Figure 7.

An accumulation of malonyl-CoA and inhibition of CPT-1 activity participated in [6]-gingerol-induced apoptosis and inhibition of FASN in HepG2 cells. Cells were treated with 0.5 mM [6]-gingerol or 0.1 mM C75 or 10 µg/ml TOFA for 6 h. 10 µg/ml TOFA was pre-incubated for 1 h prior to [6]-gingerol or C75 treatment for 6 h. The control was defined as cells treated with a medium or 0.1%DMSO vehicle alone. A. Percentages of intracellular fatty acid levels were determined using the free fatty acid quantification kit. B. ΔΨm was measured by JC-1 dye fluorochrome and detected by flow cytometry. The disruption of ΔΨm was quantified by calculating an increase of a monomer JC-1 green fluorescence in the cytoplasm and a decrease of an aggregated JC-1 red fluorescence in the mitochondria. CCCP was used as a positive control to induce depolarization of ΔΨm. C. Cells were treated with 0.5 mM [6]-gingerol, 0.1 mM C75, and 10 µg/ml TOFA for 24 h. CPT-1 activity was assayed and presented as percentage of CPT-1 (nmol/min/mg protein). Three independent experiments were performed for statistical analysis and expressed as mean ± SD. *denotes statistically significant difference from the control at P<0.05.