The Intracellular Signaling Mechanism Of HMGB1amplying The Inflammation Induced By LPS In Macrophage

Objective:To study the mechanism of HMGB1amplified LPS-induced inflammatory response,。Method:RAW264.7cells were treated with LPS (100ng/ml)/LPS+HMGB1(100ng/ml) for0、2hours,Immunofluorescence was used to measure the expression of NF-KB;RAW264.7cells were treated with LPS (100ng/ml)/LPS+HMGB1(100ng/ml)for0、15、30、60minutes to measure the level of ikb-a, p-p38and p-IRF3by western blot; RAW264.7cells were treated with LPS(100ng/ml) or LPS+HMGB1(100ng/ml) for0、3、6hours, real time PCR (RT-PCR) was used to detect the expression of IL-6and IFN-βmRNA; RAW264.7cells were treated with10uM P38MAPK inhibitor in advance, RAW264.7cells were processed by the method of Appeal,we use RT-PCR to detect the expression of IL-6and IFN-β mRNA., and western-blot to detect the level of phosphorylation of p-IRF3.Result:1.HMGB1amplified the level of IL-6and IFN-β mRNA in LPS-induced RAW264.7Macrophage(P<0.05);2.HMGB1had no significant effect on ikb-a degradation and NF-KB activation in LPS-induced Macrophages (p>0.05);3.HMGB1up-regulated the level of P-IRF3and P-P38in LPS-induced Macrophages (P<0.05);4. P38MAPK inhibitor down-regulated the expression of IL-6and IFN-β mRNA in LPS or LPS+HMGB1stimulated Macrophages (P<0.05);. P38MAPK inhibitor also down-regulated the level of phosphorylation of p-IRF3in LPS+HMGB1co-stimulated Macrophages(P<0.05).Conclusion:HMGB1could amplify LPS-induced TLR4signaling through P38MAPK/AP1and P38MAPK/IRF3pathway...