NETs 刺激下慢性支气管炎大鼠血清ANCA 产生机制探讨

目的:探讨长期慢性感染刺激诱导机体产生抗中性粒细胞胞浆抗体(ANCA)的机制。方法:烟熏法联合脂多糖气管内注入,建立慢性支气管炎大鼠模型,用佛波脂(PMA)体外刺激大鼠中性粒细胞产生中性粒细胞胞外网络(NETs),以NETs 免疫慢性支气管炎大鼠,比较慢性支气管炎NETs 刺激组(CB/ NETs)、慢性支气管炎组(CB)和正常组三组大鼠血清髓过氧化物酶鄄抗中性粒细胞胞浆抗体(MPO-ANCA)、血清瓜氨酸化的组蛋白H3(CitH3)、血清IFN-α水平及血清肌酐、血清尿素氮、尿红细胞计数、24 h 尿蛋白定量水平。结果:(1)CB/ NETs 组的血清MPO-ANCA 水平明显高于CB 组及正常组,(62.497 5±19.637 64)ng/ ml vs(17.380 6±4.886 49)ng/ ml,(20.888 4±4.396 07)ng/ ml(P<0.01)。(2)CB/ NETs 组的血清CitH3 水平明显高于CB 组及正常组,(32.597 4±20.291 5)ng/ ml vs (8.856 8±2.366 92)ng/ ml,(5.622 5±0.837 38)ng/ ml,(P<0.05,P<0.01)。(3)CB/ NETs 组中有两只大鼠的血清IFN-α水平异常升高(3 735.7、3 428.4 pg/ ml)。(4)三组大鼠的肾脏相关指标差异暂未呈现出统计学意义。(5)大鼠血清CitH3 水平与血清MPO-ANCA 水平呈正相关(r =0.61,P<0.05)。结论:慢性支气管炎长期、反复感染可导致ANCA 水平升高,从形成高水平的ANCA 到最终形成ANCA 相关性小血管炎尚需要时间。

Neutrophil extracellular traps induce production of MPO-ANCA in chronic bronchitis rats

Objective:To investigate the mechanism of chronic bronchitis with recurrent long duration infections in the production of MPO anti-neutrophil cytoplasmic antibodies(ANCA).Methods:Rats were gave intratracheal injection of LPS and smoked to established chronic bronchitis(CB) model.Rat neutrophils were incubated with phorbol-12-myristate-13-acetate(PMA) to induce Neutrophil extracellular traps(NETs).NETs were then used to immune chronic bronchitis rats.Serum concentrations of myeloperoxidase anti-neutrophil cytoplasmic antibodies(MPO-ANCA),Citrullinated Histone H3(CitH3),Interferon-α(IFN-α)were evaluated.Results:(1)The serum levels of MPO-ANCA in chronic bronchitis rats immuned with NETs(CB/ NETs)were significantly higher than those in chronic bronchitis rats(CB)and healthy control rats,(62.497 5±19.637 64)ng/ ml vs (17.380 6±4.886 49)ng/ ml,(20.888 4±4.396 07)ng/ ml(both P<0.01).(2)The serum levels of CitH3 in CB/ NETs rat were significantly higher than those in CB rat and healthy control rats,(32.597 4±20.291 5)ng/ ml vs (8.856 8±2.366 92)ng/ ml,(5.622 5±0.837 38)ng/ ml(P<0.05,P<0.01).(3)The serum levels of IFN-αwere found increased sharply in two CB/ NETs rats(3 735.7,3 428.4 pg/ ml).(4) Renal function among three groups showed no significant difference yet.(5)Serum CitH3 level showed a positive correlation with serum MPO-ANCA level(r =0.61,P<0.05).Conclusion:Chronic bronchitis with recurrent infections could induce the production of ANCA probably by the stimulation of NETs,but whether it might develop to AAV remains need a duration.