Study The Effect And Mechanism Of Sanguinarine On Human Umbilical Vein Endothelial Cells

Objective:To investigate the inhibitory effect and mechanism of sanguinarine on HUVEC,and to provide a theoretical and experimental basis for the use of sanguinarine for anti-angiogenesisMethods:Human Umbilical Vein Endothelial Cells(HUVEC)were cultured and divided into control group and sanguinarine group.The control group was treated with physiological saline and the sanguinarine group was treated with 1.00?mol/L,2.00 ?mol/L,3.00 ?mol/L and 4.00 ?mol/L sanguinarine.After 48h,the effects of sanguinarine on proliferation and tubular formation of HUVEC cells were detected by MTT assay and tubular formation assay.Flow cytometry and Western blot were used to detect the cell cycle,the expressiont of apoptosis-related proteins Bax,Caspase-3,and Thromboinsensitive protein 1(TSP-1)in 1.00?mol/L and 2.00?mol/L sanguine concentrations.Results:1.Sanguinarine can significantly inhibit the proliferation of HUVEC in a dose-dependent manner.Compared with the control group,HUVEC were significantly inhibited by the concentration of blood alkali in each group,and the difference was statistically significant(P<0.05).With the increase of blood concentration,the inhibition rate increased,and the inhibition rate was positively correlated with the concentration.The correlation coefficient was R=0.067(P<0.05).Calculated the IC50 value of sanguinarine was 2.0366 ?mol/L.2.Sanguinarine can significantly inhibit the proliferation of HUVEC in a time-dependent manner.With the prolongation of the action time of sanguinarine,the inhibitory effect of cell proliferation was enhanced,and the inhibitory effect reached its peak at 48h.Compared with 72h and 48h,there was no significant difference in cell proliferation inhibition(P>0.05).3.Sanguinarine can promote the apoptosis of HUVEC.Compared with the control group,the concentration of 1.00?mol/L and 2.00?mol/L sodium bicarbonate can significantly increase the apoptosis rate of HUVEC,and the difference is statistically significant(P<0.05).<0.05).4.Sanguinarine can significantly affect the cell cycle of HUVEC.Compared with the control group,the percentage of G1 phase cells in the 2.0 ?g/ml sanguinarine group decreased,and the percentage of S phase cells increased,the difference was statistically significant(P<0.05).The percentage of each cycle in the 1.0 ?g/ml concentration group was not significantly different from that in the control group(P>0.05).5.Effect of sanguinarine on the expression of Bax,Capase-3 and TSP-1 proteins in HUVEC.Compared with the control group,the expression of Caspase-3,TSP-1 was significantly increased in sanguinarine 1.0?g/ml,2.0?g/ml group.The expression of Bax was significantly increased in the 2.0?g/ml group,and the difference was statistically significant(P<0.05).Conclusion:Sanguinarine can up-regulate the expression of Bax,Caspase-3 and TSP-1,inhibit the proliferation of HUVEC,promote the apoptosis of HUVEC,inhibit the formation of new blood vessels,and play a role in anti-angiogenesis.