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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
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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 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.
出处 《中国药理学通报》 CAS CSCD 北大核心 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)
关键词 Vam3 巨噬细胞 ATP P2X7 炎症 CA2+ Vam3 macrophage ATP P2X7 in-flammation Ca2+
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