Regulation Of Tiaozhi Granules On Autophagy Of HUVECs

Purpose:To investigate the role of Tiaozhi Granules in the autophagy of HUVECs on the cellular,protein and molecular level.Methods:HUVECs were incubated by Tiaozhi granule medicated serum,6-amino-3-methyl purine(3-MA)and rapamycin.Using MTT detected the cell viability with Tiaozhi granule medicated serum.Using SYBR Green I realtime PCR detected the expressive levels of mTOR?ATG5?ATG7?Beclin-lUsing western blot detected the expressive levels of LC3-? and Beclin-1.Immunofluorescence was used to check up changes in the numbers of autophagosomes.Results:The results showed that Tiaozhi Granules did not affect on the cell survival.The expression of each gene was up-regulated.Tiaozhi Granules increased the ratio of LC3-? to LC3-I(p<0.05).The number of LC3 autophagic bodies increased.But there was no significant difference in Beclinl expression.Tiaozhi Granules with rapamycin decreased the expression of each gene and the ratio of LC3-II to LC3-I(p<0.05 or p<0.01).The number of LC3 autophagic bodies decreased.There was no significant difference in Beclinl expression.3MA treatment group and rapamycin alone in the group are significantly different in different levels(p<0.05),but with the concentration of serum lipid particles combined with rapamycin treatment group had no significant difference(P>0.05).Conclusion:The results of this study showed that Tiaozhi Granules could regulate the autophagy of endothelial cells at protein level and molecule level.This may be one of the mechanisms that Tiaozhi Granules anti atherosclerosis and regulate blood lipids.There are differences in expression of various Tiaozhi granule drug group and control group related to LC3,but there was no significant difference in the expression of Beclinl,indicating that lipid particles through regulation and synthesis of LC3 related signal pathway to regulate autophagy in endothelial cells.