中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2014年
11期
738-742,后插1
,共6页
红斑狼疮,系统性%中性粒细胞胞外网络%线粒体DNA%Ⅰ型干扰素
紅斑狼瘡,繫統性%中性粒細胞胞外網絡%線粒體DNA%Ⅰ型榦擾素
홍반랑창,계통성%중성립세포포외망락%선립체DNA%Ⅰ형간우소
Lupus erythematosus,systemic%Neutrophil extracellular trap%Mitochondrial DNA%Type Ⅰ interferon
目的 探讨中性粒细胞胞外网络(NET)中的线粒体DNA(mtDNA)及抗mtDNA抗体与SLE的临床相关性.方法 102例SLE患者、30例RA患者和40名健康对照纳入本实验.分离外周血中性粒细胞,佛波酯刺激后荧光染色观察NET形成;并对LN肾组织标本原位NET染色.采用实时定量PCR检测SLE患者和健康对照NET中mtDNA含量及5个Ⅰ型IFN诱导基因(OAS-1、Mx-1、Ly6e、IFIT1和IFIT4)的mRNA水平.运用ELISA法检测SLE患者、RA患者和健康对照血浆抗mtDNA抗体的滴度.统计学方法采用t检验、x2检验和Spearman相关分析.结果 SLE患者NET中mtDNA含量(1 088 000±1 133 000)显著高于健康对照组(465 900±447 200)(t=2.617,P＜0.01).SLE患者中度活动组(728 300±100 3000)和重度活动组(1 093 000±946 500)的NET中mtDNA的含量都较轻度活动组(159 500±155 100)显著增高(t=2.240,P＜0.05;t=3.894,P＜0.01),同时NET中mtDNA含量还与IFN积分呈正相关(r=0.460 6,P＜0.01).SLE组血浆抗mtDNA抗体水平(1.280.68)显著高于RA组(0.59±0.18) (P＜0.01)和健康对照组(0.70±0.31)(P＜0.01).SLE患者中度活动组(1.3±0.6)和重度活动组(1.4±0.7)的抗mtDNA抗体水平都较轻度活动组(0.7±0.4)显著增高(t=3.154,3.538,P均＜0.01),同时抗mtDNA抗体水平还与抗dsDNA抗体(r=0.542 9,P＜0.001)及IFN积分(r=0.292 8,P＜0.05)呈正相关.抗mtDNA抗体水平与LN的发生显著相关(x2=8.644,P＜0.01).结论 NET线粒体DNA及其抗体是反映SLE疾病活动,并与活动性LN相关的新的生物标志物.
目的 探討中性粒細胞胞外網絡(NET)中的線粒體DNA(mtDNA)及抗mtDNA抗體與SLE的臨床相關性.方法 102例SLE患者、30例RA患者和40名健康對照納入本實驗.分離外週血中性粒細胞,彿波酯刺激後熒光染色觀察NET形成;併對LN腎組織標本原位NET染色.採用實時定量PCR檢測SLE患者和健康對照NET中mtDNA含量及5箇Ⅰ型IFN誘導基因(OAS-1、Mx-1、Ly6e、IFIT1和IFIT4)的mRNA水平.運用ELISA法檢測SLE患者、RA患者和健康對照血漿抗mtDNA抗體的滴度.統計學方法採用t檢驗、x2檢驗和Spearman相關分析.結果 SLE患者NET中mtDNA含量(1 088 000±1 133 000)顯著高于健康對照組(465 900±447 200)(t=2.617,P＜0.01).SLE患者中度活動組(728 300±100 3000)和重度活動組(1 093 000±946 500)的NET中mtDNA的含量都較輕度活動組(159 500±155 100)顯著增高(t=2.240,P＜0.05;t=3.894,P＜0.01),同時NET中mtDNA含量還與IFN積分呈正相關(r=0.460 6,P＜0.01).SLE組血漿抗mtDNA抗體水平(1.280.68)顯著高于RA組(0.59±0.18) (P＜0.01)和健康對照組(0.70±0.31)(P＜0.01).SLE患者中度活動組(1.3±0.6)和重度活動組(1.4±0.7)的抗mtDNA抗體水平都較輕度活動組(0.7±0.4)顯著增高(t=3.154,3.538,P均＜0.01),同時抗mtDNA抗體水平還與抗dsDNA抗體(r=0.542 9,P＜0.001)及IFN積分(r=0.292 8,P＜0.05)呈正相關.抗mtDNA抗體水平與LN的髮生顯著相關(x2=8.644,P＜0.01).結論 NET線粒體DNA及其抗體是反映SLE疾病活動,併與活動性LN相關的新的生物標誌物.
