摘要
目的通过观察异体输血对创伤性失血患者外泌体miRNA的差异表达,筛选出差异表达的外泌体miRNA,为后续靶基因和通路的研究奠定基础。方法分别采集外伤患者在输异体血前和输血后外周血标本各3例,通过对血浆标本分离收获后的外泌体miRNA高通量测序,分析比较输血前后两组外泌体miRNA差异表达情况,同时以荧光定量PCR法进一步验证差异基因表达情况。结果通过高通量测序分析,共测序到1570个miRNA存在差异表达,实验组比较对照组,上调的有1189个,下调的有381个,以倍数变化Fold change≥2.0,P-value≤0.05作为差异miRNA筛选的阈值,筛选出显著变化的差异表达miRNA有14个,其中显著上调的miRNA有5个,分别为miR-499a-5p、miR-200a-3p、miR-372-3p、miR-novelchr12_5583、miR-novel-chr22_24253;显著下调的miRNA有9个,分别为miR-6852-5p、miR-452-5p、miR-1287-5p、miR-204-3p、miR-375-3p、miR-502-3p、miR-345-5p、miR-novel-chr7_38986、miR-451a。结论输注同种异体血液对创伤性失血患者的外泌体miRNA存在差异表达。其发生机制可能通过不同信号通路对患者机体免疫系统调节产生影响。然而,对患者机体免疫调节程度、与肿瘤的相关性有待于扩大样本量进一步研究。
Objective To observe the differential expression of exosomal miRNAs in patients with traumatic blood loss induced by allogeneic blood transfusion,screen out the differentially expressed exosomal miRNAs,and lay the foundation for the subsequent study of target genes and pathways.Methods Peripheral blood samples were collected from 3 trauma patients before and after transfusion,respectively.Through high-throughput sequencing of exosome miRNA extracted and harvested from plasma samples,the differential expression of exosome miRNA was analyzed and compared between the two groups.Meanwhile,fluorescence quantitative PCR was used to further verify the differential gene expression.Results High-throughput sequencing analysis found that a total of 1570 miRNA were differentially expressed.Compared with the control group,1189 miRNA were up-regulated and 381 miRNA were down-regulated in the experimental group.When fold change≥2.0,P-value≤0.05 were used as the threshold for differential miRNA screening,there were 14 differentially expressed miRNA with significant changes.5 miRNA were significantly up-regulated,including miR-499a-5p,miR-200a-3p,miR-372-3p,miR-novel-chr12_5583 and miR-novel-chr22_24253.9 miRNA were significantly down-regulated,including miR-6852-5p,miR-452-5p,miR-1287-5p,miR-204-3p,miR-375-3p,miR-502-3p,miR-345-5p,miR-novel-chr7_38986 and miR-451a.Conclusions Allogeneic blood transfusion triggers differential expression of exosomal miRNA in patients with traumatic blood loss.Its mechanism may affect the regulation of the patient's immune system through different signaling pathways.However,the degree of immune regulation in patients and the correlation with tumors need to be further studied with larger sample size.
作者
高宗帅
张海方
李志强
GAO Zongshuai;ZHANG Haifang;LI Zhiqiang(Shanghai Jiao Tong University Affiliated Sixth People's Hospital,Shanghai 200233)
出处
《临床输血与检验》
CAS
2023年第3期334-341,共8页
Journal of Clinical Transfusion and Laboratory Medicine
基金
上海市2020年度“科技创新行动计划”生物医药科技支撑专项项目(No.20S31902900)资助。
关键词
输血
外泌体
miRNA
高通量测序
Blood transfusion
Exosome miRNA
High-throughput sequencing
