ARDS 患者中血清 microRNAs 生物标志物的筛选

摘要

Objective To screen serum miRNAs biomarker in patients with acute respiratory distress syndrome(ARDS).Methods 60 patients with ARDS in the intensive care unit of our hospital from June 2013 to June 201 5 were collected,and 50 volunteers in our hospital from June 2013 to June 201 5 were collected as a control group.There was not statistical significance in age and gender between the two groups.The serum miRNAs expression profiling in patients with ARDS was made by microarray analysis. The microarray data were further analyzed by real-time quantitative RT-PCR.Results A260/A280 and A260/A230 ratio of extracted total RNA in this experiment were greater than 1.9 and 2.0,respectively, indicating that the quality of the extracted total RNA was preferred.The miRNA microarray data showed that 27 species miRNAs expression was significantly altered compared with the healthy control group based on SAM analysis (P < 0.05),of which 20 kinds of miRNAs expression was upregulated,seven miRNAs expression was downregulated.The downregulated miRNAs included miR-24,miR-26a,miR-126,and family members of Let-7,the upregulated miRNAs included miR-99a,miR-127,miR-128b,miR-135b,miR-30a,and miR-30b.The final miRNAs singled out were validated through RT-PCR,miR-99a and miR-30b were confirmed to be upregulated in patients with ARDS,and miR-126 and miR-26a were confirmed to be downregulated in patients with ARDS.Conclusions The combination of miR-99a,miR-30b,miR-126 and miR-26a may be a potential diagnostic biomarker of ARDS.%目的：对 ARDS 患者血清 miRNAs 生物标志物进行筛选。方法选取我院 ICU 2013年6月至2015年6月接收的60例 ARDS 患者,同时选取我院体检的志愿者50名作为健康对照组,2组患者的年龄、性别差异无统计学意义。ARDS 患者血清 miRNAs 的表达图谱通过微阵列分析。微阵列数据通过实时定量 RT-PCR 进一步分析。结果本实验所提取的总 RNA 的 A260/A280和 A260/A230比例分别大于1.9和2.0,表明所提取的总 RNA 的质量是较好的。miRNAs 微阵列数据的结果表明：基于 SAM 分析有27种 miRNAs 的表达与健康对照组相比是显著性改变的(P <0.05),其中有20种 miRNAs 表达是上调的,有7种 miRNAs 表达是下调的。下调的 miRNAs 包括 miR-24、miR-26a、miR-126和 Let-7的家庭成员,上调的 miRNAs 包括 miR-99a、miR-127、miR-128b、miR-135b、miR-30a 和 miR-30b。最终挑选出来的 miRNAs 是经过 RT-PCR 验证的,miR-99a 和 miR-30b 被确认是 ARDS 患者血清中上调的 miRNAs,而 miR-126和 miR-26a 被确认是 ARDS 患者血清中下调的miRNAs。结论 miR-99a、miR-30b、miR-126和 miR-26a 这4个血清组合可能作为 ARDS 诊断潜在的生物标志物。