脓毒症相关肠黏膜屏障损伤关键基因和信号通路的转录组学分析
Transcriptomic Analysis of Key Genes and Signaling Pathways in Sepsis-Associated Intestinal Mucosal Barrier Damage
DOI: 10.12677/acm.2024.1451707, PDF,   
作者: 高 昭, 高 鹏*：青岛大学附属医院急诊外科，山东 青岛；黄 海：常州市武进人民医院急诊科，江苏 常州
关键词: 脓毒症；肠道屏障；差异表达基因；转录组测序；Sepsis； Intestinal Barrier； Differentially Expressed Genes； Transcriptome Sequencing
摘要: 目的：分析脓毒症相关肠黏膜屏障损伤小鼠差异表达基因(DEG)，从转录组水平探讨脓毒症相关肠黏膜屏障损伤的早期诊断和保护机制。方法：收集行盲肠结扎穿孔术(CLP)和假手术的健康雄性C57BL/6J小鼠小肠组织，采用illumina Hiseq测序平台的两端测序模式进行高通量测序。对差异表达基因进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)分析。利用STRING数据库构建蛋白–蛋白相互作用(PPI)网络，利用Cytoscape筛选关键基因。采用实时荧光定量聚合酶链反应(RT-qPCR)对Hub基因进行验证。结果：共筛选出239个DEG，其中上调基因49个，下调基因130个。KEGG富集分析发现DEG主要涉及细胞因子–细胞因子受体相互作用、Th1和Th2细胞分化、病毒蛋白与细胞因子和细胞因子受体相互作用、IL-17信号通路等。利用cytoHubba插件筛选出前10个hub基因。经实验验证，TBX21、CSF3、IL-6、CXCR3、CXCL9的表达与测序结果一致。结论：TBX21、CSF3、IL-6、CXCR3和CXCL9可能是脓毒症相关肠黏膜屏障早期诊断和治疗的潜在生物学标志物。
Abstract: Objectives: To analyze the differentially expressed genes (DEG) in mice with sepsis-related intestinal mucosal barrier damage and to explore the early diagnosis and protection mechanism of sepsis-related intestinal mucosal barrier damage at the transcriptome level. Methods: The healthy male C57BL/6J mice small intestinal tissues of Cecal ligation and puncture (CLP) and sham-operation were collected, and the two-end sequencing mode of the illumina Hiseq sequencing platform was commissioned for high-throughput sequencing. Followed by Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of DEGs. A protein-protein interaction (PPI) network was constructed using the STRING database, and hub genes were selected using Cytoscape. Hub genes were verified using quantitative real-time polymerase chain reaction (RT-qPCR). Results: A total of 239 DEG were selected, including 49 up-regulated genes and 130 downregulated genes. KEGG enrichment analysis revealed that DEG mainly involved cytokine-cytokine receptor interaction, Th1 and Th2 cell differentiation, viral protein interaction with cytokine and cytokine receptor, and IL-17 signaling pathway. The cyto Hubba plugin was used to select the top 10 hub genes. After experimental verification, the expression of TBX21, CSF3, IL-6, CXCR3, and CXCL9 was in agreement with the sequencing results. Conclusion: TBX21, CSF3, IL-6, CXCR3, and CXCL9 may be potential biological markers for the early diagnosis and treatment the sepsis-associated intestinal mucosal barrier.
