二苯乙烯化合物Vam3对ATP诱导的小鼠腹腔巨噬细胞炎症反应的影响及机制研究  

Influence of dihydroxy-stilbene compound Vam3 on ATP-induced inflammatory response in macrophages and the underlying mechanisms

作　　者：曹淑花 姚春所 侯琦 

机构地区：[1]中国医学科学院北京协和医学院药物研究所,天然药物活性物质与功能国家重点实验室,北京100050

出　　处：《中国药理学通报》2014年第7期931-936,共6页Chinese Pharmacological Bulletin

基　　金："国家十二五"综合大平台-子课题"抗炎候选物研究"(No 29018)

摘　　要：目的：研究二苯乙烯化合物Vam3对外源性三磷酸腺苷（adenosine triphosphate，ATP）诱导的小鼠腹腔巨噬细胞炎症反应的影响及机制。方法在预先以脂多糖（lipopolysac-charide，LPS）刺激的小鼠腹腔巨噬细胞培养液中加入 ATP作为体外炎症模型。ELISA法检测细胞上清中IL-1β水平；酶活力法检测 caspase 1酶活力；MTT 法检测细胞增殖；DCFH-DA（2，7′-dichlorofluorescin diacetate）荧光探针法检测细胞内活性氧（reactive oxygen species，ROS）水平；激光共聚焦显微镜检测细胞内 Ca2＋浓度。结果 Vam3对外源性ATP诱导的小鼠腹腔巨噬细胞IL-1β分泌增加、caspase 1酶活力增强、细胞内ROS水平增加等炎症反应均显示出明显抑制作用，并可明显降低外源性ATP诱导的小鼠腹腔巨噬细胞毒性及Ca2＋内流。结论 Vam3可能通过干预caspase 1～IL-1β炎性通路抑制外源性ATP诱导的小鼠腹腔巨噬细胞炎症反应。Aim To investigate the effects of Vam3 on ATP-induced inflammatory response in macrophages and the underlying mechanisms. Methods LPS primed mouse peritoneal macrophages were stimulated with ATP,and IL-1βlevel in supernatants was meas-ured by ELISA.Activity of caspase 1 was measured u-sing caspase 1 activity assay kit.Reactive oxygen spe-cies （ROS ）level was detected with fluorescent probe DCFH-DA.MTT assay was used to detect cell prolifer-ation,and intracellular Ca2＋concentration was meas-ured using laser scanning confocal microscope.Results＆amp;nbsp;Extracellular ATP led to increase in IL-1βrelease, caspase 1 activity and ROS production.It also led to rapid increase in intracellular Ca2＋concentration and induced cell death.These effects were inhibited by Vam3 .Conclusion Vam3 inhibits ATP-induced in-flammatory response in macrophages,which may sug-gest the blocking effect of Vam3 on caspase 1 ~IL-1βinflammatory signaling pathway in macrophages.

关 键 词：Vam3 巨噬细胞 ATP P2X7 炎症 CA2+ 

分 类 号：R332[医药卫生—人体生理学] R329.24[医药卫生—基础医学]