核黄素光化学处理与未处理后不同贮存时间血小板miRNA表达谱的差异分析  

Comparative analysis of miRNA profiles of platelets treated and untreated with riboflavin and ultraviolet-B light

作　　者：刘群 庄云龙[1] 王玉霞[1] 叶辉[1] 谯铭铭 盖厦 陈元锋[1] 申华[1] 蒋保云 LIU Qun;ZHUANG Yunlong;WANG Yuxia;YE Hui;JIAO Mingming;GAI Xia;CHEN Yuanfeng;SHEN Hua;JIANG Baoyun(Blood Center of Shandong Province,Jinan 250014,China)

机构地区：[1]山东省血液中心,山东济南250014

出　　处：《中国输血杂志》2021年第7期701-707,共7页Chinese Journal of Blood Transfusion

基　　金：山东省自然科学基金面上项目(ZR2017MH070);山东省医药卫生科技发展计划项目(2018WSB09004);中国输血协会威高科研基金资助项目(CSBT-WG-2017-03);山东省血液中心项目(201710)。

摘　　要：目的分析核黄素光化学技术(VB_(2)-PRT)处理与否的贮存d1和d5血小板微小RNA(miRNA)表达谱的变化,探讨VB_(2)-PRT处理miRNA参与调控血小板发生贮存损伤(PSL)的分子机制。方法留取无偿献血者单采血小板20(人)份(5 mL/份),混合摇匀后均分为2等份,1份为VB_(2)-PRT处理组(E组):使用终浓度为50μmol/L VB_(2)和6.24 J/mL紫外线(UV)B光处理8 min;另1份为对照组(C组):不做处理;2组皆贮存于(22±2)℃恒温血小板振荡保存箱中。在贮存d1和d5分别取样(5 mL),分别命名为E1、E5、C1和C5组,采用纳米球(DNB)测序技术对其做血小板miRNA组测序,采用DEGseq和MA-plot分析软件筛选E和C组间差异表达的小RNA,当E和C组间的血小板miRNAs表达差异≥2倍(P<0.01)时,对这部分血小板miRNA做靶基因预测及GO功能富集和KEGG信号通路富集分析。结果E1和C1组血小板miRNA的表达谱相比:在E1组中筛选出487个表达差异明显(P<0.01)的miRNAs,包括220个miRNAs表达上调,如miR-146a和let-7b等,267个表达下调,如miR-7和miR-1260等;E5和C5组血小板miRNA的表达谱相比:在E5中筛选出229个表达差异明显(P<0.01)的miRNAs,包括80个miRNAs表达上调,如miR-423和miR-378等,149个表达下调,如miR-451和miR-30等。E1与C1组以及E5与C5组的差异表达miRNAs靶基因呈明显富集的GO条目(term)数相似,包括细胞组分、细胞器、细胞膜等细胞结构,黏附、催化、分子转换、运输、转录因子和受体活性等分子功能,细胞加工、代谢、生物调节、应激等生物过程。E5与C5组相应的KEGG富集前10的通路中,较E1与C1组不同的是缺少涉及环境适应、翻译和黏蛋白合成信号通路,却增加了肌醇磷酸代谢、磷脂酰肌醇信号系统和趋化因子信号通路。结论经VB_(2)-PRT处理的血小板贮存一段时间其miRNAs表达谱发生明显变化;功能预测提示这些miRNA可能参与VB_(2)-PRT处理下血小板发生PSL的调控。Objective To analyze the changes of microRNA(miRNA)expression profiles on day 1 and day 5 after storage with or without riboflavin and ultraviolet-B(UVB)light(VB_(2)-PRT)treatment,and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion(PSL)under VB_(2)-PRT treatment.Methods 20 apheresis platelet concentrates(5 mL/sample),collected from voluntary donors,were split into two group after mixing and agitation.One was treated with riboflavin(final concentration 50μmol/L)plus 6.24 J/mL UVB light(E group),and the other worked as a control group(C group)without any treatment.Both groups were subjected to agitated storage at(22±2)℃horizontally.The platelet concentrates were sampled on d1 and d5(5 mL)during storage,named as E1,E5,C1 and C5 groups,respectively,and sequenced by DNA nanoball(DNB)sequencing technology.The differentially expressed miRNAs between E and C groups were screened by using DEGseq and MA-plot analysis software,and GO function enrichment analysis and KEGG pathway enrichment analysis were further performed when the different expression between groups reached twofold and above.Results Compared with C1 group,487 miRNAs with significantly different expression(P<0.01)were screened in E1 group,including 220 up-regulated miRNAs,such as miR-146 a and let-7 b,and 267 down regulated miRNAs,such as miR-7 and miR-1260.Compared with C5 group,229 miRNAs with significantly different expression(P<0.01)were screened in E5,including 80 miRNAs with up-regulated expression,such as miR-423 and miR-378,and 149 down regulated miRNAs,such as miR-451 and miR-30.The target genes with differentially expressed miRNAs in E1 vs C1 groups and E5 vs C5 groups were similar in the numbers of enriched GO terms,including cell components,organelles,cell membrane and other cell structures,molecular functions such as adhesion,catalysis,molecular transformation,transportation,transcription factors and receptor activity,cell processing,metabolism,biological regulation,stress and other biologica

关 键 词：miRNA表达谱 血小板 核黄素光化学技术 血小板贮存损伤 靶基因功能分析 

分 类 号：R457.12[医药卫生—治疗学] R331.143[医药卫生—临床医学]