Inhibitory Effects of Chung Hun Wha Dam Tang (CHWDT) on High-Fat Diet-Induced Obesity via AMP-Activated Protein Kinase Activation

Figure 6

CHWDT increases CPT1 expression and reduces lipid content through AMPKα activation in HepG2 cells.Cells were incubated with 100 𝜇g/mL CHWDT for 1, 12, and 24 h and 2 mM metformin (a) Cells were incubated with 50 and 100 𝜇g/mL CHWDT for 9 h and 2 mM metformin (b) Cells were pretreated with CHWDT for 1 h and palmitate (0.45 mM) was treated for 12 h and 2 mM metformin (c) Cells were treated with metformin for 3 h as positive control of AMPKα activation. Western blotting was performed to detect phospho-AMPK𝛼, total AMPKα and beta-actin as loading control. The representative blot is shown. Cells were pretreated with or without compound C (20 𝜇M) for 30 min and treated with CHWDT (50 and 100 𝜇g/mL) for 12 h. CPT1 mRNA was checked by RT-PCR, and representative band is shown (d) Intracellular triglycerides (TG) levels were detected by TG-S kit, where cells were treated with CHWDT (50 and 100 𝜇g/mL) in presence of palmitate (0.45 mM) for 24 h with or without preincubating with 20 𝜇M compound C for 30 min. The control cells were incubated with 1.76% bovine serum albumin (BSA) for 24 h (e) Data indicating mean ± SEM of three independent experiments with triplicate wells, 𝑃<0.01 as **, and 𝑃<0.001 as ***.

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