The Regulation Of TLR4 Pathway In Oxidation-damaged HUVE-12 Induced Activation And Migration Of M?

Part ? The effect of proliferation,migration and foam cell formation of M? induced by LPC-injured HUVE-12Objective:To observe whether Human umbilical vein endothelial cells(HUVE-12)injured by lyso-Phosphatidylcholine(LPC)can affect the proliferation,migration and foam cell formation of Macrophages(M?)derived from human acute monocytic leukemia cell line(THP-1).Methods:1?The oxidation-damaged HUVE-12 was prepared by LPC,and 0.4?m Transwell was used for co-culture of oxidation-damaged endotheial cell and THP-1-derived Macrophages(M?).2?Proliferation,migrationand foam cell formation of M? were detected by MTT assay,scratch assay,Oil red O staining and total cholesterol assay.The expression level of NF-kBp65,Myd88?TLR4 was detected in HUVE-12 cells by western blot.Results:LPC increased generation of reactive oxygen species(ROS)and leakage of lactate dehydrogenase(LDH)in HUVE-12 cells in concentration-dependent manner;LPC injured HUVE-12 increased proliferation,migration and foam cell formation of THP-1-derived M?.Part ? The effect of DFMG on proliferation,migration and foam cell formation of M? induced by oxidation-damaged HUVE-12Objective:To investigate the effect of 7-Difluoromethyl-5,4'-dimethoxygenistein(DFMG)on proliferation,migration and foam cell formation of THP-1-derived M? induced by oxidation-damaged HUVE-12.Methods:oxidation-damaged HUVE-12 cells/M? co-culture model was prepared which intervened by concentration gradient DFMG.Proliferation,migration and foam cell formation of THP-1-derived M? were detected by MTT assay,scratch assay,Oil red O staining and total cholesterol assay.The expression level of NF-kBp65,Myd88m?TLR4 was detected in HUVE-12 cells by western blot.Results:DFMG reduced proliferation,migration and foam cell formation of M? which was increased by oxidation-damaged HUVE-12 in concentration-dependent.DFMG down-regulated expression of TLR4?Myd88?NF-kBp65 in HUVE-12,and reduced concentration of IL-1? in co-culture condition medium.Part ? The effects of DFMG on proliferation,migration and foam cell formation of M? induced by oxidation-damaged HUVE-12 involved in TLR4 pathwayObjective:To explore wether inhibitory effects of DFMG on proliferation,migration and foam cell formation of THP-1 derived M? induced by oxidation-damaged HUVE-12 cells are involved in the modification of Toll like 4 receptor(TLR4)pathway.Methods:oxidation-damaged HUVE-12/M? co-culture model was prepared which intervened by CLI-095,Lipopolysaccharides(LPS,Interleukin-1 receptor antagonist(IL-1Ra),DFMG.Proliferation,migrationand foam cell formation of THP-1-derived M? were detected by MTT assay,scratch assay,Oil red O staining and total cholesterol assay.The expression level of NF-RBp65,Myd88?TLR4 was detected by western blot.Results:CLI-095 and IL-1Ra has the similar to effect of DFMG,reduced proli feration,migration and foam cell formation of M? which was increased by ox idation-damaged HUVE-12.LPS promoted LPC injured HUVE-12 induced proli feration,migration and foam cell formation of THP-1-derived of MO,and down-regulated expression of TLR4?Myd88?NF-kBp65 in HUVE-12,and reduced concentration of IL-1? in co-culture condition medium.LPS did the opposite.Conclusions:1.Proliferation,?migration and foam cell formation of M? by LPC-induced injured HUVE-12 cells.2.DFMG inhibited proliferation,migration and foam cell formation of M? by LPC-induced injured HUVE-12 cells.3.DFMG inhibited proliferation,migration and foam cell formation of THP-1-derived M? by LPC-induced injured HUVE-12 cells is involved in blockage of TLR4 signaling.