Screening Of Anti-inflammatory Components In Stellera Chamaejasme And Investigation Of Anti-inflammatory Therapeutic Effect Of The Components On Endotoxemia Animal Model

The present study screened anti-inflammatory components from35kinds ofcomponents purified from stellera chamaejasme via lipopolysaccharide (LPS)-inducedmouse peritoneal macrophages in vitro model of inflammation. Innate immunity is the firstbarrier of body against invading pathogens. Pattern recognition receptors（PRRs） such astoll-like receptors (TLRs) acts as primary sensors that detect a widely varity of microbialcomponents in the innate immune system. TLR4expressed on macrophages recogonize thelipopolysaccharide from Gram-negative bacteria and LPS is ligand of TLR4. Then TLR4together with MD-2, CD-14and LPS forms a complex, which activate intracellular signalingpathways that lead to the activation of MAP kinase (MAPK) and the nuclear transcriptionfactor (NF-κB) to induce the expression of inflammatory cytokines and mediators such asTNF-α,IL-6,IL-1β,iNOS and so on. All these different pathways are integrated into themacrophage response towards an inflammatory stimulus by a highly complex cross-talk ofthe pathways engaged. This process is tightly regulated by several intra-and inter-cellularfeedback loops to warrant an inflammatory response sufficient to battle invading pathogensand to avoid non-essential tissue damage caused by an overwhelming inflammatoryresponse.Anti-inflammatory properties of the diterpenoid components separated fromchamaejasme was investigated by LPS-induced mouse peritoneal macrophages in vitromodel of inflammation. TNF-αfrom the cell supernatant was evaluated by ELISA. Severalanti-inflammatory components were identified from30kinds of diterpenoid components instellera chamaejasme. It was shown that WYB-19significantly reduced the production ofpro-inflammatory cytokines such as tumor necrosis factor alpha (TNF-α). The mRNA levelof several inflammatory mediators such as TNF-α, IL-1β,iNOS and IL-6was tested by realtime PCR, the results indicated that WYB-19significantly inhibited the production ofpro-inflammatory cytokines and inflammatory mediators, such as TNF-α,IL-6,IL-1β,iNOS.Furthermore,intracellular signaling pathways were analyzed by Western blot usingspecific antibodies.The results suggested that WYB-19exerts an anti-inflammatory via inhibiting phosohorylation of p38MAPK and activition of NF-κB. Based on above resultsfrom the in vitro studies, we further investigated the therapeutic effect of WYB-19onendotoxemia mouse model. The results indicated that WYB-19significantly decerased themortility of endoxemia mouse in a dose-dependent manner. In addition, WYB-19is a lowtoxicity and high potency anti-inflammatory compounds with potential anti-inflammatorytreatment effect, which has important significance. The project has a important significanceon the development of novel drugs target inflammatory diseases such as endotoxemia andalso implicated on further development of chamaejasme.