Therapeutic Effect And Its Molecular Mechanism Of Natural Products D182in Mice Enteritis

Toll-like receptors are an important class of proteins involved in innate immunity, which also act as a bridge between innate and adaptive immunity. Ten TLRs have been identified in humans and12in mice, which are widely distributed in a variety of cell surface,such as monocytes macrophages, granulocytes, dendritic cells, lymphocytes, endothelial cells,epithelial cells and so on.Under certain conditions, TLRs abnormal expression or over-activation results in dysfunction of the body, causing a variety of diseases, such as inflammatory bowel disease (IBD), systemic lupus erythematosus (SLE), rheumatoid arthritis (AR). Therefore, searching new small molecule compound for blocking of TLRs signaling pathway can be expected for the treatment of TLRs-mediated immune diseases.D182, the symbiotes product of a marine origin, our previous study found D182had a therapeutic effect on LPS-induced mouse model of sepsis.In this article, we study its treatment of DSS-induced murine colitis and its molecular mechanism.1、D182could ameliorate DSS-induced mice acute colitis.Inflammatory bowel disease (IBD) is the local inflammation caused by many factors, and TLRs signaling pathway has a key role in the pathogenesis of IBD. In order to explore the anti-inflammatory effect in the body of the D182, we construct a DSS-induced acute colitis model D182on inflammatory bowel disease improvement. The results showed that D182can significantly reduce the weight loss in mice, a significant reduction in the level of blood in the stool, inhibit colon parts related inflammatory cytokine production and improve parts of the colon histopathological damage, showing a good therapeutic effect.2、D182could inhibit the expression of pro-inflammatory mediators.First, we used the CCK-8kit to detect the effect of D182in RAW264.7cells and mouse peritoneal macrophages cell viability, the results showed that D182in the10～100uM concentration range didn’t affect RAW264.7cells and mouse peritoneal macrophages cell viability. Second,we used Griess reagent detected the effect of D182in various of TLRs ligands-induced RAW264.7cells NO production, we found that D182inhibited a variety of TLRs ligands-induced RAW264.7NO production.Finally,we used Q-PCR and ELISA methods detected the effect of different concentrations of D182in PAM、LPS、PGN-induced RAW264.7cells and mouse peritoneal macrophages pro-inflammatory mediators mRNA and protein expression, Q-PCR datas showed D182dose-dependent inhibition of PAM、LPS、 PGN-induced the RAW264.7expression of TNF-alpha, IL-6. ELISA assay results showed that D182(5,15,45uM) in a dose-dependent inhibition of PAM、LPS、 PGN-induced RAW264.7cells and mouse peritoneal macrophages TNF-a production.3、The molecular mechanisms of small molecule compounds D182in treated enteritis.Inflammatory bowel disease (IBD) is the local inflammatory reponse caused by many factors.TLRs signaling pathway has a key role in the pathogenesis of IBD. In order to explore the anti-inflammatory effect of D182in vivo, we constructed DSS-induced acute colitis model and analyzed whether D182could improve inflammatory bowel disease. The results showed that the D182could significantly reduce the weight loss in mice, a significant reduction the level of blood in the stool, inhibition inflammatory cytokine production and improvement colon tissues of pathological damage and showed a good therapeutic effect.In summary, we have demonstrated for the first time that the small molecule compounds D182can effectively inhibit macrophage activation mediated by TLRs ligand, and the binding of D182with IRAK4active center maybe one of the reasons,we also certificated the anti-inflammatory effect of D182in DSS-induced mice acute colitis, All of these shows D182is a promising drug candidate for the treatment of inflammatory diseases.