循环microRNAs与局灶节段性肾小球硬化患者疾病活动的关系

Circulating microRNAs and the disease activity of patients with focal segmental glomerulosclerosis

目的:分析循环中与局灶节段性肾小球硬化(FSGS)患者疾病活动相关microRNAs(miRNAs),为后续探讨循环miRNAs在FSGS发病中的作用奠定基础。方法:选取年龄、性别匹配的活动性FSGS患者及正常对照各9例,提取FSGS患者及对照组血浆总RNA,采用TaqMan低密度芯片(TaqMan Low Density Array)进行miRNA表达谱分析,筛选出在活动性FSGS患者循环中表达上调的miRNAs分子;然后再选取32例活动性FSGS患者血浆,采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)方法对筛选出的miRNAs分子进行验证,同时与非活动性FSGS患者进行比较。采用ROC曲线分析循环miRNA区分活动性FSGS、非活动性FSGS和正常对照的效能。结果:与正常对照相比,活动性FSGS患者循环miRNA-125b,miRNA-186和miRNA-193a-3p水平显著上调(P<0.05)。ROC曲线分析显示,循环miRNA-125b、miRNA-186和miRNA-193a-3p区分活动性FSGS与正常对照的ROC曲线下面积(AUC)分别为0.882、0.789和0.910。与非活动性FSGS相比,活动性FSGS患者循环中miRNA-186水平也明显上调。结论:活动性FSGS患者循环miRNA-125b、miRNA-186及miRNA-193a-3p较正常对照显著升高,其中miRNA-186的升高与疾病的活动性关系更密切。

Objective：To analyze the elationship between circulating miRNAs and the disease activity of patients with FSGS,and explore the role of circulating miRNAs in the pathogenesis of FSGS.Methodology：miRNAs expression profile was performed on 2 pooled plasma samples from nine patients with active FSGS and 9 normal controls using TaqMan Low Density Array.A panel of differential expressed miRNAs were found,and then validated by quantitative reverse transcription PCR （qRT-PCR） array with individual plasma samples.Meanwhile,plasma samples from non-active FSGS patients were examined for candidate miRNAs.Receiver-operatin characteristic （ROC） curves were established to analyze the value of circulating miRNAs for differentiating active FSGS from non-active FSGS and healthy controls.Results：The results of TaqMan Low Density Array indicated that a panel of miRNAs was deregulated in plasma of active FSGS patients.After large scale independent cohort validation by qRT-PCR,the levels of circulating miRNA-125b,miRNA-186 and miRNA-193a-3p were significantly increased in active FSGS patients compared with that in normal controls.ROC curve analysis showed that circulating miRNA-125b,miRNA-186 and miRNA-193a-3p was a significant value for differentiating active FSGS from normal controls.In addition,the level of circulating miRNA-186 was also significantly increased in active FSGS patients when compared with non-active FSGS patients.Conclusion：We determined that the levels of circulating miRNA-125b,miRNA-186 and miRNA-193a-3p were increased in patients with active FSGS and miR-186 showed a more closer relationship with FSGS activity.