목적 탐토중성립세포포외망락(NET)중적선립체DNA(mtDNA)급항mtDNA항체여SLE적림상상관성.방법 102례SLE환자、30례RA환자화40명건강대조납입본실험.분리외주혈중성립세포,불파지자격후형광염색관찰NET형성;병대LN신조직표본원위NET염색.채용실시정량PCR검측SLE환자화건강대조NET중mtDNA함량급5개Ⅰ형IFN유도기인(OAS-1、Mx-1、Ly6e、IFIT1화IFIT4)적mRNA수평.운용ELISA법검측SLE환자、RA환자화건강대조혈장항mtDNA항체적적도.통계학방법채용t검험、x2검험화Spearman상관분석.결과 SLE환자NET중mtDNA함량(1 088 000±1 133 000)현저고우건강대조조(465 900±447 200)(t=2.617,P＜0.01).SLE환자중도활동조(728 300±100 3000)화중도활동조(1 093 000±946 500)적NET중mtDNA적함량도교경도활동조(159 500±155 100)현저증고(t=2.240,P＜0.05;t=3.894,P＜0.01),동시NET중mtDNA함량환여IFN적분정정상관(r=0.460 6,P＜0.01).SLE조혈장항mtDNA항체수평(1.280.68)현저고우RA조(0.59±0.18) (P＜0.01)화건강대조조(0.70±0.31)(P＜0.01).SLE환자중도활동조(1.3±0.6)화중도활동조(1.4±0.7)적항mtDNA항체수평도교경도활동조(0.7±0.4)현저증고(t=3.154,3.538,P균＜0.01),동시항mtDNA항체수평환여항dsDNA항체(r=0.542 9,P＜0.001)급IFN적분(r=0.292 8,P＜0.05)정정상관.항mtDNA항체수평여LN적발생현저상관(x2=8.644,P＜0.01).결론 NET선립체DNA급기항체시반영SLE질병활동,병여활동성LN상관적신적생물표지물.
Objective To correlate the neutrophil extracellular trap (NET) mitochondrial DNA (mtDNA) and anti-mtDNA antibodies (Abs) with disease activity and clinical features in systemic lupus erythematosus (SLE) patients.Methods We enrolled 102 SLE patients,rheumatoid arthritis (RA) patients (n=30) and healthy donors as controls (n=40).NET were generated from phorbol 12-myristate 13-acetate (PMA)-stimulated peripheral neutrophils.mtDNA levels and the transcriptional levels of five interferon inducible genes(IFIGs)(OAS-1,Mx-1,Ly6e,IFIT1 and IFIT4) were measured by quantitative PCR.Interferon scores (IFN scores) were calculated.Anti-mtDNA Abs were detected by enzyme-linked immunosorbent assay.Spearman's correlation analysis,t test andx2 test were used for statistical analysis.Results mtDNA release by netting neutrophils was greatly enhanced in SLE patients (1 088 000 ±1 133 000) compared with healthy controls (465 900±447 200)(t=2.617,P＜0.01) and significantly correlated with IFN scores (r=0.460 6,P＜0.01).NETs mtDNA in moderate active group (728 300±1 003 000) and severe active group (1 093 000±946 500) were significantly higher than the mild active group (159 500±155 100) (t=2.240,P＜0.05,t=3.894,P＜0.01).Forty-one percent of SLE patients were positive for anti-mtDNA Abs(1.28±0.68),while none of the healthy donors (0.70±0.31) (P＜0.01) and RA controls (0.59±0.18)(P＜0.01) displayed a positive serology response to mtDNA.Addition-ally,the titers of anti-mtDNA Abs were also associated with IFN scores (r=0.292 8,P＜0.05).Anti-mtDNA Abs in moderate active group (1.3±-0.6) and severe active group(1.4±0.7)were significantly higher than the mild active group (0.7±0.4) (t=3.154,3.538,all P＜0.01).The levels of anti-mtDNA Abs significantly correlated with classic an-ti-dsDNA Abs titers measured by Farr assay (r=0.542 9,P＜0.001) and were associated with LN (x2=8.644,P＜0.01).Conclusion mtDNA in NET and anti-mtDNA Abs serve as new biomarkers for disease activity and renal involvement in SLE patients.
