摘要
目的本项研究采用高通量测序技术,筛选发展“快”“慢”慢性阻塞性肺疾病(Chronic Obstructive Pulmonary Disease,COPD)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6例,发展“慢”组5例,正常组3例,健康对照组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.Six patients in the developing"fast"group and 5 patients in the developing"slow"group were selected.There were 3 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 realtime 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 7differentially 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.
作者
陶思冥
王益德
李风森
Tao Siming;Wang Yide;Li Fengsen(The fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830000,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2022年第9期3432-3441,共10页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
新疆维吾尔自治区卫生健康委员会青年医学科技人才专项科研项目(WJWY-201957):发展“快”“慢”COPD患者miRNA差异表达谱的构建及血清miRNA分子标志物的筛选与验证,负责人:陶思冥
新疆维吾尔自治区自然科学基金委员会青年项目(2019D01C177):基于TGF-β/Smad通路探讨益气固表丸含药学清对炎性肺上皮细胞免疫炎症机制的研究,负责人:陶思冥
关键词
慢阻肺
不同发展速度
MIRNA
高通量测序
Chronic obstructive pulmonary
Different developmental speed
miRNA
High-throughput sequencing technology
作者简介
通讯作者:李风森,本刊编委,博士,主任医师,主要研究方向:中西医结合呼吸病学研究。
