Emodin induces apoptosis in human hepatocellular carcinoma HepaRG cells via the mitochondrial caspase-dependent pathway

Dong, Xiaoxv; Ni, Boran; Fu, Jing; Yin, Xingbin; You, Longtai; Leng, Xin; Liang, Xiao; Ni, Jian

plays an important role in the process of cell apoptosis. To measure the effect of emodin on the ROS level, the cells were stained with DCFH-DA and analyzed by flow cytometry. Compared with the control group, the results showed that the ROS level was increased dose-dependently in HepaRG cells after emodin treatment. However, after pre-treatment with NAC for 1 h, ROS generation was significantly attenuated (Fig. 4A and B) . Emodin reduced ΔΨm and induces cytochrome c release from mitochondria. To investigate the effect of emodin on mitochondrial functions of HepaRG cells. ΔΨm was measured using JC-1 fluorescence staining by flow cytometry. Treatment with emodin induced the loss of mitochondrial membrane potential (MMP ΔΨm) in HepaRG cells in a dose-dependent manner (Fig. 5A and B) . Furthermore, we investigated the release of cytochrome c by Western blot analysis. The level of cytochrome c was significantly increased in the cytosol fraction, while markedly decreased in the mitochondria after treatment with emodin for 24 h (Fig. 5C and D) . Taken together, emodin induces apoptosis through a change in MMP and regulation of apoptosis-related proteins in HepaRG cells.