摘要
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.
出处
《国际免疫学杂志》
CAS
2016年第4期319-322,共4页
International Journal of Immunology
基金
基金项目：国家自然科学基金（81400790,81061120527,81370445,81472408）
卫生部公益性研究基金（201302008）
国家科技部十二五支撑计划（2012BA110801）
