Nutr Res Pract.  2018 Apr;12(2):110-117. 10.4162/nrp.2018.12.2.110.

Ethanol extract of Allium fistulosum inhibits development of non-alcoholic fatty liver disease

• Hwang JT
• Shin EJ
• Chung MY
• Park JH
• Chung S
• Choi HK

• KMID: 2407807
• DOI: http://doi.org/10.4162/nrp.2018.12.2.110

Abstract

BACKGROUND/OBJECTIVES
Non-alcoholic fatty liver disease (NAFLD) is a leading cause of chronic liver disease and is closely associated with metabolic syndrome. In the present study, we observed the effect of ethanol extract of Allium fistulosum (EAF) on NAFLD and have suggested the possibility of using EAF as a natural product for application in the development of a treatment for NAFLD.
MATERIALS/METHODS
The preventive effect on hepatic lipid accumulation was estimated by using an oleic acid (OA)-induced NAFLD model in vitro and a Western diet (high-fat high-sucrose; WD)-induced obese mouse model. Animals were divided into three groups (n = 7): normal diet group (ND), WD group, and WD plus 1% EAF group.
RESULTS
EAF reduced OA-stimulated lipid accumulation in HepG2 cells in the absence of cellular cytotoxicity and significantly blocked transcriptional activation of sterol regulatory element-binding protein 1 and fatty acid synthase genes. Subsequently, we investigated these effects in vivo in mice fed either ND or WD in the presence or absence of EAF supplementation. In comparison to the ND controls, the WD-fed mice exhibited increases in body weight, liver weight, epididymal fat weight, and accumulation of fat in hepatocytes, and these effects were significantly attenuated by EAF supplementation.
CONCLUSIONS
Allium fistulosum attenuates the development of NAFLD, and EAF elicits anti-lipogenic activity in liver. Therefore, EAF represents a promising candidate for use in the development of novel therapeutic drugs or drug combinations for the prevention and treatment of NAFLD.

Keyword

Non-alcoholic fatty liver disease (NAFLD); metabolic syndrome; western diet; lipogenesis; HepG2 cells

MeSH Terms

Allium*
Animals
Body Weight
Diet
Diet, Western
Drug Combinations
Ethanol*
Hep G2 Cells
Hepatocytes
In Vitro Techniques
Lipogenesis
Liver
Liver Diseases
Mice
Mice, Obese
Non-alcoholic Fatty Liver Disease*
Oleic Acid
Sterol Regulatory Element Binding Protein 1
Transcriptional Activation
Drug Combinations
Ethanol
Oleic Acid
Sterol Regulatory Element Binding Protein 1

Figure

• 

  Fig. 1 Ethanol extract of Allium fistulosum (EAF) attenuates lipid accumulation in HepG2 cells. A) Non-cytotoxic concentrations of EAF significantly block oleic acid (OA)-mediated lipid accumulation. HepG2 cells treated in the presence or absence of OA, alone or in combination with EAF, were stained by using an Oil Red O solution. The values presented are mean ± SD for three independent experiments. Means not sharing a common superscript are significantly different, P < 0.05. (B) EAF at concentrations up to 400 µg/mL did not produce cytotoxicity in HepG2 cells. Cell viability was measured by using a WST-1 assay in HepG2 cells treated in the presence or absence of OA, alone or in combination with EAF, for 24 h. There was no statistical significance detected.

• 

  Fig. 2 Ethanol extract of Allium fistulosum (EAF) inhibits transcriptional activity of lipogenesis-related genes. The mRNA expressions of genes associated with lipid metabolism were measured by performing qRT-PCR. EAF was treated alone or in combination at either 100 or 200 µg/mL in the presence or absence of oleic acid (OA) in HepG2 cells for 18 h. All experiments were conducted independently three times and data are expressed as mean ± SD. Means not sharing a common superscript are significantly different, P < 0.05. SREBP1c, Sterol regulatory element-binding protein 1; FASN, fatty acid synthase; ACLY, ATP citrate lyase.

• 

  Fig. 3 Body weight, daily food intake, and food efficiency ratio analysis in mice fed on a Western diet (WD) supplemented in the presence or absence of 1% ethanol extract of Allium fistulosum (EAF) for 12 weeks. (A) WD-fed mice exhibit increased body weight compared to the controls, while 1% EAF supplementation significantly reduced body weight. Data are expressed as mean ± SE (n = 7/group). (B) The average weight gain after 12-week supplementation is expressed as mean ± SE (n = 7/group). (C) Daily food intake (g) in the WD-fed group is lower than in their control counterparts, but there are no significant differences among the three groups. The data are expressed as mean ± SE (n = 7/group). (D) The increased food efficiency ratio in WD-fed group was reduced by 1% EAF supplementation. Food efficiency ratio (FER) was calculated by applying the equation: FER = (body weight gain (g)/food intake (g)) × 100. FER is expressed as mean ± SE (n = 7/group). Means not sharing a common superscript are significantly different, P < 0.05.

• 

  Fig. 4 The 1% ethanol extract of Allium fistulosum (EAF) supplement blocked gains in both liver and epididymal fat weights and attenuated hepatic lipid accumulation. (A) Liver weight was significantly increased in Western diet (WD)-fed mice and was rapidly reduced by 1% EAF supplementation with WD. The data are expressed as mean ± SE (n = 7/group). (B) Mice fed with WD exhibit greater lipid accumulation compared to the controls, while EAF supplementation markedly attenuated this effect. Representative images are shown with a 100 µm scale bar. (C) The epididymal fat weight was significantly increased in WD-fed mice and was reduced by 1% EAF supplementation. The data are expressed as mean ± SE (n = 7/group). Means not sharing a common superscript are significantly different, P < 0.05.

• 

  Fig. 5 The 1% ethanol extract of Allium fistulosum (EAF) supplement markedly reduced Western diet (WD)-induced liver abnormalities. (A) Plasma levels of alanine aminotransferase (ALT) were dramatically increased in WD-fed mice but were reduced by 1% EAF supplementation. Values are mean ± SE (n = 7/group). (B) Plasma levels of aspartate transaminase (AST) were statistically increased in WD-fed mice, and 1% EAF supplementation reduced that level; however, there was no statistical significance to the changes. Values are mean ± SE (n = 7/group). (C) The reduced AST/ALT ratio in WD-fed group was dramatically recovered by 1% EAF supplementation. The data are expressed as mean ± SE (n = 7/group). (D) Plasma levels of ALP were significantly increased in WD-fed mice and were reduced by 1% EAF supplementation. Values are mean ± SE (n = 7/group). Means not sharing a common superscript are significantly different, P < 0.05.

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