RRC ID 35999
著者 Wang Y, Shan X, Chen G, Jiang L, Wang Z, Fang Q, Liu X, Wang J, Zhang Y, Wu W, Liang G.
タイトル MD-2 as the target of a novel small molecule, L6H21, in the attenuation of LPS-induced inflammatory response and sepsis.
ジャーナル Br J Pharmacol
Abstract BACKGROUND AND PURPOSE:Myeloid differentiation 2 (MD-2) recognizes LPS, which is required for TLR4 activation, and represents an attractive therapeutic target for severe inflammatory disorders. We previously found that a chalcone derivative, L6H21, could inhibit LPS-induced overexpression of TNF-α and IL-6 in macrophages. Here, we performed a series of biochemical experiments to investigate whether L6H21 specifically targets MD-2 and inhibits the interaction and signalling transduction of LPS-TLR4/MD-2.
EXPERIMENTAL APPROACH:The binding affinity of L6H21 to MD-2 protein was analysed using computer docking, surface plasmon resonance analysis, elisa, fluorescence measurements and flow cytometric analysis. The effects of L6H21 on MAPK and NF-κB signalling were determined using EMSA, fluorescence staining, Western blotting and immunoprecipitation. The anti-inflammatory effects of L6H21 were confirmed using elisa and RT-qPCR in vitro. The anti-inflammatory effects of L6H21 were also evaluated in septic C57BL/6 mice.
KEY RESULTS:Compound L6H21 inserted into the hydrophobic region of the MD-2 pocket, forming hydrogen bonds with Arg(90) and Tyr(102) in the MD-2 pocket. In vitro, L6H21 subsequently suppressed MAPK phosphorylation, NF-κB activation and cytokine expression in macrophages stimulated by LPS. In vivo, L6H21 pretreatment improved survival, prevented lung injury, decreased serum and hepatic cytokine levels in mice subjected to LPS. In addition, mice with MD-2 gene knockout were universally protected from the effects of LPS-induced septic shock.
CONCLUSIONS AND IMPLICATIONS:Overall, this work demonstrated that the new chalcone derivative, L6H21, is a potential candidate for the treatment of sepsis. More importantly, the data confirmed that MD-2 is an important therapeutic target for inflammatory disorders.
巻・号 172(17)
ページ 4391-405
公開日 2015-9-1
DOI 10.1111/bph.13221
PMID 26076332
PMC PMC4556476
MeSH Animals Cell Line Chalcone / administration & dosage Chalcone / chemistry Chalcone / metabolism Dose-Response Relationship, Drug Drug Delivery Systems / methods* Humans Inflammation / chemically induced Inflammation / metabolism Inflammation / prevention & control Inflammation Mediators / antagonists & inhibitors Inflammation Mediators / metabolism* Lipopolysaccharides / toxicity* Lymphocyte Antigen 96 / metabolism* Male Mice Mice, Inbred C57BL Mice, Knockout Protein Structure, Secondary Protein Structure, Tertiary Sepsis / metabolism* Sepsis / prevention & control*
IF 7.73
引用数 37
WOS 分野 PHARMACOLOGY & PHARMACY
リソース情報
実験動物マウス RBRC02388