机采血小板储存过程中5种miRNA前体的表达变化  

Detection of 5 pre-miRNAs in human apheresis platelets during storage by qRT-PCR

作　　者：叶银才[1] 李强[2] 蔡雪娇[1] 郭瑞德[1] 俞石芳[3] 

机构地区：[1]温州医科大学附属第一医院输血科,浙江温州325000 [2]温州医科大学附属第三医院 [3]浙江大学医学院附属第二医院

出　　处：《中国输血杂志》2015年第10期1222-1225,共4页Chinese Journal of Blood Transfusion

基　　金：浙江省自然科学基金青年基金(Q12H200001);温州市科技局社会发展项目(Y20110051)

摘　　要：目的分析机采血小板储存过程中表达量有明显升高的5个miRNA前体(pre-miRNA)表达谱,为研究机采血小板中miRNA的成熟及调节机制提供参考。方法分别提取不同储存时间(1、3、5 d)各2 m L机采血小板总RNA,以5S rRNA为内参照,采用实时荧光定量(qRT-PCR)法分别检测5个miRNA前体pre-miR-16、-96、-150、-155和-316的表达水平。结果以新鲜机采血小板0 d pre-miRNA表达水平为基准,pre-miR-16的相对表达量在1、3、5 d分别为0.98±0.08、0.96±0.11、0.92±0.14;pre-miR-96的相对表达量在1、3、5 d分别为0.97±0.08、0.98±0.09、0.94±0.12;pre-miR-150的相对表达量在1、3、5 d分别为0.95±0.09、0.91±0.06、0.93±0.15;pre-miR-155的相对表达量在1、3、5 d分别为0.97±0.12、0.95±0.08、0.95±0.10;pre-miR-326的相对表达量在1、3、5 d分别为0.98±0.08、0.96±0.12、0.86±0.18。相对于新鲜血小板,上述5种pre-miRNA在储存过程中表达量无明显改变(P>0.05)。结论机采血小板储存过程中表达量明显升高的5个miRNA的pre-miRNA表达相对稳定,并未伴随miRNA表达改变而改变。Objective To analyze the differential expression of their precursor （pre-miRNAs） in apheresis platelets during storage, and to explore the relationship between pre-miRNAs and miRNAs during storage. Methods Based on the miRBase, the primers and the fluorogenic probe were designed and synthesized. The quantitative real-time PCR （qRT-PCR） were used to detect the relative expression levels of pre-miRNAs in apheresis platelets at different preservation time （ 1,3 and 5 day）. Fresh apheresis platelets were used as Day 0 platelets. Results As compared to the fresh apheresis platelets, the relative expression levels of pre-miR-16 at different preservation time （ 1,3 and 5 day ） were 0. 98 ± 0. 08, 0.96 ± 0. 11 and 0. 92 ±0. 14, respectively; the levels of pre-miR-96 were 0. 97 ± 0. 08, 0.98 ± O. 09 and, 0. 94 ± 0. 12, respectively; the levels of pre-miR-150 were 0.95 ±0. 09, 0. 91 ±0.06 and 0. 93 ±0. 15, respectively; the levels of pre-miR-155 were 0. 97 ± 0.12, 0.95 ± 0. 08 and 0. 95 ± 0. 10, respectively ; and the levels of pre-miR-326 were 0. 98 ± 0.08, 0. 96 ± 0.12 and 0. 86 ± 0. 18, respectively. The observation inferred that the relative expression levels of the 5 pre-miRNA had no significant changes at different preservation time of storage. Compared to fresh apheresis platelets, the expression levels of the 5 pre- miRNAs had not changed accompanied by their miRNA ssignificantly during storage. Conclusion In platelets, the matura- tione of miRNAs beginsan with the unspliced pre-miRNAs, which are mostly originated from the cytoplasm of macrophages. Other regulatory network of miRNA is speculated to existed in platelets during storage.

关 键 词：血小板 MIRNA miRNA前体 实时荧光定量PCR 血小板储存损伤 

分 类 号：R446.1[医药卫生—诊断学] R331.143[医药卫生—临床医学]