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目的 观察curdlan刺激下巨噬细胞活性氧(ROS)生成的变化及Ca2对活性氧生成的影响,探讨Ca2+参与curdlan诱导的巨噬细胞炎症反应的可能机制.方法 将Dectin-1缺失的小鼠和正对照WT小鼠来源的巨噬细胞,以100 μg/mL curdlan处理从上述小鼠中分离培养的巨噬细胞,建立巨噬细胞炎症反应模型.处理后,Amplex Red和luminol法分别检测巨噬细胞中超氧离子和过氧化氢的生成变化.用EGTA螯合检测试剂中Ca2+,观察无Ca2情况下活性氧的产生情况,以及在检测试剂中加入不同浓度的Ca2+观察其对ROS产生的影响.结果 Curdlan刺激下,巨噬细胞(骨髓来源和腹腔来源巨噬细胞)中活性氧(包括超氧离子和过氧化氢)水平显著增加.与无Ca2组相比(0 mmol/L),随着Ca2+浓度增加(1.5、2.5 mmol/L),curdlan诱导的巨噬细胞中ROS水平升高.结论 Curdlan能够结合巨噬细胞中相应的免疫识别受体(dectin-1)通过产生大量ROS从而介导炎症反应,调控ROS的水平可能是Ca2+参与炎症反应的机制之一.“,”Objective To investigate reactive oxygen species(ROS) production from macrophages induced by curdlan and to study the effects of Ca2+ on ROS production.Methods Macrophages (BMDM and peritoneal macrophages) were collected from (wild type,WT) mice and Dectin-1 (kick out,KO) mice,and cells they were stimulated by curdlan (100 μg/ mL) to establish cell model.ROS production including super oxide and H2O2 was estimated by luminol and Amplex Red,respectively.Then Ca2 + was eliminated by EGTA.ROS production was evaluated with EGTA or without EGTA but with various concentrations of Ca2 +.Results Increased ROS production was seen in macrophages (BMDM and peritoneal macrophages).ROS production induced by curdlan increased with the increased Ca2 + concentration (1.5,2.5mmol/L).Conclusions Curdlan can activate production of ROS through dectin-1 recognition.Ca2+ might be involved in inflammatory reaction via regulating the ROS production.