Effects Of Panax Notoginseng Saponins On Calcium Balance And Growth Of Hepatocellular Carcinoma Cell Line HepG2

ObjectThe human hepatocarcinoma cell line HepG2 was treated by different concentrations of Panax notoginseng saponins at different time points to detect the growth inhibition rate,apoptosis rate and intracellular calcium concentration of human hepatoma cells.The possible mechanism provides a theoretical basis for the development of anti-hepatocarcinoma drugs.MethodThe human hepatoma cell line HepG2 cells were treated with 50 ug/ml,100 ug/ml,200 ug/ml,400 ug/ml and 800 ug/ml of the total saponin solution of Panax notoginseng,and HepG2 cells dealed with solvent was used as a control group.The cells in each group were cultured for 24 h,48h and 72 h.MTT assay was used to detect the inhibition rate of total saponins of Panax notoginseng on human hepatoma cell line HepG2 at different time points.Flow cytometry was performed to detect the effect of Panax notoginseng saponins on the apoptosis rate of human hepatoma cell line HepG2 by Annexin V and PI double staining method.After staining with Fura-2 AM calcium ion fluorescent probe,the intracellular calcium ion concentration of human hepatoma cell line HepG2 was determined by laser confocal microscopy.All data were statistically analyzed using SPSS 19.0.After one-way anova test,P < 0.05 was used as a significant difference.Result(1)Panax notoginseng saponins could inhibit the proliferation of human hepatoma cell line HepG2 cells.The human hepatoma cell line HepG2 cells were detected by MTT assay after dealing with different drug concentrations(50,100,200,400,800 ug/L).Compared with the control group,the proliferation inhibition was gradually observed,which was statistically different from the control group(P<0.05).At the same time,the effect time was from 24 to 72 hours.The total saponins of Panax notoginseng with the same concentration and different time were compared and finally we found that with the prolongation of time,the inhibition of human hepatoma cell line HepG2 cells were also significantly different(P<0.05),and showed a dose-effect and time-effect relationship.(2)Panax notoginseng saponins can induce apoptosis of human hepatoma cell line HepG2.The results of flow cytometry Annexin V/PI double staining showed that with the drug obtained by concentrations(50,100,200,400,800 ug/L)and treated with total saponins of Panax notoginseng from 24 h to 48 h,the apoptosis rate of human hepatoma cell line HepG2 was higher than that of the control group.The higher the concentration of total saponins of Panax notoginseng,the higher the apoptotic rate.The difference in the control group was statistically significant(P<0.05),and the apoptotic rate was more obvious when the total saponin concentration of Panax notoginseng was above 200ug/ml.(3)After treatment with the total saponins of Panax notoginseng,the concentration of intracellular calcium in the human hepatoma cell line HepG2 cells increased.When the concentration of total saponins of Panax notoginseng reached 400ug/ml and above,the fluorescence intensity was more obvious,indicating that the intracellular calcium concentration enhanced with the increase of PNS concentration.Conclusion(1)Panax notoginseng saponins can inhibit the growth of human hepatoma cell line HepG2 cells and induce apoptosis,and it is concentration-and time-dependent.The mechanism may be to inhibit tumors by affecting the intracellular calcium concentration of HepG2 cells.(2)The anti-hepatocarcinoma effect of Panax notoginseng saponins in vitro may provide a theoretical basis for animal and clinical trials.Panax notoginseng saponins could significantly increase anti-liver cancer effect through synergetic effect.