类风湿关节炎患者血清抗核抗体表达分析

Analysis on expression of serum antinuclear antibody in patients with rheumatoid arthritis

目的探讨抗核抗体(ANA)在类风湿关节炎(RA)患者血清中的表达及应用价值。方法选择德阳市人民医院风湿免疫科155例确诊RA患者作为研究对象(RA组),根据患者休息时关节疼痛和肿胀数是否超过5个、晨僵持续时间是否大于1 h、关节压痛个数是否超过5个以及红细胞沉降率(ESR)水平等情况,将RA患者分为稳定组(66例)和活动组(89例);另选本院120例健康体检者作为健康对照组。检测所有受检者的血清ANA、抗双链DNA(dsDNA)和抗核抗体谱(ANAs),ANA和抗dsDNA抗体采用间接免疫荧光法(IIF);检测ANAs 13项〔包括抗核糖体蛋白/史密斯(U1-nRNP/Sm)抗体、抗史密斯(Sm)抗体、抗干燥综合征抗原A(SSA)抗体、抗干燥综合征抗原B(SSB)抗体、抗Ro-52抗体、抗硬皮病70(Scl-70)抗体、抗组氨酰tRNA合成酶(Jo-1)抗体、抗着丝点B蛋白(CENP-B)抗体、抗核小体(Nucleosome)抗体、抗dsDNA抗体、抗组蛋白(Histone)抗体、抗核糖体P蛋白(Rib-Prot)抗体、抗线粒体M2型(AMA-M2)抗体〕采用线性免疫印迹法(LIA)。比较两组之间ANA阳性率,分析ANA荧光模式分布以及ANAs阳性检出情况。结果RA组ANA总阳性率明显高于健康对照组〔38.06%(59/155)比10.83%(13/120),P<0.05〕,其中RA组男性ANA阳性率与女性比较差异无统计学意义(P>0.05)。59例ANA阳性患者标本中,ANA滴度≥1:100者29例(占49.15%),≥1:320者21例(占35.59%),≥1:1000者9例(占15.25%)。ANA荧光模式以核颗粒型(30.51%)、胞质颗粒型(28.81%)、抗着丝粒型(15.26%)、核仁型(10.17%)和核点型(8.47%)最常见。活动组ANA阳性率明显高于稳定组〔46.07%(41/89)比27.27%(18/66),P<0.05〕。ANAs单项或多项抗体阳性率为47.46%(28/59),检出率较高的单项自身抗体分别为抗Ro-52抗体、抗SSA-60抗体、抗SSB抗体、抗U1-nRNP/Sm抗体和抗AMA-M2抗体。结论RA患者血清ANA和ANAs检出率较高,可能与疾病活跃程度或严重程度相关,应引起临床高度重视。

Objective To investigate the expression and clinical significance of antinuclear antibodies(ANA)in serum of patients with rheumatoid arthritis(RA).Methods Totally 155 patients with RA diagnosed by Rheumatology and Immunology Department of Deyang People’s Hospital were selected as the research objects.According to whether the number of arthralgia and swelling at rest was more than 5,whether the duration of morning stiffness was more than 1 hour,whether the number of joint tenderness exceeded 5,erythrocyte sedimentation rate(ESR)levels,etc.,RA patients were divided into stable group(66 cases)and active group(89 cases).Other 120 healthy subjects in our hospital were selected as the control group.The ANA,anti-double-stranded DNA(dsDNA)and antinuclear antibody profiles(ANAs)in serum of all subjects were detected.ANA and anti-dsDNA antibody were detected by indirect immunofluorescence(IIF).The ANAs 13 items[anti-ribosomal protein/Smith(U1-nRNP/Sm)antibody,anti-Smith(Sm)antibody,anti-Sjogren’s syndrome antigen A(SSA)antibody,anti-Sjogren’s syndrome antigen B(SSB)antibody,anti-Ro-52 antibody,anti-scleroderma 70(Scl-70)antibody,anti-Hylecatyl tRNA synthetase(Jo-1)antibody,anti-central point B protein(CENP-B)antibody,anti-neucleosome(Neucleosome)antibody,anti-dsDNA antibody,anti-histone(Histone)antibody,anti-ribosomal P protein(Rib-Prot)antibody,anti-mitochondrial M2(AMA-M2)antibody]were detected by linear immunoblotting(LIA).The positive rates of ANA in each group were compared,and the distribution of ANA fluorescence pattern and positive detection of ANAs were analyzed.Results The total positive rate of ANA in RA group was significantly higher than that in healthy control group[38.06%(59/155)vs.10.83%(13/120),P<0.05],and there was no statistically significant difference between male and female in RA group(P>0.05).Among the 59 ANA positive patients,29 patients(49.15%)had ANA titer≥1:100,21 patients(35.59%)had ANA titer≥1:320,and 9 patients(15.25%)had ANA titer≥1:1000.The most common patterns of ANA fluorescence were nuclear granule(30.51%),cytoplasmic granule(28.81%),anticentromere(15.25%),nucleolar(10.17%)and nuclear point(8.47%).The positive rate of ANA in active group was significantly higher than that in stable group[46.07%(41/89)vs.27.27%(18/66),P<0.05].The positive rate of single or multiple antibodies in ANAs was 47.46%(28/59).The single autoantibodies with higher detection rate were anti-Ro-52 antibody,anti-SSA-60 antibody,anti-SSB antibody,anti-U1-nRNP/Sm antibody and anti-AMA-M2 antibody.Conclusion The detection rate of ANA and ANAs in serum of RA patients is high,which may be related to the degree of disease activity or severity,so it should be paid more attention to in clinical practice.