目的 本项研究采用高通量测序技术，筛选发展“快”“慢”慢阻肺［慢性阻塞性肺疾病（Chronic Obstructive Pulmonary Disease，COPD），简称慢阻肺］患者差异表达miRNA，验证其可能的生物学标志物及潜在治疗靶点。方法 收集新疆医科大学第四临床附属医院呼吸科2018年1月至2019年12月诊断明确慢阻肺稳定期患者240例，选取发展“快”组5例，发展“慢”组5例，正常组4例，健康对照组5例，健康人对照组5例。抽取受试者外周血，提取总DNA，进行miRNA高通量测序，运用生物信息学分析筛选不同发展速度慢阻肺患者各组间存在的差异性显著性miRNA、并进行GO和KEGG通路分析。候选的miRNA采用实时荧光定量PCR法在本地扩大样本的慢阻肺患者60例中进行验证。结果 与健康对照相比，COPD发展“快”患者中发现35个差异表达的miRNAs，COPD发展“慢”患者中发现16个差异表达的miRNAs，COPD发展正常患者中发现7个差异表达的miRNAs。根据差异miRNA生物信息学分析结合参考文献选择miR-4433a-5p、miR-1246、miR-1290进行qRT-PCR验证，结果显示miR-4433a-5p在COPD不同发展速度患者中显著升高，miR-1246、miR-1290在COPD不同发展速度患者中显著降低。结论 miR-4433a-5p、miR-1246、miR-1290可作为COPD疾病预测的标志物和治疗的潜在靶点。

Objective In this study, high-throughput sequencing technology was used to screen miRNA differentially expressed in patients with "fast" and "slow" COPD, and to verify its possible biological markers and potential therapeutic targets.Methods 240 patients with stable COPD diagnosed from January 2018 to December 2019 in the Department of Respiratory Diseases of the Fourth Clinical Affiliated Hospital of Xinjiang Medical University were collected. Five patients in the developing "fast" group and 5 patients in the developing "slow" group were selected. There were 4 cases in the group, 5 cases in the healthy control group, and 5 cases in the healthy control group. Peripheral blood was drawn from subjects, total DNA was extracted, miRNA high-throughput sequencing was performed, bioinformatics analysis was used to screen the significant differences in miRNAs between groups of COPD patients with different development speeds, and GO and KEGG pathway analysis were performed. Candidate miRNAs were verified by real-time fluorescent quantitative PCR in 60 cases of COPD patients whose samples were expanded locally.Results Compared with healthy control group, 35 differentially expressed miRNAs were found in patients with "fast" COPD development, 16 differentially expressed miRNAs were found in patients with "slow" COPD development, and 7 differentially expressed miRNAs were found in patients with normal COPD development. According to the analysis of differential miRNA bioinformatics combined with references, miR-4433a-5p, miR-1246, and miR-1290 were selected for qRT-PCR verification. The results showed that miR-4433a-5p was significantly increased in patients with different development speeds of COPD, and miR- 1246. miR-1290 is significantly reduced in COPD patients with different rates of development.Conclusion miR-4433a-5p, miR-1246, miR-1290 can be used as markers for predicting COPD and potential targets for treatment.

新疆维吾尔自治区自然科学基金青年项目（2019D01C177）；自治区卫生健康青年医学科技人才专项科研项目（WJWY-201957）