膜性肾小球肾炎 非IgA系膜增生性肾小球肾炎患者全基因组miRNA表达谱的特征

彭武建+谭奎璧+黄建溶

[摘要]意图 了解非IgA系膜增生性肾小球肾炎（MsPGN）患者肾脏安排全基因组细小核糖核酸（miRNA）的表达特征及其效果。办法 别离搜集6例非IgA MSPGN患者肾穿安排和6例镜下病理查看正常的肾皮质安排作为MsPGN组和NRC组，运用Illumina高通量测序技能别离对其miRNA表达水平进行检测，剖析miRNA的表达差异，运用生物信息学技能开始剖析其功用。成果 在两组样本中具有显著性差异表达的miRNA有139种，其间在非IgA MSPGN组中表达上调的有13种，表达下调的有126种；别的还有20种miRNA在NRC组中特异性表达。生物信息学剖析显现这些miRNA靶基因与系膜增生有关。定论 非IgA MSPGN患者肾脏安排具有特定的miRNA表达，这些miRNA可能在疾病的发展中具有重要效果。

[关键词]细小RNA；非IgA系膜增生性肾小球肾炎；Illumina测序

[中图分类号] R692.3+1 [文献标识码] A [文章编号] 1674-4721（2016）06（c）-0004-04

[Abstract]Objective To investigate the expression characteristics and roles of the genome-wide microRNAs （miRNA） in non-IgA mesangial proliferative glomerulonephritis （MsPGN） kidney tissues.Methods Specimens from 6 patients with non-IgA MsPGN were collected by biopsy as MsPGN group，and 6 normal renal cortex specimens were collected as NRC group.The miRNA expression was evaluated by Illumina high-throughput sequencing technology.And the biological functions of special miRNA were predicted by the prediction software.Results In the MsPGN group，139 miRNA were found to be differentially expressed，13 miRNA were up-regulated and 126 miRNA were down-regulated.Moreover，20 miRNA were specifically expressed in NRC group.There was significant difference in miRNA expression between two groups.The bioinformatics showed that the regulated target genes of differentially expressed miRNA were associated with the mesangial proliferation.Conclusion The non-IgA MsPGN patients′ renal tissues have their own specific miRNA expression profiles and chromosome distribution characteristics.These miRNA may play an important role in the pathological changes of the kidney tissues.

[Key words]MicroRNA；Non-IgA Mesangial proliferative glomerulonephritis；Illumina sequencing

系膜增生性肾小球肾炎（mesangial proliferative glomerulonephritis，MsPGN）是我国原发性肾小球疾病中常见的病理类型，病理特征首要为系膜细胞及系膜外基质弥漫性增生，临床首要表现为蛋白尿和（或）血尿，30%的患者表现为肾病归纳征[1-3]。microRNA（miRNA）是一种内源性的非编码类小分子RNA，它在转录后具有调控基因的效果，人类体内有近1/3的基因受miRNA调控，并参加生命进程[4-5]。因而，本文通过高通量测序技能对MsPGN患者肾安排全基因组miRNA表达谱进行差异表达剖析，讨论MsPGN相关发病机制及寻觅新式MsPGN的医治靶点。

1材料与办法

1.1一般材料

挑选我院2014年6月～2015年6月接诊的12例 非IgA MsPGN患者，其间男5例，女7例；年纪28～63岁，均匀（43.68±9.12）岁；病程16～155 d。MsPGN组选取6例患者的肾皮质穿刺活检安排，样本通过临床、病理及实验室的查看进行归纳确诊，扫除其他继发性的要素，年纪（44.25±9.79）岁。NRC组选取6例行肾癌彻底治愈术患者的癌旁安排，其距肉眼癌团≥3 cm正常肾皮质安排，年纪（43.33±8.55）岁。两组的一般材料比较差异无统计学含义（P>0.05）。所用的标本由深圳市人民医院供给，并通过了医院道德委员会的赞同，患者知情且赞同。

1.2样本的处理和总RNA的提取[6]

将取出的样品在液氮中敏捷冷冻，冰箱温度坚持-80℃，直至送检。于常温下将样品冻结，依据Trizol试剂盒（Invitrogen）的说明书对总RNA进行提取。样品经剖析仪检测合格后（Agilent 2000 Bioanalyzer，合格规范RIN≥8.0且28S/18S>1.5），应用于小RNA（small RNA，sRNA）的建库与Illumina的测序。