类风湿性关节炎患者血浆和关节滑液中miR-16的表达  

Expression of miR-16 in plasma and synovial fluid of patients with rheumatoid arthritis

作　　者：冯忠军[1] 冯彦蕊[1] 闻海丰[1] 孙寅[2] 

机构地区：[1]河北医科大学第三医院检验科,河北石家庄050051 [2]济宁医学院附属医院检验科,山东济宁272029

出　　处：《河北医科大学学报》2016年第3期296-300,共5页Journal of Hebei Medical University

摘　　要：目的通过检测miR^(-1)6在类风湿关节炎(rheumatoid arthritis,RA)患者血浆和关节滑液中的表达,探讨其在RA鉴别诊断以及病情评估中的应用价值。方法应用茎环逆转录引物的SYBR GreenⅠ实时荧光定量PCR法检测RA患者血浆、关节滑液以及单核细胞(peripheral blood mononuclear cell,PBMC)中miR^(-1)6的表达水平,以U6snRNA为内参,另有10例骨关节炎(osteoarthritis,OA)作病例对照。结果目的基因miR^(-1)6和内参U6snRNA的扩增曲线及熔解曲线均良好,未出现非特异性产物及引物二聚体现象。miR^(-1)6在血浆中表达水平显著高于关节滑液和外周血PBMC(P<0.01);miR^(-1)6在RA患者PBMC中的表达水平显著高于正常对照组(P<0.01)。与OA患者相比,miR^(-1)6在RA患者关节滑液中的表达水平显著升高(P<0.01)。血浆中miR^(-1)6表达与疾病活动度量表(Disease Activity Score,DAS28)评分呈负相关(P<0.05),与红细胞沉降率(erythrocyte sedimentation rate,ESR)及C反应蛋白(C-reactive protein,CRP)均无相关性(P>0.05);PBMC中miR^(-1)6表达与DAS28评分、ESR及CRP均呈正相关(P<0.05);关节滑液中miR^(-1)6表达与DAS28评分、ESR及CRP均无相关性(P>0.05)。结论茎环逆转录引物的SYBR GreenⅠ实时荧光定量PCR法是灵敏而且特异的miRNA检测方法。血浆和关节滑液miR^(-1)6的来源并不相同,在血浆和关节滑液中可能存在不同的miRNA表达谱。miR^(-1)6的表达水平可望用于RA与OA的鉴别诊断以及作为RA病情活动度的有效评价指标。Objective To observe the expression levels of miR^-16 in plasma and synovial fluid from patients with rheumatoid arthritis（RA）,and to explore its value in early differented diagnosis and in prediction of disease severity of RA.Methods Plasma,peripheral blood mononuclear cell（PBMCs）and synovial fluid were taken from patients with RA and osteoarthritis（OA）.The expression level of miR^-16 was detected by stem-loop real-time fluorescent quantitative polymerase chain reaction（stem-loop FQ-PCR）with SYBR GreenⅠdye,and using U6 snRNA as endogenous control.Results Both of the amplication curves and the dissociation curves of miR^-16 and U6snRNA were running well.No primer dimmers were formed,and the products of PCR were specific.The expression level of miR^-16 in plasma was higher than those in synovial fluid and PBMCs,respectively（P〈0.01）.The expression level of miR^-16 in PBMCs was higher for RA patients than for healthy controls（P〈0.01）,the level in synovial fluid was higher for RA patients than for OA patients（P〈0.01）.The expression level of miR^-16 in plasma was inversely correlated with the Disease Activity Score（DAS28）（P〈0.05）,but was not correlated with erythrocyte sedimentation rate（ESR）and C-reactive protein（CRP）.the miR^-16 level in PBMCs was positively correlated with DAS28,ESR and CRP（P〈0.05）.The miR^-16 level in synovial fluid had no correlations with DAS28,ESR and CRP（P〈0.05）.ConclusionStem-loop FQ-PCR with SYBR Green Ⅰ dye was a sensitive and specific method for the quantitative detection of miRNAs.There were distinct profiles in plasma and synovial fluid,and the expression level of miR^-16 in synovial fluid can be a marker for differential diagnosis of RA and OA.The expression level of plasma miR^-16 can serve as an effective novel indicator on assessment of the clinical disease activity.

关 键 词：关节炎 类风湿 滑液 微RNAS 

分 类 号：R593.22[医药卫生—内科学]