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Nrf2 pathway activation contributes to anti-fibrosis effects of ginsenoside Rg1 in a rat model of alcohol- and CCl4-induced hepatic fibrosis.

Li JP, Gao Y, Chu SF, Zhang Z, Xia CY, Mou Z, Song XY, He WB, Guo XF, Chen NH - Acta Pharmacol. Sin. (2014)

Bottom Line: Rg1 significantly increased the activities of antioxidant enzymes (SOD, GSH-Px and CAT) and reduced MDA levels in liver tissues.Furthermore, Rg1 significantly increased the expression and nuclear translocation of Nrf2 that regulated the expression of many antioxidant enzymes.Knockdown of Nrf2 gene diminished these actions of Rg1 in CCl4-treated HSCs in vitro.

View Article: PubMed Central - PubMed

Affiliation: State Key of Laboratory Bioactive Substances and Functions of Natural Medicines, Department of Pharmacology, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.

ABSTRACT

Aim: To investigate the anti-fibrosis effects of ginsenoside Rg1 on alcohol- and CCl4-induced hepatic fibrosis in rats and to explore the mechanisms of the effects.

Methods: Rats were given 6% alcohol in water and injected with CCl4 (2 mL/kg, sc) twice a week for 8 weeks. Rg1 (10, 20 and 40 mg/kg per day, po) was administered in the last 2 weeks. Hepatic fibrosis was determined by measuring serum biochemical parameters, HE staining, Masson's trichromic staining, and hydroxyproline and α-SMA immunohistochemical staining of liver tissues. The activities of antioxidant enzymes, lipid peroxidation, and Nrf2 signaling pathway-related proteins (Nrf2, Ho-1 and Nqo1) in liver tissues were analyzed. Cultured hepatic stellate cells (HSCs) of rats were prepared for in vitro studies.

Results: In the alcohol- and CCl4-treated rats, Rg1 administration dose-dependently suppressed the marked increases of serum ALT, AST, LDH and ALP levels, inhibited liver inflammation and HSC activation and reduced liver fibrosis scores. Rg1 significantly increased the activities of antioxidant enzymes (SOD, GSH-Px and CAT) and reduced MDA levels in liver tissues. Furthermore, Rg1 significantly increased the expression and nuclear translocation of Nrf2 that regulated the expression of many antioxidant enzymes. Treatment of the cultured HSCs with Rg1 (1 μmol/L) induced Nrf2 translocation, and suppressed CCl4-induced cell proliferation, reversed CCl4- induced changes in MDA, GPX, PCIII and HA contents in the supernatant fluid and α-SMA expression in the cells. Knockdown of Nrf2 gene diminished these actions of Rg1 in CCl4-treated HSCs in vitro.

Conclusion: Rg1 exerts protective effects in a rat model of alcohol- and CCl4-induced hepatic fibrosis via promoting the nuclear translocation of Nrf2 and expression of antioxidant enzymes.

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Nrf2 playing a key role in the mechanism of anti-hepatic fibrosis by Rg1. (A) Effect of Rg1 on prolieration in HSCs. Primary HSCs were incubated with Rg1 and CCl4 (10 mmol/L) for 24 h. Data represent mean±SD of 6 separate experiments. (B) Nrf2 siRNA inhibits Rg1-induced translocation of Nrf2. Cells were treated with 1 μmol/L Rg1 for 24 h after control or Nrf2 siRNA transfection for 48 h. Protein expression of Nrf2 was detected by Western blotting. Representative blots were from three independent experiments. The content of MDA (C), GPX (D), PCIII (E), and HA (F) in the cell cultured supernatant and the α-SMA expression in HSCs (G). Cells were treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h after control or Nrf2 siRNA transfection for 48 h; non-siRNA groups were cultured for 48 h and then treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h. The data were expressed as mean±SD. n=6. Statistical evaluation was performed using t-test. bP<0.05, cP<0.01 vs control. eP<0.05, fP<0.01 vs CCl4 alone. hP<0.05, iP<0.01 vs Rg1+CCl4 group.
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fig7: Nrf2 playing a key role in the mechanism of anti-hepatic fibrosis by Rg1. (A) Effect of Rg1 on prolieration in HSCs. Primary HSCs were incubated with Rg1 and CCl4 (10 mmol/L) for 24 h. Data represent mean±SD of 6 separate experiments. (B) Nrf2 siRNA inhibits Rg1-induced translocation of Nrf2. Cells were treated with 1 μmol/L Rg1 for 24 h after control or Nrf2 siRNA transfection for 48 h. Protein expression of Nrf2 was detected by Western blotting. Representative blots were from three independent experiments. The content of MDA (C), GPX (D), PCIII (E), and HA (F) in the cell cultured supernatant and the α-SMA expression in HSCs (G). Cells were treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h after control or Nrf2 siRNA transfection for 48 h; non-siRNA groups were cultured for 48 h and then treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h. The data were expressed as mean±SD. n=6. Statistical evaluation was performed using t-test. bP<0.05, cP<0.01 vs control. eP<0.05, fP<0.01 vs CCl4 alone. hP<0.05, iP<0.01 vs Rg1+CCl4 group.

Mentions: The CCl4-induced HSC proliferation was inhibited by Rg1 (10−7, 10−6 mol/L) compared with that of the CCl4 only group (P<0.05, P<0.01, respectively; Figure 7A). As 10−6 mol/L of Rg1 showed a better and more stable effect, this concentration was chosen for the following study.


