Lupinol-3β-succinate induce apoptosis of HepG2 cells by regulating the mitochondrial apoptosis pathway
Objective To investigate the effect of lupinol-3β-succinate on apoptosis of HepG2 cells and mechanism.Methods HepG2 cells were treated with lupinol-3β-succinate(0,2.5,5,10,20,40 and 80 μmol/L)for 24,48,and 72 h,and the proliferation activity of the cells was detected by CCK-8 method.HepG2 cells were treated with lupinol-3β-succinate(0,4,8,and 16 μmol/L)for 48 h,and the apoptosis rate of each group was detected by AO/EB staining and Annexin V-FITC/PI double staining.Changes of mitochondrial membrane potential in HepG2 cells were observed by JC-1 staining.Levels of ROS were observed by DCFH-DA staining.Western blotting method was used to detect the changes of mitochondrial apoptosis-related protein expression in each group.Results Lupinol-3β-succinate showed significant time and dose dependent inhibition of HepG2 cell proliferation(P＜0.01).Compared with control group,ROS level,total apoptosis rate,Bax,Cyt-C,cleaved Caspase-9,and cleaved Caspase-3 protein levels were increased in a dose-dependent manner after lupinol-3β-succinate treatment(P＜0.01,0.001).Mitochondrial membrane potential and Bcl-2 protein expression were significantly decreased(P＜0.01).Conclusion Lupinol-3β-succinate may induce apoptosis of human hepatoma HepG2 cells by regulating mitochondrial apoptosis pathway,and finally play an anti-hepatocellular carcinoma role.
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