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王怡如,周唯,蒋笑影,潘一滨,许言午,曹永清.美沙拉秦干预下 IBD 小鼠结肠组织的转录组分析[J].中国实验动物学报,2022,30(1):7~16.
美沙拉秦干预下 IBD 小鼠结肠组织的转录组分析
RNA-seq based gene expression profiling of colon tissue from a mesalazine-treated mouse model of inflammatory bowel disease
投稿时间:2021-07-06  
DOI:10. 3969 / j.issn.1005-4847. 2022. 01. 001
中文关键词:  炎症性肠病  美沙拉秦  葡聚糖硫酸钠  转录组测序  差异表达基因
英文关键词:inflammatory bowel disease  mesalazine  dextran sodium sulfate  RNA sequencing  differentially expressed genes
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作者单位E-mail
王怡如 上海中医药大学附属龙华医院肛肠科,上海 200032 wangyiru1707@ 163.com 
周唯 上海中医药大学附属龙华医院肛肠科,上海 200032 zhouweitcm@ 163.com 
蒋笑影 上海中医药大学附属龙华医院肛肠科,上海 200032  
潘一滨 上海中医药大学附属龙华医院肛肠科,上海 200032  
许言午 上海中医药大学基础医学院生物化学教研室, 上海 201203  
曹永清 上海中医药大学附属龙华医院肛肠科,上海 200032 cyq88tcm@ 163.com 
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中文摘要:
       目的 应用转录组测序技术( RNA sequencing,RNA-seq) 检测炎症性肠病( inflammatory bowel disease,IBD)小鼠结肠组织的基因表达变化,增进对美沙拉秦(5-aminosalicylic acid,5-ASA)治疗 IBD 的分子机制的理解。 方法 用葡聚糖硫酸钠(dextran sulfate sodium,DSS)建立 IBD 小鼠模型。 将 15 只 C57BL/ 6 小鼠随机均分为三组:对照组、DSS 组和 DSS+5-ASA 组。 每组随机选择 3 只小鼠取结肠组织进行测序,经两组间比较,鉴定差异表达基因(differentially expressed genes,DEGs) 及重要生物学信号通路。 随后通过基因表达数据库( gene expression omnibus,GEO)中溃疡性结肠炎( ulcerative colitis,UC) 患者和经美沙拉秦治疗后患者结肠组织活检的微阵列 (microarray)数据,验证前面鉴定得到的子显著 DEGs。 结果 DSS+5-ASA 组与 DSS 组比较中共得到 2983 个差异表达基因,上调基因 604 个,下调基因 753 个;DSS 组与对照组比较,共得到 1626 个差异表达基因,其中上调基因 820 个,下调基因 806 个。 GO(gene ontology)和 KEGG 富集分析结果显示,这些 DEGs 与免疫反应等信号通路有关。部分表达差异显著的基因如 Rnase1 和 Cxcl10,在 UC 患者结肠组织中有相似的表达趋势。 结论 经美沙拉秦治疗的 IBD 模型小鼠结肠组织的基因表达谱,与 IBD 患者对比进行验证,得到数个 IBD 治疗过程中可能涉及的基因和生物学通路,增加了美沙拉秦对 IBD 治疗作用机制的进一步理解。
英文摘要:
       Objective To explore gene expression profiling of colon tissues from mesalazine-treated inflammatory bowel disease (IBD) mice using RNA-sequencing (RNA-seq), and provide a better understanding of the mechanism of mesalazine (5-ASA) in the treatment of IBD. Methods Dextran sodium sulfate (DSS) was used to establish the IBD mouse model. Fifteen mice (C57BL/ 6) were randomly assigned to three groups: control (n= 5), DSS (n= 5, using DSS to induce IBD), and DSS+5-ASA (n= 5, using 5-ASA to treat IBD). Three mice per group were randomly selected and colon tissues extracted for RNA-seq. Bioinformatic analysis was used for two comparisons (DSS vs. control, and DSS+5- ASA vs. DSS), and differentially expressed genes (DEGs) and biological signal pathways were identified. The top DEGs were further examined using microarray data of biopsies from ulcerative colitis patients and those treated with 5-ASA. Results Bioinformatic analysis identified 2983 DEGs, including 604 up-regulated and 753 down-regulated genes in the DSS+5-ASA group and DSS group comparison. In the DSS group and control group comparison, there were 1626 DEGs, 820 of which were up-regulated and 806 were down-regulated. Gene ontology and KEGG analysis of these DEGs suggested certain biological processes and pathways. The expression patterns of top DEG candidates, such as Rnase1 and Cxcl10, were confirmed in patients’ samples. Conclusions By exploring gene expression profiles in colon tissues from 5-ASA- treated IBD mice using RNA-seq and human mucosal biopsies, this study identified genes and pathways involved in the treatment process of IBD, providing new insights for understanding the mechanism of 5-ASA in IBD treatment.
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