天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (1): 13-16.doi: 10.11958/20161290

• 细胞与分子生物学 • 上一篇    下一篇

中性粒细胞胞外诱捕网促进类风湿关节炎血管新生的初步研究 #br# #br#

孟海妹 1， 焦亚冲 ， 刘宜昕 ， 万春友 ， 邢冬红 ， 马骏 ， 郑芳    

  1. 1 天津医科大学医学检验学院临床免疫学教研室 （邮编 300203）； 2 天津医院关节中心； 3 天津医科大学公共卫生学院卫生统计 学教研室
  • 收稿日期:2016-11-07 修回日期:2016-11-21 出版日期:2017-01-15 发布日期:2017-01-15
  • 通讯作者: 郑芳 E-mail:menglw0825@163.com
  • 基金资助:
    血清淀粉样蛋白A在类风湿关节炎血管新生机制中的作用

A preliminary study on the promotion of angiogenesis by NETs in rheumatoid arthritis

  1. 1 Department of Clinical Immunology, School of Laboratory Medicine, Tianjin Medical University, Tianjin 300203, China; 2 Joint center, Tianjin Hospital; 3 Department of Health Statistics, School of Public Health, Tianjin Medical University
  • Received:2016-11-07 Revised:2016-11-21 Published:2017-01-15 Online:2017-01-15

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摘要： 摘要： 目的 研究中性粒细胞胞外诱捕网 （NETs） 对类风湿关节炎 （RA） 炎症尤其是血管新生的作用。方法 取 RA 及骨关节炎 （OA） 患者的滑膜组织切片行组织免疫荧光染色， 观察 NETs 在关节滑膜中的存在情况。分离健康人 外周血中性粒细胞， 刺激 NETs 形成， 并提取 NETs 作为刺激剂。MTT 评估 NETs 对人脐静脉内皮细胞 （HUVECs） 及 RA 滑膜成纤维细胞（RAFLS）增殖的影响， 分别对 HUVECs 组和 RAFLS 组处理如下： 对照（等体积培养基）； NETs （0.28 mg/L NETs）。细胞划痕实验评估 NETs 对 HUVECs 迁移能力的影响， 分为 3 组： 对照组、 血管内皮生长因子组 （40 μg/L VEGF） 和 NETs 组 （0.28 mg/L NETs）。结果 （1） 与 OA 相比， RA 滑膜组织中有较多的 NETs 存在。（2） 体外 培养的中性粒细胞刺激后可形成 NETs， 提取的 NETs-DNA 浓度为 27.8 mg/ （L·106细胞）。（3） MTT 实验结果显示， 分 别与各自对照组相比， 低浓度 NETs（0.28 mg/L）可以促进 HUVECs（0.499±0.011 vs. 0.393±0.009， P＜0.05）及 RAFLS （0.266±0.007 vs. 0.192±0.007， P＜0.05） 增殖。（4） 划痕实验显示， 低浓度 NETs（0.28 mg/L） 可促进血管内皮细胞迁移。 结论 RA 滑膜组织中有较多的 NETs 存在， 低浓度 NETs 可以促进血管新生。

关键词: 关节炎, 类风湿； 骨关节炎； 滑膜； 成纤维细胞； 内皮细胞； 中性粒细胞胞外诱捕网； 血管新生

Abstract: Abstract: Objective To investigate the effect of neutrophil extracellular traps (NETs) on inflammation of rheumatoid arthritis (RA), especially angiogenesis. Methods The presence of NETs in synovial tissues of RA and osteoarthritis (OA) was observed by immunofluorescence assay. Neutrophils were isolated from peripheral blood of health volunteers. Neutrophils were cultured in vitro, the formation of NETs was observed. NETs were extracted as a stimulating agent. The effects of NETs on the proliferation of human umbilical vein endothelial cells (HUVECs) and synovial fibroblasts (RAFLS) were evaluated by MTT, and which were classified into two groups: HUVECs group and RAFLS group, with the following treatment: control and NETs (0.28 mg/L). Wound repair assay was employed to evaluate the effect on the cell migration stimulated with NETs. The experiment was divided into three groups: control, VEGF (40 μg/L VEGF) and NETs (0.28 mg/L NETs). Results (1) Compared with OA, NETs were found more in the synovial tissue of RA. (2) NETs formation was induced by stimulator in vitro. The concentration of extracted NETs- DNA was 27.8 mg/L. (3) MTT assay showed that compared with the control groups, low concentration of NETs (0.28 mg/L) promoted the proliferation of HUVECs (0.499 ± 0.011 vs. 0.393 ± 0.009, P＜0.05) and RAFLS (0.266 ± 0.007 vs. 0.192 ± 0.007, P＜0.05). (4) It was showed that a significant wound closure induced by low concentration of NETs (0.28 mg/L) was found compared with control. Conclusion Our present study suggests that NETs are found more in the synovial tissue of RA, and low concentration of NETs can promote angiogenesis in RA.

Key words: arthritis, rheumatoid, osteoarthritis, synovial membrane, fibroblasts, endothelial cells, neutrophil
extracellular traps, angiogenesis