灯盏花甲素对脂多糖致小鼠RAW264.7巨噬细胞炎症反应的抑制作用及机制OA

Aim To study the inhibitory effect of api-genin-7-O-glucuronide(AG)on lipopolysaccharide(L PS)-induced inflammatory response of macrophage RA W264.7 and its mechanism.Methods RAW264.7 cells were pretreated with different concentrations of AG(10,5,2.5 μmol·L-1)for 2 h,and then exposed to LPS for different time points.The nucleus transcrip-tional activity of NF-κB was measured by dual lucifer-ase reporter assay.The fluorescence intensity of nucleus NF-κB p65 was detected by immunofloures-cent technique.The mRNA levels of TLR4,p38,ERK,JNK,NF-κB p65,IKK,IκB,STAT3,TNF-α,and IL-6 were detected by qPCR.The changes of ROS contents in cells were detected by high-content technology.The contents of inflammatory mediators TNF-α,IL-6,IL-1β were detected by ELISA.The NO content was detected using Griess method.TLR4,p38,ERK,JNK,IKK,IκB,p65,STAT3 protein and their phosphorylation were detected using Western blot.Results Compared with the model group,AG inhibited the phosphorylation of p38,ERK,JNK,IKK,IκB,p65,and STAT3,downregulated the mRNA expression of P38,ERK,JNK,IKK,IκB,P65,STAT3,TNF-α,IL-6,and TLR4,reduced nuclear translocation of NF-κB p65 and its transcrip-tional activity,decreased ROS levels,suppressed the secretion of inflammatory mediators(TNF-α,IL-6,NO,IL-1β),and downregulated the expression of TLR4 proteins.Conclusions AG can inhibit the in-flammatory response of macrophages induced by LPS.The underlying mechanism is related to the reduction of ROS release and the inhibition of TLR4/MAPK/NF-κB and STAT3 signaling pathway.