Artesunate induces apoptosis of human hepatocellular carcinoma HepG2 cells by increasing generation of reactive oxygen species / 中国病理生理杂志

ABSTRACT

AIM:To investigate the effect of reactive oxygen species(ROS)on the apoptosis of HepG2 cells induced by artesunate.METHODS:The effect of artesunate on the viability of HepG 2 cells was measured by MTT assay. The morphological changes of the apoptotic cells were observed by the method of Hoechst 33258 fluorescence staining.The apoptosis of HepG2 cells was analyzed by flow cytometry.DCFH-DA was used to detect the changes of ROS generation dur-ing the process of apoptosis.The protein levels of Bax ,Bcl-2,cleaved caspase-3 and cytochrome C(Cyt C)were deter-mined by Western blot.HepG2 cells were pretreated with apocynin and then Western blot was used to detect the expression of p47phox and p22phox,and ROS changes were analyzed by flow cytometry.RESULTS:Compare with control group,the cell viability was obviously inhibited after treatment with artesunate for 24 h(P<0.05).The nuclei were densely stained ,and the proportion of apoptotic cells was increased(P<0.05).ROS was increased significantly(P<0.05).The results of Western blot demonstrated that the expression level of Bax was increased ,Bcl-2 was decreased ,the ratio of Bax/Bcl-2 was increased ,and the protein levels of cleaved caspase-3 and Cyt C were increased.Pretreatment with apocynin reduced the expression of p47phox and p22phox and the generation of ROS in the artesunate treatment group.CONCLUSION:Artesu-nate induces the apoptosis of HepG 2 cells.The possible mechanism may be related to the increase in the generation of ROS.