Figure 4

Rg3 diminishes TG accumulation in mature adipocytes with enhancement of fatty acid oxidation and mitochondrial maximal respiration. 3T3-L1 cells were seeded on the second day before differentiation and induced to become mature adipocytes. Fully differentiated adipocytes were treated with different concentrations of Rg3 (0–60 μM) for 7 d; (A) ORO quantification and representative images of ORO (magnification, 4×); (B) RT-PCR of the relative gene expression levels of Pparγ, Ap2, Fas, and Scd1; (C) Fatty acid (FA) oxidation rate (conversion of [³H]-oleic acid into [³H]-H₂O) in mature adipocytes incubated with 60 μM of Rg3 for 48 h; (D) FA oxidation rate in HepG2 cells. HepG2 cells were incubated with 60 μM of Rg3 for 48 h before exposure to 0.8 mM of BSA-oleic acid (OA) complex for 2 h; (E) Oxygen consumption rate (OCR) was determined by a Seahorse extracellular analyzer. The respiratory inhibitors, Oligo, FCCP, and a combination of Rot/AA are indicated with arrows; (F) OCR profiles; 3T3-L1 mature adipocytes were treated with 60 μM of Rg3 for 3–4 d. Cells were starved in DMEM for 12-18 h, followed by LPS (100 ng/ml) stimulation in 3T3-L1 adipocytes with or without Rg3. (G) RT-PCR of the relative gene expression levels of Il-6, Il-1β, and Tnfα; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 compared to the vehicle (Veh) control using Student’s t-test or one-way ANOVA with Bonferroni’s comparison test.