【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(40μg/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。更多还原

【Abstract】 Objective To investigate the effect of neutrophil extracellular traps(NETs) on inflammation of rheumatoidarthritis(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. Theeffects 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 followingtreatment: control and NETs(0.28 mg/L). Wound repair assay was employed to evaluate the effect on the cell migrationstimulated with NETs. The experiment was divided into three groups: control, VEGF(40 μg/L VEGF) and NETs(0.28 mg/LNETs). Results(1) Compared with OA, NETs were found more in the synovial tissue of RA.(2) NETs formation wasinduced by stimulator in vitro. The concentration of extracted NETs- DNA was 27.8 mg/L.(3) MTT assay showed thatcompared 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 asignificant 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 canpromote angiogenesis in RA.更多还原