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目的借助微小RNA(microRNA,miRNA)芯片研究IgA肾病(IgA nephropathy,IgAN)中miRNA的差异表达。方法收集11例IgAN患者肾穿标本作为IgAN组,3例镜下病理检查正常的肾皮质作为对照组。使用Trizol试剂提取每组总RNA,分离miRNA,采用miRNA芯片筛选出差异表达显著的miRNA,荧光实时定量PCR(Real-timePCR)技术验证芯片结果的可靠性。结果IgAN组中miRNA表达出现显著差异的有66个,其中表达显著上调35个,显著下调31个。挑选hsa-miR-637、hsa-miR-492进行Real-timePCR相对定量检测,结果显示2个miRNA的表达趋势与芯片结果相符,证明了芯片结果的可靠性。结论IgAN是一种经典的多因素、多基因病,本实验通过芯片技术表明miRNA在IgAN中出现显著差异表达。
Objective To study the differential expression of miRNAs in IgA nephropathy (IgAN) by means of microRNA (miRNA) microarray. Methods The renal peritoneum specimens from 11 patients with IgAN were collected as IgAN group and 3 patients with pathologically normal kidney cortex as control group. Trizol reagent was used to extract total RNA from each group, miRNAs were isolated, miRNAs with significant differential expression were screened by miRNA chip, and the reliability of chip results was verified by real-time quantitative PCR (Real-time PCR). Results There were 66 miRNAs in IgAN group, of which 35 were significantly up-regulated and 31 were significantly down-regulated. The relative quantification of hsa-miR-637 and hsa-miR-492 was carried out by Real-time PCR. The results showed that the expression trend of the two miRNAs was consistent with the chip results, which proved the reliability of the chip results. Conclusion IgAN is a classic multifactorial and polygenic disease. In our experiment, miRNAs were significantly differentially expressed in IgAN by microarray.