GalN-treated HepG2 cells.GalN, D-galactosamine; Nucl-Nrf2, nuclear factor erythroid-2-related factor 2; HO-1, hemo oxygenase-1.

  • Fig. 6 Pathological examination of effects of GS-KG9 on liver inflammation in rats with GalN-induced liver injury. Livers were stained with hematoxylin and eosin and visualized at 100× magnification. (A) Normal group, (B) GalN-induced liver damage group, (C) Silymarin 25 mg/kg treated and GalN-induced liver damage group, (D) GS-KG9 300 mg/kg treated and GalN-induced liver damage group, (E) GS-KG9 500 mg/kg treated and GalN-induced liver damage group, and (F) GS-KG9 700 mg/kg treated and GalN-induced liver damage group.GalN, D-galactosamine.

  • Fig. 7 Pathological examination of effects of GS-KG9 on liver fibrosis in rats with GalN-induced liver injury. Livers were stained with Masson's trichrome and visualized at 100× magnification. (A) Normal group, (B) GalN-induced liver damage group, (C) Silymarin 25 mg/kg treated and GalN-induced liver damage group, (D) GS-KG9 300 mg/kg treated and GalN-induced liver damage group, (E) GS-KG9 500 mg/kg treated and GalN-induced liver damage group, and (F) GS-KG9 700 mg/kg treated and GalN-induced liver damage group.GalN, D-galactosamine.

  • Fig. 8 Effects of GS-KG9 on activation of hepatic stellate cells in rats with GalN-induced liver injury. Immunohistochemical staining of α-smooth muscle actin was performed on liver sections and visualized at 100× magnification. (A) Normal group, (B) GalN-induced liver damage group, (C) Silymarin 25 mg/kg treated and GalN-induced liver damage group, (D) GS-KG9 300 mg/kg treated and GalN-induced liver damage group, (E) GS-KG9 500 mg/kg treated and GalN-induced liver damage group, and (F) GS-KG9 700 mg/kg treated and GalN-induced liver damage group.GalN, D-galactosamine.

  • Fig. 9 Effects of GS-KG9 on TGF-β1 stain expression in rats with GalN-induced liver injury. Immunohistochemical staining of TGF-β1 was performed on liver sections and visualized at 100× magnification. (A) Normal group, (B) GalN-induced liver damage group, (C) Silymarin 25 mg/kg treated and GalN-induced liver damage group, (D) GS-KG9 300 mg/kg treated and GalN-induced liver damage group, (E) GS-KG9 500 mg/kg treated and GalN-induced liver damage group, and (F) GS-KG9 700 mg/kg treated and GalN-induced liver damage group.TGF-β1, transforming growth factor-β1; GalN, D-galactosamine.


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