Anti-adipogenic Effect of delphinidin-3-O-β-glucoside in 3T3-L1 Preadipocytes and Primary White Adipocytes (P06-018-19)

Abstract

ObjectivesDelphinidin-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. MethodsThe 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. ResultsTreatment 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. ConclusionsOur 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 SourcesThis work was supported by “Cooperative Research Program of Center for Companion Animal Research (Project No. PJ01398402)” Rural Development Administration, Republic of Korea.