基于RNA-Seq技术的亚低温防护大鼠放射性肺损伤的转录组学分析  

Transcriptome analysis of mild hypothermia protection against radiation-induced rat lung injury based on RNA-seq

作　　者：王远飞 王新钢[1] 许文黎[1] 王永丽[1] 黄立群[1] 岳娟[1] 安全[1] 李曙芳[1] WANG Yuanfei;WANG Xingang;XU Wenli;WANG Yongli;HUANG Liqun;YUE Juan;AN Quan;LI Shufang(Drug Safety Evaluation Center of China Institute for Radiation Protection,Shanxi Key Laboratory of Drug Toxicology and Drug for Radiation Injury,Key Laboratory on Radiotoxicology and Radiopharmaceutical Preclinical Evaluation,China National Nuclear Corporation,Taiyuan 030006,China)

机构地区：[1]中国辐射防护研究院药物安全性评价中心,药物毒理与放射损伤药物山西省重点实验室,中国核工业集团有限公司放射毒理与放射性药物临床前评价重点实验室,太原030006

出　　处：《中南大学学报（医学版）》2021年第4期345-350,共6页Journal of Central South University ：Medical Science

基　　金：山西省实验动物专项基金(2014k10)。

摘　　要：目的:分析放射性肺损伤大鼠的差异表达基因,从转录组水平揭示亚低温对大鼠放射性肺损伤的防护机制。方法:选取10只6~8周龄雄性SD大鼠并随机分为模型组与亚低温治疗组。建立放射性肺损伤大鼠模型,并对亚低温治疗组进行亚低温干预治疗,分别提取2组大鼠左侧肺组织RNA,用BGISEQ-500平台进行测序。采用edgeR差异分析软件分析2组基因表达的差异,对有统计学意义的差异表达基因进行基因本体(gene ontology,GO)功能富集分析和KEGG(Kyoto Encyclopedia of Genes and Genomes)通路富集分析,然后采用实时反转录聚合酶链反应(real-time reverse transcription PCR,real-time RT-PCR)对其中5个关键差异表达基因进行验证。结果:亚低温治疗组与模型组之间有2790个差异表达基因[错误发现率<0.001,|log_(2)(fold change)|>1],其中2257个基因上调,533个基因下调。5个关键基因real-time RT-PCR验证结果与转录组测序(RNA sequence,RNA-Seq)分析趋势一致。GO功能富集分析结果表明差异表达基因与细胞结合、代谢过程及细胞膜结构等有关;KEGG通路富集分析结果表明这些基因参与细胞黏附分子(cell adhesion molecules,CAMs)、雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)、紧密连接(tight junction)及核因子-κB(nuclear factor kappa-B,NF-κB)等重要生物学通路。结论:获得了亚低温防护大鼠放射性肺损伤相关差异表达基因及其显著富集通路,为亚低温技术防护放射性肺损伤提供了实验依据。Objective:To analyze the differentially expressed genes(DEGs)with radiation-induced rat lung injury,and to reveal the protective mechanism for mild hypothermia in the radiationinduced lung injury in rats at the transcriptome level.Methods:A total of 10 male SD rats aged 6-8 weeks were randomly divided into 2 groups to establish a rat model of radiation-induced lung injury,and one group was treated with mild hypothermia.RNA was extracted from left lung tissue of each group,and sequenced by BGISEQ-500 platform.Significance analysis of DEGs was carried out by edge R software.Gene ontology(GO)function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were used to analyze the gene function.Then 5 key DEGs were verified by real-time reverse transcription PCR(real-time RT-PCR).Results:There were 2790 DEGs(false discovery rate<0.001,|log_(2)(fold change)|>1)in the mild hypothermia group compared with the model group,in which 2257 genes were upregulated and 533 genes were down-regulated.When real-time RT-PCR was used to validate the 5 key genes,the result was consistent with the RNA-seq.GO functional enrichment analysis showed that these DEGs were related to cell binding,metabolic process and cell membrane structure,etc.KEGG pathway enrichment analysis showed that these genes were involved in important biological pathways such as cell adhesion molecules,mammalian target of rapamycin,tight junction,and NF-κB.Conclusion:The DEGs and pathways related to mild hypothermia protection against radiation-induced lung injury in rats are obtained,which provides an experimental basis for the protection of mild hypothermia against radiation-induced lung injury.

关 键 词：亚低温 紧密连接通路 RNA测序 放射性肺损伤 

分 类 号：R818[医药卫生—放射医学]