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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 screen and identify specific miRNAs associated with Mycoplasma pneumoniae pneumonia. Methods: The blood of children with Mycoplasma pneumoniae pneumonia was collected. The changes of miRNAs expression in blood lymphocytes of children with Mycoplasma pneumoniae pneumonia and normal persons were analyzed by miRNA microarray. The accuracy of the results of microarray detection was verified by RT-PCR. Mycoplasma pneumonia-associated specific miRNAs. Using target gene software to predict each possible target gene, mRNA expression difference was verified by RT-PCR. Results: There were 105 miRNAs that were upregulated in the blood lymphocytes of children with Mycoplasma pneumoniae pneumonia, down-regulated by 133. Among them, 30 were up-regulated (more than 5-fold), 10 up-regulated (more than 10-fold), and 133 down-regulated (less than 0.5-fold) (P <0.05). The expression of miR-20a-3p, miR-100-5p, miR-93-5p and let-7b-5 were detected by RT-PCR. , TGFβR1 mRNA expression decreased to varying degrees. Conclusion: miRNAs are differentially expressed in blood lymphocytes of children with Mycoplasma pneumoniae pneumonia, and these differentially expressed miRNAs and their potential target genes may be involved in the infection and immune response caused by Mycoplasma pneumoniae.