干扰素-λ1/白介素-29抑制脓毒症患者血清C5a诱导的中性粒细胞胞外诱捕网相关的凝血酶生成  

Interferon-λ1/interleukin-29 inhibits neutrophil extracellular traps-associated thrombin production following induced by serum-derived C5a of sepsis patients

作　　者：俞秋兴[1] 杜鸿[1] 张海方[1] 张勇[2] Yu Qiuxing;Du Hong;Zhang Haifang;Zhang Yong(Department of Laboratory Medicine,the Second Clinical Medical School of Soochow University,Suzhou 215004,China;Department of Emergency and Critical Care Medicine,the Second Affiliated Hospital of Soochow University,Suzhou 215004,China)

机构地区：[1]苏州大学第二临床医学院医学检验系,215004 [2]苏州大学附属第二医院急重症医学科,215004

出　　处：《中华急诊医学杂志》2020年第11期1439-1444,共6页Chinese Journal of Emergency Medicine

摘　　要：目的探讨脓毒症血栓形成的炎症环境中血清来源C5a补体能否激活多型核中性粒细胞(polymorphonuclear neutrophils,PMNs)释放中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs),以及干扰素-λ1/白介素29(interferon-λ1/interleukin 29,IFN-λ1/IL-29)是否具有抑制NETs相关凝血酶生成的作用。方法健康志愿者和脓毒症患者的外周血样分别来自2018—2019年苏州大学附属第二医院门诊和住院部,体外研究脓毒症患者血清C5a刺激的健康志愿者外周血分离的PMNs细胞,IL-29和抗CD88单抗作为NETs释放调节因子,免疫荧光分析(immunofluorescence,IF)PMNs上NETs的释放水平,免疫印迹技术(immuno blot,IB)评估PMNs上组织因子(tissue factor,TF)的表达,荧光光谱法(fluorescence spectroscopy,FS)测定上清液中循环DNA水平,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定凝血酶-抗凝血酶(thrombin-antithrombin,TAT)复合物水平。计量资料多组间比较采用单因素方差分析,以P<0.05为差异有统计学意义。结果以脓毒症患者血清C5a诱导的健康志愿者PMNs释放的NETs表达水平(20.5±3.7)%为对照组,IL-29和抗CD88单抗抑制了脓毒症患者血清C5a诱导的健康志愿者PMNs上TF表达的NETs释放,采用IF技术测定的表达水平分别为(5.0±1.5)%、(7.3±3.1)%,差异均有统计学意义(q=9.23和8.73,均P<0.01)。IL-29和抗CD88单抗抑制了脓毒症患者血清C5a诱导的健康志愿者PMNs上TF水平,IB测定的TF表达水平分别为(0.52±0.03)%、(0.57±0.02)%,与脓毒症患者血清C5a诱导的健康志愿者PMNs的TF表达水平(0.81±0.03)%进行比较,差异均有统计学意义(q=3.44和3.21,均P<0.05)。IL-29和抗CD88单抗降低了NETs相关的TAT复合物的水平,分别为(3.3±0.4)ng/mL、(4.6±0.7)ng/mL,与脓毒症患者血清C5a诱导的健康志愿者PMNs释放的NETs相关的TAT复合物水平(7.8±0.7)ng/mL比较,差异均有统计学意义(q=6.77和5.27,均P<0.01)。结论脓毒症患者血清来�Objective To investigate whether serum-derived C5a activates polymorphonuclear neutrophils(PMNs)to release neutrophil extracellular traps(NETs)in the inflammatory environment of thrombotic sepsis and whether interferon-λ1/interleukin 29(IFN-λ1/IL-29)can inhibit the NETs-related thrombin production.Methods Peripheral blood samples of healthy volunteers and sepsis patients were collected from outpatient and inpatient of the Second Affiliated Hospital of Soochow University in 2018 and 2019,respectively.In vitro,PMNs isolated from peripheral blood of healthy volunteers were stimulated by serum-derived C5a of sepsis patients.IL-29 and anti-CD88 monoclonal antibodies were used to regulate NETs release.The expression of NETs was analyzed by immunofluorescence(IF),the expression of tissue factor(TF)was evaluated by immunoblotting(IB),and the levels of circulating DNA from supernatant were determined by fluorescent spectrometry(FS),and the levels of thrombin-antithrombin(TAT)complexes were detected by enzyme-linked immunosorbent assay(ELISA).Differences in levels of detected biomarkers between groups were assessed using One way ANOVA,and a P<0.05 was considered to be statistically significant.Results The levels of NETs released from PMNs of healthy volunteers after stimulation with serum samples of sepsis patients were(20.5±3.7)%according to IF and served as the control group.IL-29 and anti-CD88 monoclonal antibodies inhibited the release of NETs expressed by TF on PMNs of healthy volunteers induced by serum-derived C5a of sepsis patients.The expression levels were(5.0±1.5)%and(7.3±3.1)%detected by IF,respectively.The difference was statistically significant(q=9.23 and 8.73,both P<0.01).IL-29 and anti-CD88 monoclonal antibody inhibited the levels of TF on PMNs of healthy volunteers induced by serum-derived C5a of sepsis patients,and the expression levels of TF were(0.52±0.03)%and(0.57±0.02)%,respectively.Compared with the expression level of TF(0.81±0.03)%of PMNs in healthy volunteers induced by serum-derived C5a

关 键 词：白介素29 C5a补体 中性粒细胞胞外诱捕网 凝血酶 

分 类 号：R459.7[医药卫生—急诊医学]