| 摘要: |
| 目的:研究脂多糖(LPS)诱导的脓毒症小鼠脾脏中mRNA表达变化及特征，寻找影响脓毒症病理生理进程的信号通路。方法:将50只小鼠分成LPS组和Sham组，每组25只，比较两组小鼠存活率及血清中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6和IL-1β水平。提取脾脏中mRNA，利用转录组测序技术测定基因表达量，随后通过两种富集方法分析:寻找差异表达基因(P< 0.05, |log2FoldChange| ≥1)进行GO和KEGG分析以及在GSEA v4.0.1软件中富集P < 0.05，Q < 0.25的基因。RT-qPCR验证10个显著差异表达基因。结果: LPS成功构建脓毒症模型，表现出严重炎症反应和高死亡率。LPS组对比Sham组显著改变的差异表达基因有1 616个，其中下调基因919个，上调基因697个。富集分析涉及多条炎症反应、调控遗传信息、参与信号转导和细胞过程的通路。RT-qPCR验证结果显示10个基因表达趋势与测序结果一致。结论:胞质DNA传感途径可能在脓毒症病理过程中起着重要作用，Zbp1、Ddx58、Cxcl10、Polr1d可能是其发挥功能的重要靶点。 |
| 关键词: 脓毒症 转录组测序 mRNA 基因表达 |
| DOI：10.3969/j.issn.1007-6948.2022.04.018 |
| 投稿时间：2021-11-28 |
| 基金项目:国家自然科学基金(81774108)；上海市卫健委科研课题计划(201940062) |
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| Analysis of Differentially Expressed Genes in the Spleen of Sepsis Mice Based on RNA-Seq |
| LU Liyue,YONG Yue,GAO Hao |
| Department of Anesthesiology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China |
| Abstract: |
| Objective To detect the changes and characteristics of mRNA expression in the spleen of sepsis mice induced by LPS and look for the signaling pathways that affect the pathophysiological process of sepsis. Methods The mice were divided into LPS and Sham group, and the mortality and serum levels of TNF-α, IL-6 and IL-1β were observed to evaluate the model. Extract RNA from the spleens of mice, RNA-Seq to determine gene expression and then analyze by two methods: Differentially expressed genes (P<0.05, |log2 Fold Change|≥1) were screened. GO and KEGG were performed and Enrichment of genes with P<0.05 and Q<0.25 in GSEA v4.0.1 software. RT-qPCR verified the 10 differentially expressed genes in the RNA-Seq results. Results LPS successfully constructed a sepsis model, which showed severe inflammation and high mortality (P<0.01). Compared with the Sham group, 1616 differentially expressed genes were changed in the LPS group, of which 919 were down-regulated genes and 697 were up-regulated genes. GO and KEGG analysis involve multiple pathways of inflammation, regulation of genetic information, signal transduction, cellular processes.The RT-qPCR verification results showed that the expression trends of 10 genes were consistent with the sequencing results. Conclusion The cytoplasmic DNA sensing pathway may play an important role in the pathological process of sepsis. Zbp1, Ddx58, Cxcl10 and Polr1d may be important targets for their functions. |
| Key words: sepsis transcriptome sequencing mRNA gene expression |
