抗中性粒细胞胞质-髓过氧化物酶抗体相关性血管炎肽酰基精氨酸脱亚胺酶4表达的变化及临床意义  

A preliminary study on the change and clinical significance of peptidylarginine deiminase 4 expression on the neutrophils in the peripheral blood from the patients with anti-neutrophil cytoplasmic myeloperoxidase antibody-associated vasculitis

作　　者：庞博文 王森 何倩[1] 郑美娟[2] 张铭明[1] 帅宗文[1] Pang Bowen;Wang Sen;He Qian;Zheng Meijuan;Zhang Mingming;Shuai Zongwen(Department of Rheumatology and Immunology,the First Affiliated Hospital of Anhui Medical University.Anhui Province Key Laboratory of Major Autoimmune Diseases,Hefei 230022,China;Flow Cytometry Laboratory,Department of Clinical Laboratory Medicine,First Affiliated Hospital of Anhui Medical Universitythe,Hefei 230022,China)

机构地区：[1]安徽医科大学附属第一医院风湿免疫科,合肥230022 [2]安徽医科大学附属第一医院临床检验医学科流式细胞检测室,合肥230022

出　　处：《中华风湿病学杂志》2020年第8期536-540,共5页Chinese Journal of Rheumatology

基　　金：安徽省重点研究与开发计划项目(1804h08020228)。

摘　　要：目的初步探讨抗中性粒细胞胞质髓过氧化物酶抗体相关性血管炎(MPO-AAV)患者中性粒细胞肽酰基精氨酸脱亚胺酶4(PAD4)表达的变化及临床意义。方法选择39例初诊未治的MPO-AAV患者为病例组,记录患者伯明翰血管炎活动度积分(BVAS),选择39名健康志愿者为对照组。采用流式细胞术检测2组受试者外周血中性粒细胞PAD4的表达水平,ELISA法检测外周血中性粒细胞胞外陷阱(NETs)、补体C5活化片段a(C5a)及抗中性粒细胞胞质髓过氧化物酶抗体(MPO-ANCA)水平。采用t检验比较2组间PAD4、C5a及NETs水平差异,采用Spearman相关分析和多元线性回归分析病例组内各检测指标与BVAS间关系。结果病例组表达PAD4中性粒细胞比例及PAD4表达的平均荧光强度(MIF)均显著高于对照组[分别为(71±11)%与(26±6)%,t=22.456,P<0.01;(33±4)与(14±4),t=18.668,P<0.01]病例组NETs和C5a水平均高于对照组[分别为(0.62±0.22)与(0.26±0.15),t=8.466,P<0.01;(4.6±1.0)ng/ml与(2.9±1.0)ng/ml,t=7.697,P<0.01]。相关分析结果显示病例组BVAS分别与表达PAD4的中性粒细胞百分比、PAD4的MFI、NETs、C5a和MPO-ANCA呈正相关(分别为r=0.843,P<0.01;r=0.821,P<0.01;r=0.4111,P<0.01;r=0.613,P<0.01;r=0.790,P<0.01)。多元线性回归分析显示表达PAD4中性粒细胞百分比和MPO-ANCA水平分别是BVAS的独立影响因素(分别为β=0.325,t=5.889,P<0.01;β=0.796,t=4.298,P<0.01)。结论活动期MPO-AAV患者外周血中性粒细胞PDA4表达水平显著增强,且是影响MPO-AAV疾病活动度的独立因素,干预中性粒细胞PAD4的表达可能是MPO-AAV治疗的潜在新靶点。Objective To investigate the change of peptidylarginine deiminase 4(PAD4)expression in the neutrophils of peripheral blood of patients with anti-neutrophil cytoplasmic myeloperoxidase antibody-associated vasculitis(MPO-AAV),and to analyze the relationship between the change and disease activity.Methods Thirty-nine untreated patients with active MPO-AAV(patient group)and thirty-nine healthy volunteers(control group)were enrolled into this study.The PAD4 expressed on neutrophils was detected by flow cytometry(FCM).The serum neutrophil extracellular traps(NETs),fragments from the activated complement C5(C5a)and anti-neutrophil cytoplasmic myeloperoxidase antibody(MPO-ANCA)were measured by enzyme-linked immuno sorbent assay(ELISA).Their disease activity was evaluated by Birmingham vasculitis activity score(BVAS).All the detected results were compared between the 2 groups by t test,Spearman correlation analysis and multivariate linear regression analysis were employed to analyze the relationship between BVAS and the Lab test results in patient group.Results The proportion of neutrophil expressing PAD4,the mean fluorescence intensity(MIF)of PAD4,the levels of NETs and C5a in patient group were significantly higher than those in the control group[(71±11)%vs(26±6)%,t=22.456,P<0.01;(33±4)vs(14±4),t=18.668,P<0.01;(0.62±0.22)vs(0.26±0.15),t=8.466,P<0.01 and(4.6±1.0)vs(2.9±1.0),t=7.697,P<0.01,respectively].In patient group,BVAS was positively associated with the proportion of PAD4+neutrophil,MFI of PAD4,the serum level of NETs,C5a and MPO-ANCA(r=0.843,P<0.01;r=0.821,P<0.01;r=0.4111,P<0.01;r=0.613,P<0.01 and P=0.790,P<0.01,respectively),however,the results of multiple linear regression analysis showed only the percentage of PAD4+neutrophils and the level of MPO-ANCA were independent influencing factors on BVAS(β=0.3246,P<0.01 andβ=0.796,P<0.01,respectively).Conclusion The percentage of neutrophils expressed PAD4 in the peripheral blood of patient with active MPO-AAV is significantly increased,and it is an independen

关 键 词：肽酰基精氨酸脱亚胺酶4 抗中性粒细胞胞质抗体相关性血管炎 补体C5A 中性粒细胞胞外陷阱 

分 类 号：R593.2[医药卫生—内科学]