微小RNA作为单采血小板储存损伤标志物不同保存期的差异表达谱

Differential expression profiles of microRNA as markers of apheresis platelet storage lesion in different storage periods

目的:探讨单采血小板不同保存期微小RNA(mRNA)表达谱差异,发现由于储存过程中血小板生物学变化引起的新的敏感性表达miRNA。方法:通过新一代small RNA测序技术结合生物信息学分析方法,筛选出单采血小板在体外保存期间存在差异表达的miRNA,并采用实时荧光定量PCR(qRT-PCR)法验证。结果:miRNA表达谱芯片结果显示6组不同保存期单采血小板普遍高表达hsa-let-7家族,hsa-miR-26a-5p, hsa-miR-92a-3p, hsa-miR-199,hsa-miR-103a-3p。时间序列分析对944个差异表达miRNA进行趋势分类,共发现44个基因呈现14种趋势变化。以保存第1天为对照组,hsa-miR-4433b-5p, hsa-miR-22-3p, hsa-miR-30c-5p显著下降。结论:分析筛选出表达差异的miRNA-30c-5p, miRNA-22-3p, miRNA-4433b-5p为下一步研究单采血小板储存损伤标志物奠定基础。

Objective: To discuss the difference of microRNA expression profile in different storage periods of apheresis platelets and identify a new collection of miRNAs that showed a sensitivity expression pattern miRNA-30, miRNA-22, miRNA-4433 due to biological platelet changes during storage. Methods: Through a new generation of small RNA sequencing technology and bioinformatics analysis methods, the microRNAs that were differentially expressed during the storage of apheresis platelets in vitro were screened. The expression of miRNA was verified by real-time fluorescence quantitative PCR(qRT-PCR). Results: The miRNA expression profile results showed that the 6 groups of apheresis platelets with different storage periods generally highly expressed hsa-let-7 family, hsa-miR-26a-5p, hsa-miR-92a-3p, hsa-miR-199 and hsa-miR-103a-3p. Time series analysis performed trend classification of 944 differentially expressed miRNAs, and found that 44 genes showed 14 trend changes. Taking the first day of storage as the control group, hsa-miR-4433b-5p, hsa-miR-22-3p, and hsa-miR-30c-5p were significantly down-regulated. Conclusion: The analysis of differentially expressed miRNA-30c-5p, miRNA-22-3p, miRNA-4433b-5p may lay the foundation for further research on apheresis platelet storage lesion.