海藻糖低温保存血小板miRNA差异表达分析

发布时间：2017-12-28 21:08

本文关键词：海藻糖低温保存血小板miRNA差异表达分析　出处：《第三军医大学学报》2016年18期 　论文类型：期刊论文

【摘要】：目的探讨海藻糖低温保存血小板过程中miRNA的变化。方法收集3份来自相同血型无偿献血员的单采血小板,少量混匀后,平均分为2组。对照组:新鲜血小板组(22℃,100%血浆储存的新鲜血小板2 h);实验组:海藻糖低温保存血小板组(10℃,70%海藻糖保存液储存5 d);采用高通量测序方法对两组样品进行了Small RNA测序,进行生物信息分析,主要包括已知miRNA鉴定、miRNA差异表达分析及富集分析。结果 2组样品经Small RNA高通量测序,平均每个样品获得了11兆净读值(Clean reads),经过生物信息分析,分别获得了821个已知miRNA。通过差异表达分析,在已知的miRNA中共有460个差异表达miRNA,其中194个miRNA表达上调,266个miRNA表达下调,其中显著差异表达已知miRNA共44个,分别在钙离子通路、MAPK通路、凋亡路径、轴突导向等储存损伤相关路径中富集程度较高。相对于对照组,实验组中hsa-miR-4449、hsa-miR-1296表达显著上调(P0.01,且差异倍数2),hsa-miR-665表达被抑制。结论血小板经过海藻糖低温保存后的miRNA呈现出特征性表达变化,提示miRNA活动参与了低温时海藻糖对血小板储存损伤的保护作用。
[Abstract]:Objective to investigate the changes of miRNA during the cryopreservation of trehalose at low temperature. Methods a total of 3 platelets from the same blood type unpaid blood donors were collected. After a small amount of mixing, the average was divided into 2 groups. Control group: fresh platelet group (22 DEG, 100% plasma stored fresh platelet 2 h); the experimental group: platelets group trehalose (10 DEG, 70% trehalose preservation liquid storage for 5 d); using high-throughput sequencing method in two groups of samples were Small RNA sequencing, analysis of information, including analysis and enrichment analysis of miRNA identification, the difference of miRNA expression is known. Results 2 groups of samples were sequenced by Small RNA high throughput sequencing. The average sample obtained 11 trillion net reading value (Clean reads). After bioinformatics analysis, 821 known miRNA were obtained. Through the analysis of the differences in expression, known as miRNA, a total of 460 differentially expressed miRNA, of which 194 up-regulated the expression of miRNA and 266 miRNA expression, the significant difference of the expression of known miRNA a total of 44, the damage degree of enrichment of the relevant path in high calcium ion pathway, MAPK pathway, apoptosis pathway, axon guidance and storage respectively. Compared with the control group, the expression of hsa-miR-4449 and hsa-miR-1296 in the experimental group was significantly up (P0.01, and the difference multiplier was 2), and the expression of hsa-miR-665 was inhibited. Conclusion the miRNA expression of platelets after trehalose cryopreservation showed a characteristic expression change, suggesting that miRNA activity is involved in the protective effect of trehalose on platelet storage damage at low temperature.
【作者单位】：第三军医大学西南医院输血科中国人民解放军重庆血站;
【基金】：国家自然科学基金面上项目(81270649)~~
【分类号】：R446.1
【正文快照】： 血小板在保存过程中结构和功能会随着时间延长而发生变化,这些改变统称为储存损伤(platelet storagelesion,PSL)[1]。常规的血小板保存条件为22℃振荡保存5 d。为了减少血小板的储存损伤,人们尝试采用低温保存方法,低温保存虽可以延长血小板保存时限,但低温易致血小板发生活化

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