中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2014年
7期
931-936
,共6页
Vam3%巨噬细胞%ATP%P2X7%炎症%Ca2+
Vam3%巨噬細胞%ATP%P2X7%炎癥%Ca2+
Vam3%거서세포%ATP%P2X7%염증%Ca2+
Vam3%macrophage%ATP%P2X7%in-flammation%Ca2+
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。
目的：研究二苯乙烯化閤物Vam3對外源性三燐痠腺苷（adenosine triphosphate，ATP）誘導的小鼠腹腔巨噬細胞炎癥反應的影響及機製。方法在預先以脂多糖（lipopolysac-charide，LPS）刺激的小鼠腹腔巨噬細胞培養液中加入 ATP作為體外炎癥模型。ELISA法檢測細胞上清中IL-1β水平；酶活力法檢測 caspase 1酶活力；MTT 法檢測細胞增殖；DCFH-DA（2，7′-dichlorofluorescin diacetate）熒光探針法檢測細胞內活性氧（reactive oxygen species，ROS）水平；激光共聚焦顯微鏡檢測細胞內 Ca2＋濃度。結果 Vam3對外源性ATP誘導的小鼠腹腔巨噬細胞IL-1β分泌增加、caspase 1酶活力增彊、細胞內ROS水平增加等炎癥反應均顯示齣明顯抑製作用，併可明顯降低外源性ATP誘導的小鼠腹腔巨噬細胞毒性及Ca2＋內流。結論 Vam3可能通過榦預caspase 1～IL-1β炎性通路抑製外源性ATP誘導的小鼠腹腔巨噬細胞炎癥反應。
목적：연구이분을희화합물Vam3대외원성삼린산선감（adenosine triphosphate，ATP）유도적소서복강거서세포염증반응적영향급궤제。방법재예선이지다당（lipopolysac-charide，LPS）자격적소서복강거서세포배양액중가입 ATP작위체외염증모형。ELISA법검측세포상청중IL-1β수평；매활역법검측 caspase 1매활력；MTT 법검측세포증식；DCFH-DA（2，7′-dichlorofluorescin diacetate）형광탐침법검측세포내활성양（reactive oxygen species，ROS）수평；격광공취초현미경검측세포내 Ca2＋농도。결과 Vam3대외원성ATP유도적소서복강거서세포IL-1β분비증가、caspase 1매활력증강、세포내ROS수평증가등염증반응균현시출명현억제작용，병가명현강저외원성ATP유도적소서복강거서세포독성급Ca2＋내류。결론 Vam3가능통과간예caspase 1～IL-1β염성통로억제외원성ATP유도적소서복강거서세포염증반응。
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.
