【摘 要】
：
目的 了解在体外常规贮存血小板微小RNA(miRNA)的表达谱变化,探讨miRNA参与调控血小板贮存损伤(PSL)的分子机制.方法 留取来自无偿献血者的单采血小板20(人)份(5 mL/份),摇匀后等量混合,贮存于(22±2)℃恒温血小板振荡保存箱中,分别在d1和d5取样,采用纳米球(DNB)测序技术对血小板miRNA组测序;2组(不同贮存期)血小板相比miRNA表达量差异≥2倍(P＜0.01)时,采用miRanda和TargetScan软件预测靶基因及基因本体论(GO)功能富集和京都基因与基因组百科全书
【机 构】
：
山东省血液中心,山东济南250014;菏泽市中心血站
论文部分内容阅读
目的 了解在体外常规贮存血小板微小RNA(miRNA)的表达谱变化,探讨miRNA参与调控血小板贮存损伤(PSL)的分子机制.方法 留取来自无偿献血者的单采血小板20(人)份(5 mL/份),摇匀后等量混合,贮存于(22±2)℃恒温血小板振荡保存箱中,分别在d1和d5取样,采用纳米球(DNB)测序技术对血小板miRNA组测序;2组(不同贮存期)血小板相比miRNA表达量差异≥2倍(P＜0.01)时,采用miRanda和TargetScan软件预测靶基因及基因本体论(GO)功能富集和京都基因与基因组百科全书(KEGG)信号通路富集分析.采用实时荧光定量PCR(qPCR)方法检测miRNA的表达以验证DNB测序结果.结果 miRNA表达谱:贮存d5与d1血小板相比,共有315个表达量差异明显的miRNA,其中上调146个(如miR-146a、let-7b等),下调169个(如miR-30d、miR-142等);已知126个miRNA中,表达上调的43个,表达下调83个;新见miRNA序列189条.d5与d1组差异表达miRNA靶基因呈明显富集的GO条目(term)包括细胞组分、细胞器、细胞膜等细胞结构,黏附、催化、活性活性等分子功能,细胞加工、代谢、生物调节、应激等生物过程.相应的KEGG富集前10的通路中主要是信号转导、分泌、膜转运、氨基酸代谢、多糖代谢、蛋白质合成、环境适应等功能.随机挑选的6个差异表达miRNA所做qPCR结果与测序结果均一致.结论 随着血小板体外贮存时间的延长,miRNA表达谱发生明显变化;功能预测提示这些miRNA可能参与体外贮存下血小板发生PSL的调控.
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