瑞香素对HMGB1释放及HMGB1诱发的炎症反应的双重抑制作用  

Dual role of daphnetin in suppressing HMGB1 release and HMGB1-induced inflammation in murine macrophage RAW264.7 cells and human monocytic THP-1 cells in vitro

作　　者：戚之琳[1,2] 齐世美[1,2] 凌烈锋[1,2] 冯遵永[2] 

机构地区：[1]皖南医学院生物化学教研室 [2]安徽省活性生物大分子重点实验室,安徽芜湖241002

出　　处：《南方医科大学学报》2015年第11期1519-1523,共5页Journal of Southern Medical University

基　　金：国家自然科学基金青年项目(31301171);安徽省教育厅自然科学研究项目(KJ2013B311);皖南医学院校重点科研项目培育基金(WK2015Z01);皖南医学院博士启动基金~~

摘　　要：目的探讨瑞香素对高迁移率族蛋白1(HMGB1)释放及其诱发炎症反应的双重抑制作用。方法 RAW264.7细胞常规培养,不同浓度的瑞香素单独或联合脂多糖(LPS)作用细胞不同的时间,ELISA检测晚期炎症因子HMGB1的释放;Western blot检测JAK1/2、STAT1的磷酸化。THP-1细胞常规培养,不同浓度的瑞香素单独或联合rh HMGB1刺激细胞不同的时间,ELISA检测炎症因子TNF-α,IL-6,PGE2的释放;NO检测试剂盒检测NO水平;Western blot检测i NOS、COX-2的表达及p38、ERK、JNK的磷酸化水平。结果瑞香素浓度依赖性的下调HMGB1的释放,抑制rh HMGB1诱导的i NOS、COX-2的表达及TNF-α,IL-6,PGE2、NO的释放。WB结果显示,瑞香素显著下调脂多糖诱导的JAK-STAT1信号磷酸化,但对rh HMGB1诱导的MAPKs磷酸化无明显抑制作用。结论瑞香素能够抑制HMGB1释放及其诱发的炎症反应,而且瑞香素可能通过抑制JAK-STAT1信号途径减弱HMGB1的释放。Objective To investigate the dual role of daphnetin in suppressing high mobility group box-1 protein（HMGB1）release and blocking HMGB1-induced inflammatory response. Methods Murine macrophage RAW264.7 cells were cultured in the presence of daphnetin, lipopolysaccharide（LPS）, or both. HMGB1 release from the cells was determined using ELISA, and phosphorylations of JAK1/2 and of STAT1 were detected by Western blotting. Human monocytic THP-1 cells exposed to daphnetin, rh HMGB1, or both were examined for NO production using a NO detection kit, for the release of tumor necrosis factor-α（TNF-α）, interleukin-6（IL-6）, and prostaglandin E2（PGE2） using ELISA, and for expressions of i NOS, COX-2 and phosphorylated p38, ERK, and JNK with Western blotting. Results Daphnetin dose-dependently reduced the release of HMGB1 in RAW264.7 cells and suppressed rh HMGB1-induced i NOS and COX-2 expressions and release of TNF-α, IL-6,PGE2, and NO in THP-1 cells. Western blotting revealed that daphnetin significantly down-regulated the phosphorylations of JAK-STAT1 pathway in LPS-stimulated RAW264.7 cells but did not suppress the phosphorylations of MAPKs signaling pathway induced by rh HMGB1 in THP-1 cells. Conclusion Daphnetin can reduce the release of HMGB1 and suppress HMGB1-induced inflammatory response. In RAW264.7 cells, daphnetin inhibited LPS induced HMGB1 release is at least partly mediated by suppressing JAK-STAT1 signaling pathway activation.

关 键 词：瑞香素 HMGB1 脂多糖 炎症 

分 类 号：R285[医药卫生—中药学]