Nrf2 pathway activation contributes to anti-fibrosis effects of ginsenoside Rg1 in a rat model of alcohol- and CCl4-induced hepatic fibrosis.

Li JP, Gao Y, Chu SF, Zhang Z, Xia CY, Mou Z, Song XY, He WB, Guo XF, Chen NH - Acta Pharmacol. Sin. (2014)

Nrf2 playing a key role in the mechanism of anti-hepatic fibrosis by Rg1. (A) Effect of Rg1 on prolieration in HSCs. Primary HSCs were incubated with Rg1 and CCl4 (10 mmol/L) for 24 h. Data represent mean±SD of 6 separate experiments. (B) Nrf2 siRNA inhibits Rg1-induced translocation of Nrf2. Cells were treated with 1 μmol/L Rg1 for 24 h after control or Nrf2 siRNA transfection for 48 h. Protein expression of Nrf2 was detected by Western blotting. Representative blots were from three independent experiments. The content of MDA (C), GPX (D), PCIII (E), and HA (F) in the cell cultured supernatant and the α-SMA expression in HSCs (G). Cells were treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h after control or Nrf2 siRNA transfection for 48 h; non-siRNA groups were cultured for 48 h and then treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h. The data were expressed as mean±SD. n=6. Statistical evaluation was performed using t-test. bP<0.05, cP<0.01 vs control. eP<0.05, fP<0.01 vs CCl4 alone. hP<0.05, iP<0.01 vs Rg1+CCl4 group.
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fig7: Nrf2 playing a key role in the mechanism of anti-hepatic fibrosis by Rg1. (A) Effect of Rg1 on prolieration in HSCs. Primary HSCs were incubated with Rg1 and CCl4 (10 mmol/L) for 24 h. Data represent mean±SD of 6 separate experiments. (B) Nrf2 siRNA inhibits Rg1-induced translocation of Nrf2. Cells were treated with 1 μmol/L Rg1 for 24 h after control or Nrf2 siRNA transfection for 48 h. Protein expression of Nrf2 was detected by Western blotting. Representative blots were from three independent experiments. The content of MDA (C), GPX (D), PCIII (E), and HA (F) in the cell cultured supernatant and the α-SMA expression in HSCs (G). Cells were treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h after control or Nrf2 siRNA transfection for 48 h; non-siRNA groups were cultured for 48 h and then treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h. The data were expressed as mean±SD. n=6. Statistical evaluation was performed using t-test. bP<0.05, cP<0.01 vs control. eP<0.05, fP<0.01 vs CCl4 alone. hP<0.05, iP<0.01 vs Rg1+CCl4 group.
Mentions: The CCl4-induced HSC proliferation was inhibited by Rg1 (10−7, 10−6 mol/L) compared with that of the CCl4 only group (P<0.05, P<0.01, respectively; Figure 7A). As 10−6 mol/L of Rg1 showed a better and more stable effect, this concentration was chosen for the following study.

Bottom Line: Rg1 significantly increased the activities of antioxidant enzymes (SOD, GSH-Px and CAT) and reduced MDA levels in liver tissues.Furthermore, Rg1 significantly increased the expression and nuclear translocation of Nrf2 that regulated the expression of many antioxidant enzymes.Knockdown of Nrf2 gene diminished these actions of Rg1 in CCl4-treated HSCs in vitro.

View Article: PubMed Central - PubMed

Affiliation: State Key of Laboratory Bioactive Substances and Functions of Natural Medicines, Department of Pharmacology, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.

ABSTRACT

Aim: To investigate the anti-fibrosis effects of ginsenoside Rg1 on alcohol- and CCl4-induced hepatic fibrosis in rats and to explore the mechanisms of the effects.

Methods: Rats were given 6% alcohol in water and injected with CCl4 (2 mL/kg, sc) twice a week for 8 weeks. Rg1 (10, 20 and 40 mg/kg per day, po) was administered in the last 2 weeks. Hepatic fibrosis was determined by measuring serum biochemical parameters, HE staining, Masson's trichromic staining, and hydroxyproline and α-SMA immunohistochemical staining of liver tissues. The activities of antioxidant enzymes, lipid peroxidation, and Nrf2 signaling pathway-related proteins (Nrf2, Ho-1 and Nqo1) in liver tissues were analyzed. Cultured hepatic stellate cells (HSCs) of rats were prepared for in vitro studies.

Results: In the alcohol- and CCl4-treated rats, Rg1 administration dose-dependently suppressed the marked increases of serum ALT, AST, LDH and ALP levels, inhibited liver inflammation and HSC activation and reduced liver fibrosis scores. Rg1 significantly increased the activities of antioxidant enzymes (SOD, GSH-Px and CAT) and reduced MDA levels in liver tissues. Furthermore, Rg1 significantly increased the expression and nuclear translocation of Nrf2 that regulated the expression of many antioxidant enzymes. Treatment of the cultured HSCs with Rg1 (1 μmol/L) induced Nrf2 translocation, and suppressed CCl4-induced cell proliferation, reversed CCl4- induced changes in MDA, GPX, PCIII and HA contents in the supernatant fluid and α-SMA expression in the cells. Knockdown of Nrf2 gene diminished these actions of Rg1 in CCl4-treated HSCs in vitro.

Conclusion: Rg1 exerts protective effects in a rat model of alcohol- and CCl4-induced hepatic fibrosis via promoting the nuclear translocation of Nrf2 and expression of antioxidant enzymes.

Show MeSH
Related in: MedlinePlus