Topical Area: Dietary Bioactive Components
Objectives : Delphinidin-3-O-β-glucoside (D3G) is the main anthocyanin with well-recorded health properties, present in black soybean, bilberries, fruits, and flowers. In this study, we investigated the effect of D3G on the differentiation in 3T3-L1cell line, white adipose tissue from mice and its underlying molecular mechanism.
The 3T3-L1 cells and Primary white adipocytes (PWATs, obtained from the subcutaneous fat tissue of 5- to 6-week-old male C57BL/6 mice) were treated with D3G (25, 50 and 100 μg/mL) from day 0 to day 7. After the adipocytes differentiation, cells were stained with Oil Red O to check accumulated lipids. The total RNA and protein in 3T3-L1 cells and PWATs were isolated, and quantification of gene expression and immunoblotting were performed.
Results : Treatment with D3G at 25, 50 and 100 µM/ml concentrations significantly inhibited the lipid droplets accumulation in a dose-dependent manner. Real-time PCR and western blotting analysis revealed the inhibition of adipogenesis by downregulating the expression of peroxisome proliferator-activated receptor gamma (PPARγ), sterol regulatory element-binding transcription factor 1 (SREBP1), CCAAT/enhancer-binding protein alpha (C/EBPα), while the expression of key lipogenic factor namely fatty acid synthase (FAS) was downregulated at the translational level. Moreover, the relative protein expressions of silent mating type information regulation 2 homolog 1 (SIRT1) and carnitine palmitoyltransferase-1 (CPT-1) were upregulated. In addition, D3G effectively augmented the activation of AMP-activated protein kinase (AMPK) and Acetyl-CoA carboxylase (ACC) suggesting the possible anti-adipogenic effect of D3G through the AMPK pathway in both 3T3-L1cell line and primary white adipocytes. Furthermore, the dorsomorphin treatment decreased the relative expression of the AMPK and the activator 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) increased the AMPK expression, while together with D3G the expression was augmented, suggesting its plausible role in the AMPK pathway.
Conclusions : Our data suggest that D3G supplementation attenuated the differentiation in 3T3-L1cell line and primary white adipocytes (PWATs) by activation of the AMPK pathway. The results suggest the potential therapeutic role of D3G for obesity management and treatment.
Funding Sources :
This work was supported by “Cooperative Research Program of Center for Companion Animal Research (Project No. PJ01398402)” Rural Development Administration, Republic of Korea.