Anti-adipogenic Effects and Mechanisms of Ginsenoside Rg3 in Pre-adipocytes and Obese Mice

FIGURE 1. Ginsenoside Rg3 dose-dependently inhibits cell differentiation in 3T3-L1 cells. The cells stained with Oil-red O were dissolved in isopropanol and relative fat accumulation was measured by absorbance using Synergy H1 hybrid reader at 490 nm. Oil-Red O representative images of lipid accumulation and the average bar graph were shown (A, 40× magnification). Cell toxicity study was measure by a MTT assay (B). Changes by treatment were expressed as the percentage of dimethyl sulfoxide (DMSO) control. Data are expressed as mean ± SE, n = 4. ∗P < 0.05, ∗∗P < 0.01 vs. DMSO control.

FIGURE 2. Ginsenoside Rg3 dose-dependently suppresses protein expressions of PPAR-γ (A) and C/EBP-α (B) in 3T3-L1 cells. On day 10, cells treated with various concentrations of Rg3 were collected to measure PPAR-γ and C/EBP-α protein expressions by western blotting and normalized by β-actin expression. The same membrane was stripped using a stripping buffer and blocked and probed with a new primary antibody to detect the next specific protein. Changes by treatment were expressed as the percentage of DMSO control. Data are expressed as mean ± SE, n = 3. A set of representative images and bar graph were shown. ∗P < 0.05, ∗∗P < 0.01 vs. DMSO control.

FIGURE 3. Ginsenoside Rg3 reduces PPAR-γ mRNA expression in 3T3-L1 cells. On day 10, cells treated with various concentrations of Rg3 were collected to measure PPAR-γ mRNA expression by quantitative real time PCR and normalized by β-actin expression. The same membrane was stripped using a stripping buffer and blocked and probed with a new primary antibody to detect the next specific protein. Changes by treatment were expressed as the percentage of DMSO control. Data are expressed as mean ± SE, n = 3. ∗P < 0.05, ∗∗P < 0.01 vs. DMSO control.

FIGURE 4. Ginsenoside Rg3 dose-dependently attenuates protein expressions of fat packing proteins FAS (A) and perilipin (B) in 3T3-L1 cells. On day 10, cells treated with various concentrations of Rg3 were collected to measure FAS and perilipin protein expressions by western blotting and normalized by β-actin expression. The same membrane was stripped using a stripping buffer and blocked and probed with a new primary antibody to detect the next specific protein. Changes by treatments were expressed as the percentage of DMSO control. Data are expressed as mean ± SE, n = 3. A set of representative images and bar graph were shown. ∗P < 0.05, ∗∗P < 0.01 vs. DMSO control.

FIGURE 5. Ginsenoside Rg3 dose-dependently reverses lipid accumulation, protein expressions of differentiation markers in HPAs. The cells stained with Oil-red O on day 15 were dissolved in isopropanol and fat accumulation was measured by absorbance using Synergy H1 hybrid reader at 490 nm. Oil-Red O representative images of lipid accumulation and the average bar graph were shown (A, 200× magnification); Protein expressions of PPAR-γ (B), C/EBP-α (C), FABP4 (D), and FAS (E) were determined by western blotting and normalized to β-actin expression. The same membrane was stripped using a stripping buffer and blocked and probed with a new primary antibody to detect the next specific protein. Changes by treatment were expressed as the percentage of DMSO control. Data are expressed as mean ± SE, n = 3. A set of representative images and bar graph were shown. ∗P < 0.05, ∗∗P < 0.01 vs. DMSO control.

FIGURE 6. In vivo effects of dietary ginsenoside Rg3 intake. After 8-week dietary administration of Rg3 (0.1 mg/kg diet), the liver PPAR-γ (A) and C/EBP-α (B) protein expression in mice was determined by western blotting and normalized to β-actin expression. The same membrane was stripped using a stripping buffer and blocked and probed with a new primary antibody to detect the next specific protein. Body weight gain, food intake, fasting blood glucose, and hepatic antioxidant were measured and are shown in Table 1. Data are expressed as mean ± SE, n = 12. A set of representative images and bar graph were shown. ∗P < 0.05, ∗∗P < 0.01 vs. standard diet (SD) control.