摘要
目的筛选和验证脓毒症患儿外周血中差异表达的长链非编码RNA(long non-coding RNA,lncRNAs),探讨其在儿童脓毒症发病机制中的作用。方法选取2016年1月至12月在首都儿科研究所附属儿童医院ICU进行救治的3例脓毒症患儿和3例健康儿童的外周血标本,通过lncRNA测序技术筛选出差异表达的lncRNAs和mRNAs;对差异表达的lncRNAs进行靶基因预测,并根据GO分析结果选出与线粒体ATP合酶(F1FO-ATPase)活性相关的lncRNA-mRNA关系对;进一步扩大样本量,采用实时荧光定量逆转录-聚合酶链反应(qRT-PCR)技术对筛选结果中F1FO-ATPase活性相关的部分mRNAs及lncRNAs进行验证。结果测序结果显示,与健康儿童相比,脓毒症患儿外周血中差异表达的lncRNAs共252个(86个表达上调,166个表达下调),差异表达的mRNAs共2652个(955个表达上调,1697个表达下调);qRT-PCR验证结果显示:lncRNA ENST00000621933.1、ENST00000616950.1、ENST00000595748.1在脓毒症患儿中表达升高(P<0.05),MT-ATP8、ATP5E以及lncRNA ENST00000624705.1和ENST00000615535.1在脓毒症患儿中表达降低(P<0.05),与测序结果一致。结论脓毒症患儿外周血中存在差异表达的lncRNAs,以MT-ATP8及ATP5E作为靶基因的lncRNA ENST00000621933.1、ENST00000616950.1、ENST00000595748.1、ENST00000624705.1和ENST00000615535.1的表达水平可能与儿童脓毒症的发生发展有关。
Objective To screen and identify differentially expressed long non-coding RNA(lncRNAs)in peripheral blood of children with sepsis,and to explore the role of lncRNAs in the pathogenesis of sepsis in children.Methods The peripheral blood samples of 3 children with sepsis admitted to the ICU of Children′s Hospital of Capital Institute of Pediatrics from January to December 2016 and 3 healthy children who underwent physical examination in our hospital during the same period were selected,and the differential expression profiles of lncRNAs and mRNAs were screened by lncRNAs sequencing technology.The target genes of differentially expressed lncRNAs were predicted and the relationship pairs of lncRNA-mRNA related to F1FO-ATPase activity were constructed according to the results of GO analysis.Further increasing the sample size,we verified the expression of some F1FO-ATPase activity-related mRNAs and lncRNAs which were differentially expressed in the screening results by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR).Results Sequencing results showed that there were 252 lncRNAs with significant differential expression in peripheral blood of children with sepsis compared with healthy children,of which 86 were up-regulated and 166 were down-regulated;meanwhile,there were 2652 mRNAs with significant differential expression,of which 955 were up-regulated and 1697 were down-regulated.The results of qRT-PCR showed that the expression of lncRNA ENST00000621933.1,ENST00000616950.1 and ENST00000595748.1 in peripheral blood of children with sepsis increased(P<0.05),while the expression of MT-ATP8,ATP5E and ENST00000624705.1,ENST00000615535.1 in peripheral blood of children with sepsis decreased(P<0.05),which was consistent with the sequencing results.Conclusion lncRNAs are differentially expressed in peripheral blood of children with sepsis compared with healthy children.The expression levels of lncRNA ENST00000621933.1,ENST00000616950.1,ENST00000595748.1,ENST00000624705.1 and ENST00000615535.1 which their
作者
王圆圆
卢秀秀
陈源美
李宁
李伟
孙中媛
郭琳瑛
崔小岱
宋国维
张琪
Wang Yuanyuan;Lu Xiuxiu;Chen Yuanmei;Li Ning;Li Wei;Sun Zhongyuan;Guo Linying;Cui Xiaodai;Song Guowei;Zhang Qi(Department of Critical Care Medicine,Capital Institute of Pediatrics-Peking University Teaching Hospital,Beijing 100020,China;Department of Pediatrics,Peking University China-Japan Friendship School of Clinical Medicine,Beijing 100029,China;Central Laboratory,Capital Institute of Pediatrics,Beijing 100020,China)
出处
《中国小儿急救医学》
CAS
2020年第4期272-278,共7页
Chinese Pediatric Emergency Medicine
基金
北京市医院管理局临床技术创新项目(XM201412)
北京市自然科学基金(7172037)。