文章引用：高昭, 黄海, 高鹏. 脓毒症相关肠黏膜屏障损伤关键基因和信号通路的转录组学分析[J]. 临床医学进展, 2024, 14(5): 2457-2470. https://doi.org/10.12677/acm.2024.1451707

参考文献

[1] Evans, L., Rhodes, A., Alhazzani, W., et al. (2021) Surviving Sepsis Campaign: International Guidelines for Management of Sepsis and Septic Shock 2021. Intensive Care Medicine, 47, 1181-1247. [Google Scholar] [CrossRef] [PubMed]
[2] Van Der Poll, T., Shankar-Hari, M. and Wiersinga, W.J. (2021) The Immunology of Sepsis. Immunity, 54, 2450-2464. [Google Scholar] [CrossRef] [PubMed]
[3] Weng, L., Zeng, X.Y., Yin, P., et al. (2018) Sepsis-Related Mortality in China: A Descriptive Analysis. Intensive Care Medicine, 44, 1071-1080. [Google Scholar] [CrossRef] [PubMed]
[4] Carrico, C.J., Meakins, J.L., Marshall, J.C., et al. (1986) Multiple-Organ-Failure Syndrome. Archives of Surgery, 121, 196-208. [Google Scholar] [CrossRef] [PubMed]
[5] Rudd, K.E., Johnson, S.C., Agesa, K.M., et al. (2020) Global, Regional, and National Sepsis Incidence and Mortality, 1990-2017: Analysis for the Global Burden of Disease Study. The Lancet, 395, 200-211. [Google Scholar] [CrossRef] 
[6] Mowat, A.M. and Agace, W.W. (2014) Regional Specialization within the Intestinal Immune System. Nature Reviews Immunology, 14, 667-685. [Google Scholar] [CrossRef] [PubMed]
[7] Stadnyk, A.W. (2002) Intestinal Epithelial Cells as a Source of Inflammatory Cytokines and Chemokines. Canadian Journal of Gastroenterology, 16, 241-246. [Google Scholar] [CrossRef] [PubMed]
[8] Verdu, E.F., Galipeau, H.J. and Jabri, B. (2015) Novel Players in Coeliac Disease Pathogenesis: Role of the Gut Microbiota. Nature Reviews Gastroenterology & Hepatology, 12, 497-506. [Google Scholar] [CrossRef] [PubMed]
[9] Paucar Iza, Y.A. and Brown, C.C. (2024) Early Life Imprinting of Intestinal Immune Tolerance and Tissue Homeostasis. Immunological Reviews, 323, 303-315. [Google Scholar] [CrossRef] [PubMed]
[10] Uhle, F., Lichtenstern, C., Brenner, T., et al. (2015) Pathophysiology of Sepsis. Anästhesiologie, Intensivmedizin, Notfallmedizin, Schmerztherapie, 50, 114-122. [Google Scholar] [CrossRef] [PubMed]
[11] Zhou, Q. and Verne, G.N. (2018) Intestinal Hyperpermeability: A Gateway to Multi-Organ Failure? Journal of Clinical Investigation, 128, 4764-4766. [Google Scholar] [CrossRef] 
[12] Rittirsch, D., Huber-Lang, M.S., Flierl, M.A., et al. (2009) Immunodesign of Experimental Sepsis by Cecal Ligation and Puncture. Nature Protocols, 4, 31-36. [Google Scholar] [CrossRef] [PubMed]
[13] Chiu, C.J., Mcardle, A.H., Brown, R., et al. (1970) Intestinal Mucosal Lesion in Low-Flow States. I. A Morphological, Hemodynamic, and Metabolic Reappraisal. Archives of Surgery, 101, 478-483. [Google Scholar] [CrossRef] [PubMed]
[14] Rowlands, B.J., Soong, C.V. and Gardiner, K.R. (1999) The Gastrointestinal Tract as a Barrier in Sepsis. British Medical Bulletin, 55, 196-211. [Google Scholar] [CrossRef] [PubMed]
[15] Mazarati, A., Medel-Matus, J.S., Shin, D., et al. (2021) Disruption of Intestinal Barrier and Endotoxemia after Traumatic Brain Injury: Implications for Post-Traumatic Epilepsy. Epilepsia, 62, 1472-1481. [Google Scholar] [CrossRef] [PubMed]
