Wang Huan, Sha Li-Li, Ma Tian-Tian, Zhang Lu-Xia, Chen Min, Zhao Ming-Hui
Renal Division, Department of Medicine, Peking University, First Hospital, Beijing 100034, China.
Peking University Institute of Nephrology, Beijing 100034, China.
PLoS One. 2016 Feb 3;11(2):e0148197. doi: 10.1371/journal.pone.0148197. eCollection 2016.
Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a group of life-threatening disorders, and frequently affects the kidneys. This study investigated whether the circulating neutrophil extracellular traps (NETs) levels were associated with disease activity of AAV. We collected serum samples from 34 patients with AAV in active stage and 62 patients with AAV in remission. Cell free DNA in serum was quantified using the Quant-iT PicoGreen assay. NETs associated MPO-DNA complexes, citrullinated-histone H3-DNA (cit-H3-DNA) complexes and the concentration of deoxyribonuclease I (DNase I) were quantified using ELISA. The activity of DNase I was quantified using radial enzyme-diffusion method. Associations between circulating levels of NETs with clinico-pathological parameters were analyzed. Serum levels of NETs in active AAV patients were significantly higher than those in healthy controls, and the level of cell free DNA correlated with C-reactive protein (CRP). However, no correlation was found between MPO-DNA complexes or cit-H3-DNA complexes level and CRP. Also there was no significant correlation between NETs level and initial serum creatinine, estimated glomerular filtration rate (eGFR), crescents formation or Birmingham Vasculitis Activity Score (BVAS). Furthermore, there was no significant difference of serum levels of cell free DNA or MPO-DNA complexes between active stage and remission of AAV. In conclusion, circulating levels of NETs cannot be used as a biomarker to assess disease activity in AAV patients.
抗中性粒细胞胞浆抗体(ANCA)相关性血管炎(AAV)是一组危及生命的疾病,常累及肾脏。本研究调查了循环中性粒细胞胞外陷阱(NETs)水平是否与AAV的疾病活动相关。我们收集了34例活动期AAV患者和62例缓解期AAV患者的血清样本。采用Quant-iT PicoGreen分析法对血清中的游离DNA进行定量。使用酶联免疫吸附测定法(ELISA)对与NETs相关的髓过氧化物酶-DNA复合物、瓜氨酸化组蛋白H3-DNA(cit-H3-DNA)复合物以及脱氧核糖核酸酶I(DNase I)的浓度进行定量。采用放射酶扩散法对DNase I的活性进行定量。分析了NETs循环水平与临床病理参数之间的关联。活动期AAV患者的血清NETs水平显著高于健康对照,且游离DNA水平与C反应蛋白(CRP)相关。然而,未发现MPO-DNA复合物或cit-H3-DNA复合物水平与CRP之间存在相关性。此外,NETs水平与初始血清肌酐、估计肾小球滤过率(eGFR)、新月体形成或伯明翰血管炎活动评分(BVAS)之间也无显著相关性。此外,AAV活动期和缓解期之间血清游离DNA或MPO-DNA复合物水平无显著差异。总之,循环NETs水平不能用作评估AAV患者疾病活动的生物标志物。