J Nutr Health.  2019 Feb;52(1):17-25. 10.4163/jnh.2019.52.1.17.

Effects of Lonicera caerulea extract on adipocyte differentiation and adipogenesis in 3T3-L1 cells and mouse adipose-derived stem cells (MADSCs)

• Park M
• Lee C
• Lee HJ

• KMID: 2466679
• DOI: http://doi.org/10.4163/jnh.2019.52.1.17

Abstract

PURPOSE
Obesity is a major health problem of global significance because it is clearly associated with an increased risk of health problems, such as nonalcoholic fatty liver disease (NAFLD), diabetes, cardiovascular diseases, and cancer. Lonicera caerulea (LC) originates from high mountains or wet areas and has been used as a traditional medicine in northern Russia, China, and Japan. LC contains a range of bioactive constituents, such as vitamins, minerals, and polyphenols. This study examined the anti-obesity effects of LC during differentiation in preadipocytes.
METHODS
The cell viability assay was performed after the differentiation of 3T3-L1 cells for 7 days. Oil Red O staining was used to visualize the changes in lipid droplets in 3T3-L1 cells and mouse adipose-derived stem cells (MADSCs). The mRNA expression of obesity-related genes was determined by quantitative real-time PCR.
RESULTS
According to the results of Oil Red O staining, the lipid levels and size of lipid droplets in the adipocytes were reduced and the LC extract (LCE, 0.25-1 mg/mL) markedly inhibited adipogenesis in a dose-dependent manner. The treatment of LCE also decreased the mRNA expression of peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein-α (C/EBPα), and sterol regulatory element binding protein 1 (SREBP1) in 3T3-L1 cells. Western blot analysis showed that the PPARγ, C/EBPα, and SREBP1 protein levels in both 3T3-L1 and MADSC were reduced in a dose-dependent manner.
CONCLUSION
These results suggest that LCE can inhibit adipogenic differentiation through the regulation of adipogenesis-related markers.

Keyword

Lonicera caerulea; preadipocyte; anti-obesity; lipid accumulation

MeSH Terms

3T3-L1 Cells*
Adipocytes*
Adipogenesis*
Animals
Blotting, Western
Cardiovascular Diseases
Cell Survival
China
Japan
Lipid Droplets
Lonicera*
Medicine, Traditional
Mice*
Minerals
Miners
Non-alcoholic Fatty Liver Disease
Obesity
Peroxisomes
Polyphenols
Real-Time Polymerase Chain Reaction
RNA, Messenger
Russia
Stem Cells*
Sterol Regulatory Element Binding Protein 1
Vitamins
Minerals
Polyphenols
RNA, Messenger
Sterol Regulatory Element Binding Protein 1
Vitamins

Figure

• 

  Fig. 1 Effect of Lonicera caerulea extract (LCE) on 3T3-L1 preadipocytes proliferation. 3T3-L1 preadipocyte cells were treated with the indicated concentrations of LCE for 24, 48, and 72hr and cell viability was estimated by a CCK-8 assay. All experiments were repeated at least three times and data represent means ± SD. ** p < 0.01, *** p < 0.001 vs. Con.

• 

  Fig. 2 Lipid accumulation was determined by Oil Red O staining. LCE treated 3T3-L1 adipocytes (A) and lipid levels in 3T3-L1 adipocytes (B). Original magnification 200X. Data represent means ± SD. ** p < 0.01, *** p < 0.001 vs. Con.

• 

  Fig. 3 Lipid accumulation was determined by Oil Red O staining. LCE treated mouse adipose-derived stem cell (MADSC) adipocytes (A) and lipid levels in MADSC adipocytes (B). Original magnification 200X. Data represent means ± SD. *** p < 0.001 vs. Con.

• 

  Fig. 4 Effects of LCE on the expression of genes associated with adipogenesis in 3T3-L1 cells. The expression of Pparγ (A), C/ebpα (B), and Srebp1 (C) were quantified by real-time PCR and normalized by β-actin as an internal control. (D) Western blot analysis of PPARγ, C/EBPα, and SREBP1 protein. Data represent means ± SD. * p < 0.05, ** p < 0.01 vs. Con.

• 

  Fig. 5 Effects of LCE on the expression of protein associated with adipogenesis in MADSCs. The protein expression of PPARγ (A), C/EBPα (B), and SREBP1 (C) were normalized by β-actin as an internal control. (D) PPARγ, C/EBPα, and SREBP1 protein levels by Immunoblot analysis. Data represent means ± SD. * p < 0.05, ** p < 0.01 vs. Con.

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