Nutr Res Pract.  2012 Aug;6(4):286-293.

Anti-obesity effect of resveratrol-amplified grape skin extracts on 3T3-L1 adipocytes differentiation

• Zhang XH
• Huang B
• Choi SK
• Seo JS

Abstract

Resveratrol (3,4,5-trihydroxy-trans-stilbene), a phytoalexin found in grape skin, grape products, and peanuts as well as red wine, has been reported to have various biological and pharmacological properties. The purpose of this study was to investigate the anti-obesity effect of resveratrol-amplified grape skin extracts on adipocytes. The anti-obesity effects of grape skin extracts were investigated by measuring proliferation and differentiation in 3T3-L1 cells. The effect of grape skin ethanol extracts on cell proliferation was detected by the MTS assay. The morphological changes and degree of adipogenesis of preadipocyte 3T3-L1 cells were measured by Oil Red-O staining assay. Treatment with extracts of resveratrol-amplified grape skin decreased lipid accumulation and glycerol-3-phosphate dehydrogenase activity without affecting 3T3-L1 cell viability. Grape skin extract treatment resulted in significantly attenuated expression of key adipogenic transcription factors, including peroxisome proliferator-activated receptor, CCAAT/enhancer-binding proteins, and their target genes (FAS, aP2, SCD-1, and LPL). These results indicate that resveratrol-amplified grape skin extracts may be useful for preventing obesity by regulating lipid metabolism.

Keyword

Grape skin extracts; resveratrol; 3T3-L1; adipocyte differentiation

MeSH Terms

3T3-L1 Cells
Adipocytes
Adipogenesis
Arachis
Cell Proliferation
Ethanol
Glycerolphosphate Dehydrogenase
Lipid Metabolism
Obesity
Peroxisomes
Proteins
Sesquiterpenes
Skin
Stilbenes
Transcription Factors
Vitis
Wine
Ethanol
Glycerolphosphate Dehydrogenase
Proteins
Sesquiterpenes
Stilbenes
Transcription Factors

Figure

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  Fig. 1 Effect of the 50% (A) and 80% (B) resveratrol-amplified grape skin extracts on preadipocyte viability. 3T3-L1 preadipocytes were incubated with resveratrol-amplified grape skin extracts at various concentrations (0, 30, 60, 120, 250, 500, and 1,000 µg/ml) for 24 and 48 h. Mean ± SD (n = 3).

• 

  Fig. 2 Effect of the resveratrol-amplified grape skin extracts on inhibiting 3T3-L1 adipocyte differentiation. 3T3-L1 cells differentiated with differentiation media in the absence or presence of resveratrol-amplified grape skin extract (RAGE, 200 and 400 µg/ml) for 8 days. (A) Intracellular lipids were stained with Oil Red-O. The results were confirmed by three independent experiments. (B) Stained triglyceride content was quantified by measuring absorbance. Non, undifferentiated cells; DM, differentiated media cells. Values are mean ± SD (n = 3). Test for DM vs. Non: †††P < 0.001. Test for RAGE vs. DM: **P < 0.01, ***P < 0.001.

• 

  Fig. 3 Effect of resveratrol-amplified grape skin extracts on glycerol-3-phosphate dehydrogenase (GPDH) activity during 3T3-L1 adipocyte differentiation. Confluent 3T3-L1 preadipocytes were differentiated into adipocytes in medium with or without different concentrations of resveratrol-amplified grape skin extracts (RAGE) for 8 days. Non, undifferentiated cells; DM, differentiated media cells. Values are mean ± SD (n = 3). Test for DM vs. Non: †††P < 0.001. Test for RAGE vs. DM: **P < 0.01, ***P < 0.001.

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  Fig. 4 Effect of resveratrol-amplified grape skin extracts on PPARγ and C/EBPα gene expression. Confluent 3T3-L1 preadipocytes were differentiated into adipocytes in medium with or without different concentrations of resveratrol-amplified grape skin extracts (RAGE) for 8 days. Non, undifferentiated cells; DM, differentiated media cells. Values are mean ± SD (n = 3). Test for DM vs. Non: †††P < 0.001. Test for RAGE vs. DM: **P < 0.01, ***P < 0.001.

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  Fig. 5 Effect of resveratrol-amplified grape skin extracts on PPARγ and C/EBPα protein expression. Confluent 3T3-L1 preadipocytes were differentiated into adipocytes in medium with or without different concentrations of resveratrol-amplified grape skin extracts (RAGE) for 8 days. Non, undifferentiated cells; DM, differentiated media cells. Values are mean ± SD (n = 3). Test for DM vs. Non: †††P < 0.001. Test for RAGE vs. DM: **P < 0.01, ***P < 0.001.

• 

  Fig. 6 Effect of resveratrol-amplified grape skin extracts on adipogenesis gene expression. Effects of the resveratrol-amplified grape skin extracts on adipogenic gene (SREBP1c, FAS, aP2, SCD-1, and LPL) expression during 3T3-L1 cell differentiation for 8 days. The results represent three independent experiments; actin was used as the internal control. Values are mean ± SD (n = 3). Test for DM vs. Non: ††P < 0.01, †††P < 0.001. Test for RAGE vs. DM: *P < 0.05, **P < 0.01, ***P < 0.001. DM, differentiated media cells; SREBP1c, adipocyte determination and differentiation dependent factor 1; aP2, adipocyte fatty acid binding protein; FAS, fatty acid synthetase; SCD-1, stearyl-CoA-desaturase-1; LPL, lipoprotein lipase.

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