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| 基于mRNA-seq数据筛选Mn2＋调控小鼠巨噬细胞的差异表达基因 |
| 吴双双,陈晶,王云成,马一瑄,魏书宇,刘鸿雁,董亚尊,王北艳,崔玉东,马金柱* |
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(黑龙江八一农垦大学生命科学技术学院, 大庆 163319
*通信作者) |
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| 摘要: |
| 目的 对Mn2＋调控的小鼠巨噬细胞mRNA进行测序和生物信息学分析，筛选出差异表达的基因并分析其生物学功能，探究Mn2＋对小鼠巨噬细胞的调控作用。方法 采用MnCl2和NaCl分别处理小鼠腹腔巨噬细胞24 h，利用mRNA-seq方法筛选出Mn2＋调控巨噬细胞的差异表达基因，对差异表达基因进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析，并采用qPCR对关键的差异表达基因进行验证。结果 通过整合数据库信息和分析获得1 637个差异表达基因，包括表达上调的基因912个、表达下调的基因725个，GO富集分析表明这些差异表达基因具有免疫吞噬、细胞增殖和分化、转录因子调控等生物活性，KEGG通路主要富集在PI3K-Akt、NF-κB等信号通路。其中与免疫相关且差异最显著的10个基因为免疫球蛋白G Fc段受体Ⅰ（Fcgr1）、含不育α基序结构域蛋白11（Samd11）、活性调节的细胞骨架相关蛋白（Arc）、G蛋白偶联受体35（Gpr35）、富含脯氨酸的小蛋白2H（Sprr2h）、Wnt家族成员4（Wnt4）、整合膜蛋白2A（Itm2a）、无调性bHLH转录因子8（Atoh8）、泛素结合酶E2C（Ube2c）及富含脯氨酸的小蛋白2B（Sprr2b），qPCR验证结果与mRNA测序分析趋势一致。结论 通过mRNA-seq数据筛选Mn2＋调控小鼠巨噬细胞的差异表达基因及GO和KEGG富集分析结果表明Mn2＋对巨噬细胞具有免疫调节作用。 |
| 关键词: mRNA测序 巨噬细胞 锰 免疫调节 差异表达基因 |
| DOI：10.16781/j.CN31-2187/R.20210989 |
| 投稿时间：2021-09-28修订日期：2021-12-07 |
| 基金项目:黑龙江省自然科学基金联合引导项目(LH2019C047),大庆市指导性科技项目(zd-2020-62),留学归国科研启动基金计划(ZRCLG201905),黑龙江八一农垦大学研究生创新科研项目(YJSCX2021-Y104),大学生创新创业训练计划(202110223005,202110223007). |
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| Screening of differentially expressed genes in mouse macrophages regulated by Mn2＋ based on mRNA-seq data |
| WU Shuang-shuang,CHEN Jing,WANG Yun-cheng,MA Yi-xuan,WEI Shu-yu,LIU Hong-yan,DONG Ya-zun,WANG Bei-yan,CUI Yu-dong,MA Jin-zhu* |
(College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, Heilongjiang, China
*Corresponding author) |
| Abstract: |
| Objective To conduct sequencing and bioinformatics analysis of mRNA from mouse macrophages regulated by Mn2＋, screen out differentially expressed genes, and analyze their biological functions, so as to explore the regulatory effect of Mn2＋ on mouse macrophages. Methods MnCl2 and NaCl were used to treat mouse peritoneal macrophages for 24 h, respectively. The mRNA-seq method was used to screen for the differentially expressed genes of macrophages regulated by Mn2＋, then the differentially expressed genes were analyzed with the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and the key differentially expressed genes were verified by quantitative polymerase chain reaction (qPCR). Results A total of 1 637 differentially expressed genes were obtained by integrating database information and analyses, including 912 upregulated genes and 725 down-regulated genes. GO enrichment analysis showed that these differentially expressed genes had biological activities, such as phagocytosis, cell proliferation and differentiation, and regulation of transcription factors. The KEGG pathway was mainly enriched in phosphatidylinositol 3-kinase-protein kinase B and nuclear factor κB signaling pathways. The top 10 genes associated with immunity and with the most significant differences were Fc receptor, immunoglobulin G, high affinity Ⅰ (Fcgr1), sterile α motif domain containing 11 (Samd11), activity regulated cytoskeleton-associated protein (Arc), G protein-coupled receptor 35 (Gpr35), small proline-rich protein 2H (Sprr2h), Wnt family member 4 (Wnt4), integral membrane protein 2A (Itm2a), atonal bHLH transcription factor 8 (Atoh8), ubiquitin conjugating enzyme E2C (Ube2c) and small proline rich protein 2B (Sprr2b). The qPCR verification results of the 10 genes were consistent with the trend of mRNA sequencing analysis. Conclusion The screening of the differentially expressed genes from mouse macrophages regulated by Mn2＋ based on mRNA-seq data and the analyses of GO and KEGG enrichment show that Mn2＋ has an immunoregulation effect on mouse macrophages. |
| Key words: mRNA sequencing macrophages manganese immunoregulation differentially expressed genes |
