调脾护心方通过AKT/mTOR通路促进自噬抑制巨噬细胞泡沫化及焦亡的作用研究OA

Objective:To investigate the effect and mechanism of Tiaopi Huxin Decoction on oxidized low-density lipoprotein(ox-LDL)-induced macrophages foaming and pyroptosis.Methods:The human mononuclear-macrophage cell line THP-1 was cultured in vitro.It was stimulated and induced to become foam cells by ox-LDL.The appropriate administration concentration of the Tiaopi Huxin Decoction for macrophages was determined by the MTT colorimetric method.Different concentrations of the Tiaopi Huxin Decoction were used for intervention.Intracellular lipid deposition was observed by oil red O staining.Intracellular cholesterol levels were determined by enzymatic method.The protein expressions of ABCA1,ABCG1,NLRP3,GSDMD,IL-1β,LC3,P62,p-AKT and p-mTOR in macrophages were detected by Western Blot.Autophagy flow was detected by double-labeled adenovirus.Autophagy was inhibited by the autophagy inhibitor 3-MA to observe whether the Tiaopi Huxin Decoction inhibited the foaming and pyroptosis of macrophages induced by ox-LDL by regulating autophagy.Results:Tiaopi Huxin Decoction(10 to 100 mg/L)was non-toxic to macrophages.Compared with the model group,Tiaopi Huxin Decoction could reduce lipid deposition and the contents of TC and CE in macrophages,up-regulate the protein ex-pressions of ABCA1 and ABCG1,and down-regulate the protein expressions of NLRP3,Cleaved GSDMD,Mature IL-1β,P62,p-AKT,and p-mTOR.The ratio of LC3 Ⅱ/LC3 Ⅰ was increased,and autophagic flux was significantly enhanced(P<0.05 or P<0.01).The effect of the 3-MA could reverse the effect of Tiaopi Huxin Decoction(P<0.05 or P<0.01).Conclusion:Tiaopi Huxin Decoction may enhance the autophagic activity of macrophages by inhibiting the AKT/mTOR signaling pathway,thereby promoting cholesterol effervesia to reduce the formation of foam cells and inhibiting ox-LDL-induced pyroptosis of macrophages.