小儿原发性肾病综合征患者血清和尿液miR-30a-5p水平增加及临床意义  

Elevated Serum and Urinary miR-30a-5p Level in Children with Idiopathic Nephrotic Syndrome and Its Clinical Value

作　　者：时永辉[1] 罗阳[1,3] 王成[1,3] 陈熹[3] 曾科[3] 张辰宇[3] 夏正坤[2] 蔡晓懿[2] 汪俊军[1] 张春妮[1] 

机构地区：[1]南京军区南京总医院检验科,南京210002 [2]南京军区南京总医院小儿科,南京210002 [3]南京大学生命科学学院,南京210093

出　　处：《现代检验医学杂志》2013年第3期14-18,共5页Journal of Modern Laboratory Medicine

基　　金：国家自然科学基金资助项目（81171661;81270800）.

摘　　要：目的 检测分析原发性肾病综合征（NS）患儿血清全基因组微小核糖核酸（miRNA）表达谱,验证其中表达上调的miR-30a-5p,探讨其作为NS分子诊断指标的可能性.方法 收集南京军区南京总医院2010～2011年期间33例住院NS患儿未经治疗前血清和尿液样本,同时以30例年龄、性别匹配的健康儿童作对照.用高通量的TaqMan低密度芯片技术分别检测NS患儿和健康儿童的混合血清全基因组miRNA的表达量,再用TaqMan实时荧光定量PCR（qRT-PCR）方法对在患儿血清中表达上调的miR-30a-5p进行验证,同时测定miR-30a-5p在尿液中的含量.结果 低密度芯片结果显示,30种miRNA在患儿血清中的表达量是健康儿童的2倍以上,其中miR-30a-5p为正常对照的104倍.qRT-PCR检测证实,miR-30a-5p在患儿血清中的含量4.34±0.08,显著高于对照组1.00±0.02（Z=5.916,P〈0.000 1）,在患儿尿液中的水平也明显升高（3.44±0.05 vs 1.00±0.04）（Z=3.312,P=0.001）.用于诊断NS的血清miR-30a-5p ROC曲线下面积（AUCROC）为0.960（95% CI,0.915～1.000）,敏感度和特异度均为98.3%.相关性分析显示,血清或尿液中的miR-30a-5p含量与患儿年龄、性别、肾功能及血脂水平均无明显关联（P〉0.05）.结论 NS患儿血清miRNA表达谱与健康儿童有明显差异,miR-30a-5p在患儿血清及尿液中水平显著升高,与NS密切相关,是NS潜在的分子诊断指标.Objective To investigate microRNAs from a genome-wide serum microRNA expression profile of children with idiopathic nephrotic syndrome （NS） in order to identify differentially expressed miRNAs in NS and verify markedly up-regulated miR-30a-Sp and explore its potential as novel molecular marker for NS. Methods Serum samples were taken from 33 hospitalized NS children in Nanjing General Hospital from 2010 to 2011 year and 30 age-and gender-matched normal chil- dren. TaqMan Low Density Array was performed for screening the expression of miRNAs and then TaqMan qRT-PCR assay was conducted to confirm the upregulation of miR-30a-5p. The content of miR-30a-5p in urine was also been measured with qRT-PCR assay. Results TaqMan Low Density Array data showed that 30 miRNAs exhibited more than 2 fold expression level in NS than in normal control,and miR-30a-5p exhibited 104 fold. qRT-PCR confirmed that serum miR-30a-5p level was significantly elevated in NS as compared with normal controls （4. 34± 0. 08 vs 1.00 ± 0. 02） （Z = 5.916, P〈 0. 000 1）. The urinary miR-30a-5p content was also increased in NS （3.44±0.05 vs 1.00±0.04）（Z= 3. 312, P= 0. 001）. The AUC of serum miR-30a-5p was 0. 960 （95% CI 0. 915±1. 000） ,demonstrating a high sensitivity （98. 3 %） and specificity （98.3%） for NS prediction. Proteinuria and other kidney function and lipid parameters did not exhibit significant effects on serum or urinary miR-30a-5p concentration. Conclusion The expression profile of serum miRNAs was distinctive and miR-30a-5p content was significantly elevated in serum and urine samples from children NS,which holds potential as novel noninvasive bio- marker for NS.

关 键 词：原发性小儿肾病综合征 血清 尿液 微小核糖核酸 miR-30a-5p 

分 类 号：R692[医药卫生—泌尿科学] R392.2[医药卫生—外科学]