Nutr Res Pract.  2015 Aug;9(4):439-444. 10.4162/nrp.2015.9.4.439.

Soluble extract of soybean fermented with Aspergillus oryzae GB107 inhibits fat accumulation in cultured 3T3-L1 adipocytes

• So KH
• Suzuki Y
• Yonekura S
• Suzuki Y
• Lee CH
• Kim SW
• Katoh K
• Roh SG

• KMID: 2313863
• DOI: http://doi.org/10.4162/nrp.2015.9.4.439

Abstract

BACKGROUND/OBJECTIVES
This study was conducted to investigate the effects of fermented soybean (FS) extract on adipocyte differentiation and fat accumulation using cultured 3T3-L1 adipocytes.
MATERIALS/METHODS
3T3-L1 adipocytes were treated with FS and nonfermented soybean (NFS) extract during differentiation for 10 days in vitro. Oil red O staining was performed and glycerol-3-phosphate dehydrogenase (GPDH) activity was measured for analysis of fat accumulation. Expressions of adipogenic genes were measured.
RESULTS
Soluble extract of soybean fermented with Aspergillus oryzae GB107 contained higher levels of low-molecular-weight protein than conventional soybean protein did. FS extract (50 microg/ml) inhibited adipocyte differentiation and fat accumulation during differentiation of 3T3-L1 preadipocytes for 10 days in vitro. Significantly lower GPDH activity was observed in differentiated adipocytes treated with the FS extract than those treated with NFS extract. Treatment with FS extract resulted in decreased expression levels of leptin, adiponectin, and adipogenin genes, which are associated with adipogenesis.
CONCLUSIONS
This report is the first to demonstrate that the water-soluble extract from FS inhibits fat accumulation and lipid storage in 3T3-L1 adipocytes. Thus, the soybean extract fermented with A. oryzae GB107 could be used to control lipid accumulation in adipocytes.

Keyword

Fermented soybean; adipocyte; adipogenesis; 3T3-L1; Aspergillus oryzae

MeSH Terms

Adipocytes*
Adipogenesis
Adiponectin
Aspergillus oryzae*
Glycerolphosphate Dehydrogenase
Leptin
Oryza
Soybeans*
Adiponectin
Glycerolphosphate Dehydrogenase
Leptin

Figure

• 

  Fig. 1 Distribution of the water-soluble proteins extracted from FS and NFS. Approximately 25 µg of protein was loaded per lane. Lanes M contain Marker.

• 

  Fig. 2 Effect of (A) FS and (B) NFS on the viability of 3T3L-1 preadipocytes by MTT assay. Values are expressed as a percentage survival compared to control (0 µg/ml) after 24 h incubation.

• 

  Fig. 3 Morphological changes in 3T3-L1 cells cultured with NFS extract (50 µg/ml) or FS extract (50 µg/ml). (A) After 10 days, the cells were fixed and stained with oil red O for detection of oil droplets. (B) Magnified photographs of 3T3-L1 adipocytes cultured with NFS extract (50 µg/ml) or FS extract (50 µg/ml). (C) The stained cells were destained with isopropanol, and the OD of the de-staining isopropanol was measured by spectrophotometry. Values are expressed as mean ± SEM (n = 6). abMean values with different superscripts are significantly different (P < 0.05).

• 

  Fig. 4 GPDH activity of 3T3-L1 adipocytes cultured with NFS (50 µg/ml) or FS extract (50 µg/ml) for 10 days. Values are expressed as mean ± SEM (n = 6). abMean values with different superscripts are significantly different (P < 0.05).

• 

  Fig. 5 Glycerol release from 3T3-L1 adipocytes on days 0, 2, 4, 6, 8, and 10 after induction of adipocyte differentiation. 3T3-L1 cells were cultured with NFS extract (50 µg/ml) or FS extract (50 µg/ml) after induction of differentiation. Data were normalized with respect to 100% for the control at day 0. Values are expressed as mean ± SEM (n = 3). abMean values with different superscripts are significantly different (P < 0.05).

• 

  Fig. 6 Expression levels of adipogenic-associated genes in 3T3-L1 adipocytes cultured with NFS (50 µg/ml) or FS (50 µg/ml) extracts for 10 days. The data were normalized to β-actin mRNA levels, and values are expressed as the fold-value of that obtained for the control. Values are expressed as mean ± SEM (n = 3). abMean values with different superscripts are significantly different (P < 0.05).

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