肺炎支原体肺炎患儿血液中差异表达miRNAs的筛选及鉴定

发布时间：2018-07-17 03:01

【摘要】：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(P0.05)。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。
[Abstract]:Objective: to screen and identify the specific miRNAs. method related to Mycoplasma pneumoniae pneumonia: to collect the blood of children with Mycoplasma pneumoniae pneumonia and to analyze the changes in the expression of miRNAs in the blood lymphocytes of children with Mycoplasma pneumoniae pneumonia and normal people by miRNA chip technology, and verify the accuracy of the results by the RT-PCR method and confirm the results. The specific miRNAs. related to Mycoplasma pneumoniae pneumonia was used to predict the possible target genes by target gene software, and the difference of mRNA expression was verified by RT-PCT. Results: the blood lymphocytes of children with Mycoplasma pneumoniae were screened out of up regulated miRNA 105, 133 down-regulation. The up regulation (more than 5 times) was up to up (greater than 10 times). 10, down down (less than 0.5 times) of 133 (P0.05). MiR-20a-3p, miR-100-5p, miR-93-5p, and let-7b-5 were selected for subsequent RT-PCR verification, and the results were consistent with the chip. Bcl-2, IGF-1R, PTEN, TGF beta R1 mRNA expression was reduced in varying degrees. Conclusion: there is a blood lymphocyte in children with Mycoplasma pneumoniae pneumonia. Differential expression of these differentially expressed miRNAs and its potential target genes may be involved in the infection and immune response caused by Mycoplasma pneumoniae.
【作者单位】：南京医科大学附属儿童医院呼吸科;马鞍山市十七冶医院儿科;
【基金】：国家自然科学基金(青年科学基金项目)(81200012)
【分类号】：R725.6

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