基于RNA-Seq技术大鼠放射性肺损伤转录组学分析

Transcriptomic analysis of radiation-induced lung injury in rats based on RNA-Seq technology

目的分析放射性肺损伤(RILI)大鼠的差异表达基因,从转录组水平探讨大鼠RILI的机制。方法选取40只SD大鼠并随机分为实验组(30 Gy右肺照射组)与对照组。建立RILI大鼠模型,提取两组大鼠右肺组织RNA,采用Illumina平台进行测序。采用DESeq2软件(1.20.0)分析两组的基因表达变化,并采用实时逆转录聚合酶链反应对其中6个关键差异表达基因进行验证。对差异表达基因进行聚类分析、GO功能富集分析及KEGG通路富集分析。结果转录组学分析实验组与对照组之间有414个差异表达基因[P<0.05,|log2(FC)|>1.0],其中,270个上调基因,144个下调基因。根据基因的差异倍数和所富集到的炎症相关的通路筛选出的关键差异基因包括Fos、Cxcl2、Nr1d1、Angptl4、Ccn4、Tnc,实时逆转录聚合酶链反应验证结果与转录组测序分析趋势一致。GO功能富集分析的结果显示,差异基因主要参与染色体分离、核分裂、细胞器裂变等过程;KEGG通路富集分析的结果显示,差异基因参与昼夜节律、ECM受体相互作用、cAMP信号通路、TNF信号通路等重要生物学过程。结论本研究初步分析获得了大鼠RILI相关差异表达基因及其相关通路,为RILI的临床诊治提供了实验依据。

Objective To analyze the differentially expressed genes in rats with radiation-induced lung injury(RILI)and explore the mechanism of RILI from the transcriptome level.Methods A total of 40 SD rats were randomly selected and divided into experimental group(30 Gy right lung radiation in rats)and control group.The rat model of RILI was established.The RNA of the right lung tissue of the two groups was extracted and sequenced by Illumina platform.The differences of gene expression between the two groups was analyzed by DESeq2(1.20.0).Then the six of these key differentially expressed genes were verified by real-time reverse transcription polymerase chain reaction.The differentially expressed genes were subjected to cluster analysis,GO function enrichment analysis and KEGG pathway enrichment analysis.Results Transcriptomics analysis showed that there were 414 differentially expressed genes between the experimental group and the control group[P<0.05,|log2(FC)|>1.0],including 270 up-regulated genes and 144 downregulated genes.According to the differential fold of genes and the inflammation related pathway enriched by differential genes,the key differential genes selected include Fos,Cxcl2,Nr1d1,Angptl4,Ccn4,Tnc,they were confirmed by real-time reverse transcription polymerase chain reaction,and the differential gene expression change trends were consistent with the RNA sequence analysis conclusions.The results of GO function enrichment analysis showed that the differential genes were mainly involved in chromosome separation,nuclear division,organelle fission processes.The results of KEGG pathway enrichment analysis showed that the differential genes were involved in circadian rhythm,ECM receptor interaction,cAMP signaling pathway and TNF signaling pathway.Conclusion This study preliminarily analyzed the differentially expressed genes and related pathways related to RILI in rats,and provided experimental basis for prevention and treatment of RILI.