肺炎支原体肺炎患儿血液中差异表达miRNAs的筛选及鉴定

Identification of differentially expressed bloody miRNAs in children with Mycoplasma pneumoniae pneumonia

目的 :筛选鉴定与肺炎支原体肺炎相关的特异性miRNAs。方法 :收集肺炎支原体肺炎患儿的血液,通过miRNA芯片技术分析肺炎支原体肺炎患儿和正常人血液淋巴细胞中miRNAs的表达谱变化,通过RT-PCR方法验证芯片检测结果的准确性,确认与肺炎支原体肺炎相关的特异性miRNAs。利用靶基因软件预测各自可能的靶基因,利用RT-PCT验证mRNA表达差异。结果:肺炎支原体肺炎患儿血液淋巴细胞中筛选出表达上调的miRNA 105个,表达下调的133个。其中上调(大于5倍)的30个,上调(大于10倍)的10个,下调(小于0.5倍)的133个(P<0.05)。选取表达差异显著的miR-20a-3p、miR-100-5p、miR-93-5p以及let-7b-5进行后续RT-PCR的验证,结果与芯片一致;Bcl-2、IGF-1R、PTEN、TGFβR1 mRNA表达有不同程度的降低。结论:miRNAs在肺炎支原体肺炎患儿血液淋巴细胞中存在差异表达,而这些差异表达的miRNAs及其潜在靶基因可能参与肺炎支原体导致的感染与免疫反应。

Objective：To explore related miRNAs to Mycoplasmapneumoniae（Mp） caused respiratory infections and immune reaction. Methods： Blood of children with Mp pneumonia （MPP） was collected. By miRNA microarray technology, miRNAs expression differences in lymphocytes were analysed between MPP and control group. RT-PCR was used to verify the accuracy of microarray results. Specific miRNAs associated with MPP were explored. Target gene software was used to predict the possible target genes, and RT-PCT was used to verify the mRNA expression difference. Results：Compared with normal group,in MPP group 105 miRNAs increased（with 30 more than 5 fold change, 10 more than 10） and 133 decreased （with 133 less than 0.5 fold） （P〈0.05）. Significantly differentially expressed miRNAs ：miR-20a-3p ,miR-100-5p ,miR-93-5p, and let-7b-5p increased detected by RT-PCR, and were consistent with the array results. As potential target genes, Bcl-2, IGF-1R ,PTEN, and TGFI3R1 mRNA expression decreased. Conclusion：miRNAs differential expressed in blood lymphocytes in children with MPP,and the difference may be involved in infection and the immune response caused by Mp.