APAP急性肝损伤小鼠肝脏组织转录组RNA-seq及相关信号通路分析

RNA-seq Analysis Reveals Novel Genes and Signaling Pathways Associated with APAP-induced Acute Liver Injury in Mice

[目的]筛选与对乙酰氨基酚(acetaminophen,APAP)诱导的急性肝损伤发生发展相关的基因和关键信号通路。[方法]建立C57BL/6J小鼠APAP急性肝损伤模型。构建正常小鼠肝脏组织样本和APAP急性肝损伤小鼠损伤后第2天肝脏组织样本的2个cDNA文库,进行转录组测序。对获得的转录组测序数据进行组装及功能注释。使用DESeq R包(1.10.0)分析具有生物学重复的差异基因的表达,利用KOBAS软件确定KEGG信号通路中差异表达基因的静态富集。[结果]APAP急性肝损伤小鼠损伤后第2天与正常小鼠肝脏组织转录组相比,共有7270个DEGs,包括3707个显著(P<0.05)上调表达基因和3563个显著(P<0.05)下调表达基因。在所有显著上调表达基因中,表达量变化幅度较大的是Col1a1、Gsta1、S100a6基因;在所有显著下调表达基因中,差异表达量位于前3位的基因是Slc1a2、Glul、Acaa1b。共有2515个DEGs在316个不同的KEGG通路中富集,显著上调表达基因富集的信号通路主要是趋化因子信号通路、B细胞受体信号通路、NOD样受体信号通路、ECM受体相互作用信号通路等免疫和炎症相关信号通路,显著下调表达基因富集的信号通路主要是氧化磷酸化、脂肪酸降解、脂肪酸代谢、碳代谢等合成代谢信号通路。[结论]在APAP急性肝损伤过程中涉及多个信号通路的参与,趋化因子信号通路和ECM受体交互作用信号通路可能是急性损伤期中发挥主要作用的信号通路。

[Objective]To identify the key genes and signaling pathways associated with the occurrence and development of acetaminophen(APAP)-induced acute liver injury.[Method]An APAP-induced acute liver injury model was established in C57BL/6J mice.Two cDNA libraries from liver tissues of the healthy mice and the APAP-induced acute liver injury model mice on 2nd day of modeling were constructed and used to perform RNA-seq,respectively.The obtained transcriptome sequencing data were de novo assembled and functionally annotated.DESeq R package(1.10.0)was used for identification of differentially expressed genes(DEGs).KOBAS software was employed to conduct bioinformatics analysis on KEGG to determine the signaling pathways of DEGs enrichment associated with APAP-induced acute liver injury.[Result]Compared with the liver tissues of the healthy mice,a total of 7270 DEGs were found in those of the modeling mice on 2nd day of APAP-induced acute liver injury,of which 3707 DEGs were significantly(P<0.05)up-regulated and 3563 DEGs were significantly(P<0.05)down-regulated.Among the significantly up-regulated DEGs,Col1a1 was the most differentially expressed gene followed by Gsta1 and S100a6,and for the significantly down-regulated DEGs,the most differential expressions were observed in Slc1a2 followed by Glul and Acaa1b.Furthermore,a total of 2515 DEGs were enriched in 316 different KEGG pathways.The significantly up-regulated DEGs were enriched in the signaling pathways of chemokine,B lymphocytes receptor,NOD-like receptor,ECM receptor interaction and some immune and inflammatory signaling pathways,while the significantly down-regulated DEGs were enriched in those of oxidative phosphorylation,fatty acids degradation and metabolism,carbon metabolism and some synthetic and metabolic signaling pathways.[Conclusion]Multiple signaling pathways are involved in the process of APAP-induced acute liver injury,among which chemokine signaling pathway and ECM receptor interaction signaling pathway may exert leading roles in acute injury phase.