天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (2): 146-148.doi: 10.11958/20150240

• 细胞与分子生物学 • 上一篇    下一篇

血清淀粉样蛋白A诱导中性粒细胞胞外诱捕网形成

苏慧慧 1， 万春友 2， 魏蔚 3， 孟海妹 1， 焦亚冲 1， 邢冬红 1， 郑芳 1△   

  1. 1天津医科大学医学检验学院免疫学教研室 （邮编300203）； 2天津市天津医院； 3天津医科大学总医院
  • 收稿日期:2015-10-19 修回日期:2015-11-01 出版日期:2016-02-15 发布日期:2016-02-15
  • 通讯作者: △通讯作者 E-mail： zhengfang@tijmu.edu.cn E-mail:zhengfang129@163.com
  • 基金资助:
    天津市应用基础与前沿技术研究计划项目 （14JCYBJC25600）

Serum amyloid A induces the formation of NETs

SU Huihui1, WAN Chunyou2, WEI Wei3, MENG Haimei1, JIAO Yachong1, XING Donghong, ZHENG Fang1△   

  1. 1 Department of Immunology, School of Laboratory Medicine, Tianjin Medical University, Tianjin 300203, China; 2 Tianjin Hospital; 3 General Hospital of Tianjin Medical University
  • Received:2015-10-19 Revised:2015-11-01 Published:2016-02-15 Online:2016-02-15
  • Contact: △Corresponding Author E-mail: zhengfang@tijmu.edu.cn E-mail:zhengfang129@163.com

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摘要： 目的 探究血清淀粉样蛋白 A（SAA）能否在体外诱导中性粒细胞形成中性粒细胞胞外诱捕网（NETs）。方法 建立稳定的体外诱导 NETs 形成方法， 包括外周血中性粒细胞的分离、 细胞体外培养、 NETs 的诱导和形态学观察。提取健康志愿者的外周血中性粒细胞， 分为阴性对照 （NC） 组、 SAA 组和脂多糖 （LPS） 组， 给予相应的刺激后，观察 NETs 的形成并计算百分率， 酶联免疫吸附试验（ELISA）检测培养上清中人组蛋白 3（h3）的含量。结果 提取的外周血中性粒细胞纯度与细胞活力均在 95%以上。显微镜下观察到部分细胞的细胞核失去原有核型， 向外扩散形成网状， 即 NETs。与 NC 组相比， 接受 SAA 刺激的中性粒细胞有更多的 NETs 形成（P < 0.05）。培养液上清中的 h3 含量显著高于 NC 组 （P < 0.05）。结论 SAA 可在体外诱导中性粒细胞形成 NETs。

关键词: 血清淀粉样蛋白 A, 中性粒细胞胞外诱捕网, 中性粒细胞, 炎症, 人组蛋白 3

Abstract: Objective To explore whether serum amyloid A (SAA) can induce the formation of neutrophil extracellular traps（NETs）in neutrophils in vitro. Methods A stable method for inducing NETs formation in vitro was established, including isolation of peripheral blood neutrophils, cell culture, and NETs formation and observation. The neutrophils were iso⁃ lated from peripheral blood of healthy volunteers. And cells were cultured in vitro and classified into three groups: negative control (NC) group, SAA group and lipopolysaccharide (LPS) group. Following the distinct stimulation in three groups, NETs formation was observed and its percentage was calculated. The concentration of hinstone (h) 3 in supernatant was detected by ELISA. Results The purification and vitality of isolated neutrophils were both more than 95%. The nuclei of neutrophils lost their shape and spread, NETs formation was found. More NETs formation was found in SAA group than that in NC group (P < 0.05). Moreover, the concentration of h3 in supernatant was significantly higher in SAA group than that in NC group (P < 0.05). Conclusion SAA can induce the formation of NETs in vitro.

Key words: serum amyloid A protein, neutrophil extracellular traps, neutrophil, inflammation, hinstone 3