| Inflammation is mediated inflammatory factors secreted by the immune system of a natural defense against harmful stimuli response;however the abnormal inflammatory reaction such as excessive or against their normal tissue reaction can cause tissue damage,leading to a variety of diseases,such as acute and chronic inflammatory diseases and autoimmune diseases.Therefore,control the inflammatory response is a great significance for the treatment of inflammation-related diseases.IL-6 is an important proinflammatory cytokines.This study is based on our previous work in which by using IL-6 genepromoter –drived luciferase reporter system,TI110 was identified to be able to effectively inhibit IL-6 expressionafter screening over 300 natural compounds.Here,TI110,as one of the anti-inflammation candidates was further studied.In order to study the inhibition of TI110 on inflammatory responses,we built the macrophage inflammatory model induced by LPS.Firstly,weconfirmed that TI110 at the dose used in this study did not affect the cell viability of macrophages with MTT assy.Then,by using RT-PCR,RT-qPCR and ELISA,TI110 was found to significantly reduce LPS-induced expression of the inflammatory cytokine IL-6,TNF-α and IL-1β at both RNA and protein levelsin RAW264.7 cells,mouse peritoneal macrophage and THP1 cells.Simutaneously,we found that TI110 couldpromote the mRNAexpression of anti-inflammatory cytokinessuch as IL-10 and TGF-β1 and increase their release in these macrophages.In addition,we studied the influence of TI110 on other proinflammatory factors and found that TI110 could obviously inhibit the expression of iNOS and NO release and suppress the expression of COX2 protein and HMGB1 secretion.Next we further investigated the mechanism by which TI110 elicits its anti-inflammatory effects on macrophages.TI110 could inhibit the NF-κB phosphorylation and p65 translocation into the nucleus in LPS induced macrophages,which suggested that TI110 could suppress LPS-stimulated activation of NF-κBpathway.In addition,TI110 significantly inhibited the expression of IRF5 in LPS induced macrophage as well.As for MAPK pathways,the p38/MAPK,ERK/MAPK signaling pathway were not affected while JNK/MAPK signaling pathway was inhibited by TI110.Furthermore,we also found that TI110 could activate AMPK pathway in LPS induced macrophages as shown as enhanced AMPK phosphorylation upon TI110 treatment.Inhibition of AMPK activation by its inhibitor compound C abolished TI110-induced inhibition on NF-κB and IRF5 indicating TI110 might inhibit NF-κB and IRF5 pathways by activating AMPK pathway.Together,these results suggested that TI110 exerted its anti-inflammtory effects through multiple mechanisms.Finally,we establisheda LPS-induced sepsis shock model in mice and found that intraperitoneal injection of TI110 could increase the survival rate of mice,reduce the production of inflammatory cytokines IL-6,TNF-α and IL-1β and release of NO and HMGB1 in serum,increase the secretion of anti-inflammatory factor IL-10 and alleviate the liver damage.In conclusion,we demonstrated that TI110 had significant anti-inflammatory effects in both in vitro and in vivo LPS induced inflammatory responses and it influenced multiple inflammation related signaling pathways.Thus,natural compound TI110 may be a potential drug for the treatment of inflammatory diseases. |
