ISSN 1006-298X      CN 32-1425/R

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肾脏病与透析肾移植杂志 ›› 2014, Vol. 23 ›› Issue (4): 301-306.

• 论文 •    下一篇

局灶节段性肾小球硬化患者肾组织microRNAs表达分析

  

  • 出版日期:2014-08-28 发布日期:2014-09-02

Intrarenal miRNAs expression in patients with focal segmental glomerulosclerosis

  • Online:2014-08-28 Published:2014-09-02

摘要:

摘要 目的:比较局灶节段性肾小球硬化(FSGS)患者与正常对照肾组织中差异表达的microRNAs (miRNAs),探讨miRNAs在FSGS发病中的作用。 方法:选取年龄、性别匹配的FSGS患者6例及正常对照3例,留取肾组织标本,提取总RNA。将6个FSGS以及3个正常对照样品分别混合后,采用TaqMan低密度芯片(TaqMan Low Density Array)进行miRNAs表达谱分析,筛选出在FSGS患者肾组织中表达丰富并上调的miRNAs分子;然后选取14例FSGS患者和15例正常对照,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;采用Pearson相关分析FSGS患者肾组织中miRNAs表达水平与临床病理指标之间的相关性。 结果:芯片结果显示正常对照组检测到358个miRNAs,FSGS组检测到427个miRNAs。FSGS患者肾组织中有109个miRNAs表达上调。qRT-PCR结果显示与正常对照相比,FSGS患者肾组织中miRNA-155,miRNA-194和miRNA-215表达显著上调(P<0.05)。相关性分析显示,FSGS患者肾组织miRNA-215表达与肾小球硬化比例、总胆固醇水平正相关。 结论:FSGS患者肾组织miRNA-155,miRNA-194和miRNA-215表达上调,它们与FSGS发生和发展之间的联系值得进一步研究阐明。

关键词: 局灶节段性肾小球硬化, 肾组织, microRNAs

Abstract:

ABSTRACT Objective: To investigate the differences of miRNA expression between FSGS and normal kidney tissues. Methodology: Intrarenal miRNA expression was evaluated using TaqMan Low Density Array with pooled RNA samples from six FSGS patients and three normal controls. A set of differentially expressed miRNAs were found, and then validated by quantitative reverse transcription PCR (qRT-PCR) with individual samples. The relationships between miRNAs levels and clinical characters were analyzed by using Pearson correlation analysis. Results: The TaqMan Low Density Array experiment showed that 109 miRNAs were upregulated in the kidney of FSGS patients. After validation by qRT-PCR in individual samples, the intrarenal miRNA-155, miRNA-194 and miRNA-215 expressions were increased significantly in the patients with FSGS compared with normal controls. In addition, intrarenal miRNA-215 level was correlated significantly with serum total cholesterol levels and the proportions of sclerotic glomeruli in the patients. Conclusion: Intrarenal expression of miRNA-155, miRNA-194 and miRNA-215 in FSGS patients ws increased, which suggested these miRNAs may be involved in the pathogenesis of FSGS.
Key words

Key words: focal segmental glomerulosclerosis, intrarenal, microRNAs