Study On Britannin-induced Apoptosis Via Regulating ROS Levels Of Human Hepatocellular Carcinoma HepG2 Cells

Objectives:The main objectives of this study were as follows:1.to observe the effect of britannin on the proliferation of human hepatoma HepG2 cell line;2 to observe the effect of britannin on the apoptosis of human hepatoma HepG2 cell line;3.to explore the possible mechanism of the induction of apoptosis in human hepatoma HepG2 cell line by britannin.Methods:1.MTT method was used to observe the effect of britannin on the proliferation of human hepatoma HepG2 cell line.2.Using Hoechst 33342 staining,to observe the effect of britannin on human liver cancer HepG2 cell line in morphology;3.DCFH-DA probe method was used to observe the changes of ROS level in human hepatoma cell line.To determine the content of malondialdehyde(MDA),reduced glutathione(GSH)and the activity of superoxide dismutase(SOD)to evaluate the oxidative stress state of cells;4.Detecing the effect of britannin on the expression of specific proteins in human hepatoma HepG2 cell line.Results:1.The administration of britannin with different concentrations inhibits the proliferation in human hepatoma HepG2 cell line.2.The results of Hoechst 33342 staining showed that the apoptotic cells increased significantly after the treatment of britannin in human hepatocellular carcinoma HepG2 cell line.3.After the treatment of britannin in human hepatocellular carcinoma HepG2 cell line for 24 hours,the concentration of reactive oxygen species(ROS)increased,The intracellular malondialdehyde(MDA)content increased and the superoxide dismutase(SOD)activity declined;4.After the treatment with britannin in human hepatoma HepG2 cell line,the expression of bcl-2 protein decreased significantly,and the expression of bax increased significantly.After the treatment with N-acetyl-L-cysteine(NAC),the expression of bcl-2 protein increased,and the bax expression decreased.Conclusion:1 Britannin has the effect of inhibiting the proliferation of HepG2 cell line in vitro;2.Britannin promotes the apoptosis of human liver cancer HepG2 cell line.3.Britannin induces HepG2 Oxidative stress state,up-regulates the ROS levels to induce apoptosis in human liver cancer HepG2 cell line.