J Vet Sci.  2016 Sep;17(3):279-287. 10.4142/jvs.2016.17.3.279.

Aristolochia manshuriensis Kom ethyl acetate extract protects against high-fat diet-induced non-alcoholic steatohepatitis by regulating kinase phosphorylation in mouse

• Kwak DH
• Kim JS
• Chang KT
• Choo YK

• KMID: 2413126
• DOI: http://doi.org/10.4142/jvs.2016.17.3.279

Abstract

Aristolochia manshuriensis Kom (AMK) is an herb used as a traditional medicine; however, it causes side effects such as nephrotoxicity and carcinogenicity. Nevertheless, AMK can be applied in specific ways medicinally, including via ingestion of low doses for short periods of time. Non-alcoholic steatohepatitis (NASH) induced the hepatocyte injury and inflammation. The protective effects of AMK against NASH are unclear; therefore, in this study, the protective effects of AMK ethyl acetate extract were investigated in a high-fat diet (HFD)-induced NASH model. We found decreased hepatic steatosis and inflammation, as well as increased levels of lipoproteins during AMK extract treatment. We also observed decreased hepatic lipid peroxidation and triglycerides, as well as suppressed hepatic expression of lipogenic genes in extract-treated livers. Treatment with extract decreased the activation of c-jun N-terminal kinase 1/2 (JNK1/2) and increased the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). These results demonstrate that the protective effect of the extract against HFD-induced NASH occurred via reductions in reactive oxygen species production, inflammation suppression, and apoptosis related to the suppression of JNK1/2 activation and increased ERK1/2 phosphorylation. Taken together, these results indicate that that ethyl acetate extract of AMK has potential therapeutic effects in the HFD-induced NASH mouse model.

Keyword

Aristolochia manshuriensis Kom; c-jun N-terminal kinase 1/2; extracellular signal-regulated kinase 1/2; inflammation; nonalcoholic steatohepatitis

MeSH Terms

Acetates/chemistry
Animals
Aristolochia/*chemistry
*Diet, High-Fat
Extracellular Signal-Regulated MAP Kinases/*genetics/metabolism
JNK Mitogen-Activated Protein Kinases/*genetics/metabolism
Liver/*drug effects/metabolism
Male
Mice
Mice, Inbred C57BL
Non-alcoholic Fatty Liver Disease/*drug therapy
Phosphorylation/drug effects
Plant Extracts/*pharmacology/*therapeutic use
Acetates
Plant Extracts
Extracellular Signal-Regulated MAP Kinases
JNK Mitogen-Activated Protein Kinases

Figure

• 

  Fig. 1 Effects of EtOAc extract of AMK on non-alcoholic steatohepatitis (NASH) in mice. (A) Morphological evaluation of fatty liver. (B) Evaluation of triglyceride (TG) and cholesterol (TC) in NASH mice sera. The levels were measured in mice fed with ND, HFD, or HFD plus EtOAc extract of AMK (2.5 mg/kg) for 15 weeks. Each group consisted of 10 mice. Data are expressed as the means ± SD. *p < 0.05, ***p < 0.001 vs. HFD-fed mice.

• 

  Fig. 2 Effects of EtOAc extract of AMK on NASH mice. (A) Serum levels of high-density lipoprotein (HDL), low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL) in NASH mice. (B) alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) in mice with NASH. The levels were measured in mice fed with ND, HFD, or HFD plus EtOAc extract of AMK (2.5 mg/kg) for 15 weeks. Each group consists of 10 mice. Data are expressed as the means ± SD. *p < 0.05 vs. HFD-fed mice.

• 

  Fig. 3 Effects of EtOAc extract of AMK on hepatic TG and lipoperoxide content in NASH mice. (A) Hepatic TG and (B) lipoperoxide content were assessed in mice fed ND, HFD, or HFD treated with EtOAc extract of AMK (2.5 mg/kg) for 12 weeks. Data are the means ± SD of 10 mice in each group. *p < 0.05 vs. HFD-fed mice. SOD, superoxide dismutase; GPx, glutathione peroxidase; GR, glutathione reductase; GSH, glutathione; TBARS, thiobarbituric acid reactive substances.

• 

  Fig. 4 Effects of HFD and treatment with EtOAc extract of AMK on hepatic mRNA expression. Expression of regulation genes such as (A) hepatic fatty acid, (B) adiopogenesis and (C) inflammation was measured in the hepatocytes of mice fed ND, HFD, or HFD plus EtOAc extract of AMK (2.5 mg/kg) for 15 weeks. Data are the means ± SD of 10 mice in each group. *p < 0.05, **p < 0.01 vs. HFD-fed mice.

• 

  Fig. 5 Effect of AMK-EtOAc extract on inflammatory and fatty acid regulation proteins. (A) Mitogen-activated protein kinases proteins including ERK1/2 and JNK1/2 in hepatocytes of mice with NASH. (B) TNF-α, CYP2E1, PPAR-α, Bax, caspase-3, ERK1/2, and JNK1/2 proteins in hepatocytes of mice with NASH. Data are the means ± SD of 10 mice in each group.

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