Nutr Res Pract.  2014 Dec;8(6):655-661. 10.4162/nrp.2014.8.6.655.

Arctiin inhibits adipogenesis in 3T3-L1 cells and decreases adiposity and body weight in mice fed a high-fat diet

• Min B
• Lee H
• Song JH
• Han MJ
• Chung J

• KMID: 2313792
• DOI: http://doi.org/10.4162/nrp.2014.8.6.655

Abstract

BACKGROUND/OBJECTIVES
The purpose of this study was to examine the effects and associated mechanisms of arctiin, a lignan compound found in burdock, on adipogenesis in 3T3-L1 cells. Also, the effects of arctiin supplementation in obese mice fed a high-fat diet on adiposity were examined.
MATERIALS/METHODS
3T3-L1 cells were treated with arctiin (12.5 to 100 microM) during differentiation for 8 days. The accumulation of lipid droplets was determined by Oil Red O staining and intracellular triglyceride contents. The expressions of genes related to adipogenesis were measured by real-time RT-PCR and Western blot analyses. For in vivo study, C57BL/6J mice were first fed either a control diet (CON) or high-fat diet (HF) to induce obesity, and then fed CON, HF, or HF with 500 mg/kg BW arctiin (HF + AC) for four weeks.
RESULTS
Arctiin treatment to 3T3-L1 pre-adipocytes markedly decreased adipogenesis in a dose-dependent manner. The arctiin treatment significantly decreased the protein levels of the key adipogenic regulators PPARgamma and C/EBPalpha, and also significantly inhibited the expression of SREBP-1c, fatty acid synthase, fatty acid-binding protein and lipoprotein lipase. Also, arctiin greatly increased the phosphorylation of AMP-activated protein kinase (AMPK) and its downstream target phosphorylated-acetyl CoA carboxylase. Furthermore, administration of arctiin significantly decreased the body weight in obese mice fed with the high-fat diet. The epididymal, perirenal or total visceral adipose tissue weights of mice were all significantly lower in the HF + AC than in the HF. Arctiin administration also decreased the sizes of lipid droplets in the epididymal adipose tissue.
CONCLUSIONS
Arctiin inhibited adipogenesis in 3T3-L1 adipocytes through the inhibition of PPARgamma and C/EBPalpha and the activation of AMPK signaling pathways. These findings suggest that arctiin has a potential benefit in preventing obesity.

Keyword

Arctiin; adipogenesis; AMP kinase; 3T3-L1 cells; high-fat diet

MeSH Terms

3T3-L1 Cells*
Adenylate Kinase
Adipocytes
Adipogenesis*
Adipose Tissue
Adiposity*
AMP-Activated Protein Kinases
Animals
Blotting, Western
Body Weight*
Diet
Diet, High-Fat*
Intra-Abdominal Fat
Lipoprotein Lipase
Mice*
Mice, Obese
Obesity
Phosphorylation
PPAR gamma
Sterol Regulatory Element Binding Protein 1
Triglycerides
Weights and Measures
AMP-Activated Protein Kinases
Adenylate Kinase
Lipoprotein Lipase
PPAR gamma
Sterol Regulatory Element Binding Protein 1

Figure

• 

  Fig. 1 Effects of arctiin on the differentiation and adipogenesis of 3T3-L1 cells. 3T3-L1 pre-adipocytes were incubated with MDI (DMEM with 3-isobutyl-1-methylxanthine, dexamethasone, and insulin) for 2 days and then replaced with DMEM containing insulin with or without arctiin (0, 12.5, 25, 50, and 100 µM) for 8 days. (A) Intracellular lipid droplets were stained with Oil Red O and observed at magnification 200 ×. (B) Intensities of Oil Red O staining measured by spectrophotometric analysis at 520 nm. (C) Intracellular triglyceride concentrations. Data are presented as the mean ± SE from three independent experiments. Different letters indicate significant difference (P < 0.05).

• 

  Fig. 2 Effects of arctiin treatment on cell viability in 3T3-L1 cells. 3T3-L1 pre-adipocytes were incubated with MDI (DMEM with 3-isobutyl-1-methylxanthine, dexamethasone, and insulin) for 2 days and then replaced with DMEM containing insulin with or without arctiin (0, 12.5, 25, 50, and 100 µM) for 8 days. Cell viability was determined by MTT assay. Data are presented as the mean ± SE from three independent experiments. Different letters indicate significant difference (P < 0.05).

• 

  Fig. 3 Effects of arctiin on the protein levels of adipogenic transcription factors in 3T3-L1 cells. The protein levels of PPARγ and C/EBPα in 3T3-L1 cells were determined by Western blot analyses. (A) Representative Western blot. (B) Densitometric analyses. Data are presented as the mean ± SE from three independent experiments. Different letters indicate significant difference (P < 0.05).

• 

  Fig. 4 Effects of arctiin on the expression of adipogenic genes in 3T3-L1 cells. The mRNA levels of adipogenic genes were examined by real-time RT-PCR. Data are presented as the mean ± SE from three independent experiments. Different letters indicate significant difference (P < 0.05).

• 

  Fig. 5 Effects of arctiin on AMPK phosphorylation in 3T3-L1 cells. The phosphorylation of AMPK and ACC in 3T3-L1 cells were determined by Western blot analyses. (A) Representative Western blot. Densitometric analyses for AMPK phosphorylation (B) and ACC phosphorylation (C) Data are presented as the mean ± SE from three independent experiments. Different letters indicate significant difference (P < 0.05).

• 

  Fig. 6 Effects of arctiin on the cell sizes of the epididymal adipose tissues in mice fed a high-fat diet. Hematoxylin and eosin-stained sections of epididymal adipose tissues are shown at 100 × magnification.

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