Diabetes Metab J.  2011 Oct;35(5):489-496. 10.4093/dmj.2011.35.5.489.

Dietary Oleate Has Beneficial Effects on Every Step of Non-Alcoholic Fatty Liver Disease Progression in a Methionine- and Choline-Deficient Diet-Fed Animal Model

• Lee JY
• Moon JH
• Park JS
• Lee BW
• Kang ES
• Ahn CW
• Lee HC
• Cha BS

• KMID: 1857504
• DOI: http://doi.org/10.4093/dmj.2011.35.5.489

Abstract

BACKGROUND
Non-alcoholic fatty liver disease (NAFLD) is increasingly recognized as a major cause of liver-related morbidity and mortality. The underlying mechanisms of disease progression remain poorly understood, and primary therapy of NAFLD is not yet established. We investigated the effects of dietary oleate on the development and progression of NAFLD in a methionine- and choline-deficient (MCD) diet-fed animal model.
METHODS
A total of 30 C57BL/6J mice were randomly divided into three groups (n=10 in each group) and fed various experimental diets for four weeks: chow, MCD diet, or OMCD (MCD diet with oleate, 0.5 mg/g/day). Liver samples were examined for steatohepatitis and fibrosis parameters and associated genes.
RESULTS
Additional dietary oleate dramatically reduced MCD diet-induced hepatic steatosis. Hepatic carbohydrate responsive element-binding protein was overexpressed in MCD diet-fed mice, and dietary oleate prevented this overexpression (P<0.001). Dietary oleate partially prevented MCD diet-induced serum level increases in aspartate aminotransferase and alanine aminotransferase (P<0.001, respectively). The mRNA expressions of hepatic monocyte chemoattractant protein 1, tumor necrosis factor-alpha and matrix metalloproteinase-9 were increased in MCD diet-fed mice, and this overexpression of inflammatory molecules was prevented by dietary oleate (P<0.001). Hepatic pericellular fibrosis was observed in MCD diet-fed mice, and dietary oleate prevented this fibrosis. Altogether, dietary oleate prevented MCD diet-induced hepatic steatosis, inflammation and fibrosis.
CONCLUSION
Dietary oleate has beneficial effects in every step of NAFLD development and progression and could be a nutritional option for NAFLD prevention and treatment.

Keyword

ChREBP; Fatty acids, monounsaturated; Non-alcoholic steatohepatitis

MeSH Terms

Alanine Transaminase
Animals
Aspartate Aminotransferases
Chemokine CCL2
Corneal Dystrophies, Hereditary
Diet
Disease Progression
Fatty Acids, Monounsaturated
Fatty Liver
Fibrosis
Inflammation
Liver
Matrix Metalloproteinase 9
Mice
Models, Animal
Oleic Acid
RNA, Messenger
Tumor Necrosis Factor-alpha
Alanine Transaminase
Aspartate Aminotransferases
Chemokine CCL2
Corneal Dystrophies, Hereditary
Fatty Acids, Monounsaturated
Fatty Liver
Matrix Metalloproteinase 9
Oleic Acid
RNA, Messenger
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1 The effects of the methionine- and choline-deficient diet (MCD) diet and dietary oleate on hepatic fat accumulation. Oil-red-O staining of frozen liver sections. Scale bars, 200 µm. (A) Chow diet-fed mice. (B) MCD diet-fed mice. (C) MCD diet with oleate (0.5 mg/g/day for 4 weeks)-fed mice. (D) The areas of fat droplets, measured by image analyzer in three randomly selected fields (magnification, ×100) of each liver section. Chow, normal chow diet; OMCD, MCD diet with oleate. aP<0.05 vs. chow, bP

• 

  Fig. 2 The effects of the methionine- and choline-deficient diet (MCD) diet and dietary oleate on hepatic carbohydrate responsive element-binding protein (ChREBP) and apolipoprotein B (apo B) expression. Immunohistochemical detection of ChREBP in liver paraffin-embedded sections. Scale bars, 50 µm. (A) Chow diet-fed mice. (B) MCD diet-fed mice. (C) MCD diet with oleate (0.5 mg/g/day for 4 weeks)-fed mice. (D) Real-time reverse transcription-polymerase chain reaction (RT-PCR) quantification of ChREBP mRNA in mouse liver samples from each group. (E) Real-time RT-PCR quantification of APOB100 mRNA in mouse liver samples from each group. AU, arbitrary unit; Chow, normal chow diet; OMCD, MCD diet with oleate. aP<0.05 vs. chow, bP

• 

  Fig. 3 Liver histology in mice fed each experimental diet. H&E staining of liver paraffin-embedded sections. Scale bars, 200 µm. (A) Chow diet-fed mice. (B) MCD diet-fed mice. (C) MCD diet with oleate (0.5 mg/g/day for 4 weeks)-fed mice. Arrows indicate the focal aggregation of inflammatory cells. Chow, normal chow diet; MCD, methionine- and choline-deficient diet; OMCD, MCD diet with oleate.

• 

  Fig. 4 The effects of the methionine- and choline-deficient diet (MCD) diet and dietary oleate on hepatic inflammatory molecules. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) for inflammatory molecules in mouse liver samples from each group. (A) MCP1. (B) TNF-α. (C) MMP-9. MCP1, monocyte chemotactic protein 1; TNF-α, tumor necrosis factor-α; MMP-9, matrix metalloproteinase-9; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; AU, arbitrary unit; Chow, normal chow diet; MCD, methionine- and choline-deficient diet; OMCD, MCD diet with oleate (0.5 mg/g/day for 4 weeks). aP<0.05 vs. chow, bP

• 

  Fig. 5 The effects of the methionine- and choline-deficient diet (MCD) diet and dietary oleate on hepatic fibrosis. Trichrome staining of liver paraffin-embedded sections. Scale bars, 200 µm. (A) Chow diet-fed mice. (B) MCD diet-fed mice. (C) MCD diet with oleate (0.5 mg/g/day for 4 weeks)-fed mice. (D) The areas of fibrosis, measured by image analyzer in three randomly selected fields (magnification, ×100) of each liver section. AU, arbitrary unit; Chow, normal chow diet; OMCD, MCD diet with oleate. aP<0.05 vs. chow, bP

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