Effects Of Simiao Yongan Decoction-containing Serum On OX-LDL-induced Autophagy And Inflammatory Response Of Foamy Macrophages

Objective:to investigate the effect of Si Miao Yong An decoction on OX-LDL induced autophagy and inflammatory response in vitro.Methods:1.SD rats were treated with Low,medium and high doses of Si Miao Yong An decoction by gavage.2.Cell proliferation toxicity assay(CCK8)was used to observe the effect of drug-containing serum on macrophage proliferation.3.Lipid droplet changes of macrophages were observed by oil red O staining.4.IL-6,IL-10 and IFN-y inflammatory cytokines were detected by ELISA.5.To observe the effect of Si Miao Yong An decoction on autophagy of macrophage induced foam cells in vitro.6.The expression of cell immune fluorescence labeling method to detect LC3-II.7.Expressions of p-AMPK?p-mTOR?p-p70S6K were detected by Western blot.Results:1.The results of CCK8 showed that the OD value of macrophages was not significantly changed in Si Miao Yong An decoction with the concentration of less than 20%.2.Oil red O staining showed that after OX-LDL modeling intervention,a large number of orange lipid droplets were seen in the cytoplasm of macrophages.3.ELISA showed that the serum containing Si Miao Yong An decoction reduced the levels of IL-6 and IFN-? inflammatory cytokines and promoted the secretion of IL-10(P<0.05).4.The results of immunofluorescence analysis showed that the expression of LC3-II in the serum treated group was increased.5.The observation under transmission electron microscopy showed that the increase of autophagosomes was observed in foam cells after the intervention of Si Miao Yong An decoction.6.Western blot results showed that the expression of p-mTOR and p-p70S6K was inhibited and the expression of p-AMPK was up-regulated in the frothing macrophages in each group treated with Si Miao Yong An decoction(P<0.05).Conclusion:Si Miao Yong An decoction can reduce the expression of inflammatory factors in froth cells,and possibly regulate the AMPK/mTOR/p70S6K signaling pathway to up-regulate the expression of autophagy related gene LC3-II,induce autophagy in froth macrophages,and thus play a role in the treatment of AS.