[16] Haussner, F., Chakraborty, S., Halbgebauer, R., et al. (2019) Challenge to the Intestinal Mucosa during Sepsis. Frontiers in Immunology, 10, Article No. 891. [Google Scholar] [CrossRef] [PubMed]
[17] Lu, L., Liu, L.P., Gui, R., et al. (2022) Discovering Common Pathogenetic Processes between COVID-19 and Sepsis by Bioinformatics and System Biology Approach. Frontiers in Immunology, 13, Article ID: 975848. [Google Scholar] [CrossRef] [PubMed]
[18] Russell, J.A. (2006) Management of Sepsis. The New England Journal of Medicine, 355, 1699-1713. [Google Scholar] [CrossRef] 
[19] Delano, M.J., Scumpia, P.O., Weinstein, J.S., et al. (2007) MyD88-Dependent Expansion of an Immature GR-1( )CD11b( ) Population Induces T Cell Suppression and Th2 Polarization in Sepsis. Journal of Experimental Medicine, 204, 1463-1474. [Google Scholar] [CrossRef] [PubMed]
[20] Geng, F., Liu, W. and Yu, L. (2022) MicroRNA-451a and Th1/Th2 Ratio Inform Inflammation, Septic Organ Injury, and Mortality Risk in Sepsis Patients. Frontiers in Microbiology, 13, Article ID: 947139. [Google Scholar] [CrossRef] [PubMed]
[21] Delano, M.J., Thayer, T., Gabrilovich, S., et al. (2011) Sepsis Induces Early Alterations in Innate Immunity That Impact Mortality to Secondary Infection. The Journal of Immunology, 186, 195-202. [Google Scholar] [CrossRef] [PubMed]
[22] Condotta, S.A., Khan, S.H., Rai, D., et al. (2015) Polymicrobial Sepsis Increases Susceptibility to Chronic Viral Infection and Exacerbates CD8 T Cell Exhaustion. The Journal of Immunology, 195, 116-125. [Google Scholar] [CrossRef] [PubMed]
[23] Ahmed Ali, M., Mikhael, E.S., Abdelkader, A., et al. (2018) Interleukin-17 as a Predictor of Sepsis in Polytrauma Patients: A Prospective Cohort Study. European Journal of Trauma and Emergency Surgery, 44, 621-626. [Google Scholar] [CrossRef] [PubMed]
[24] Cua, D.J. and Tato, C.M. (2010) Innate IL-17-Producing Cells: The Sentinels of the Immune System. Nature Reviews Immunology, 10, 479-489. [Google Scholar] [CrossRef] [PubMed]
[25] Nakada, T.A., Russell, J.A., Boyd, J.H., et al. (2011) IL17A Genetic Variation Is Associated with Altered Susceptibility to Gram-Positive Infection and Mortality of Severe Sepsis. Critical Care, 15, R254. [Google Scholar] [CrossRef] [PubMed]
[26] Song, X., Zhu, S., Shi, P., et al. (2011) IL-17RE Is the Functional Receptor for IL-17C and Mediates Mucosal Immunity to Infection with Intestinal Pathogens. Nature Immunology, 12, 1151-1158. [Google Scholar] [CrossRef] [PubMed]
[27] Ge, Y., Huang, M. and Yao, Y.M. (2020) Biology of Interleukin-17 and Its Pathophysiological Significance in Sepsis. Frontiers in Immunology, 11, Article No. 1558. [Google Scholar] [CrossRef] [PubMed]
[28] Ogiku, M., Kono, H., Hara, M., et al. (2012) Interleukin-17A Plays a Pivotal Role in Polymicrobial Sepsis According to Studies Using IL-17A Knockout Mice. Journal of Surgical Research, 174, 142-149. [Google Scholar] [CrossRef] [PubMed]
[29] Hotchkiss, R.S., Monneret, G. and Payen, D. (2013) Sepsis-Induced Immunosuppression: From Cellular Dysfunctions to Immunotherapy. Nature Reviews Immunology, 13, 862-874. [Google Scholar] [CrossRef] [PubMed]
[30] Ma, L., Li, Q., Cai, S., et al. (2021) The Role of NK Cells in Fighting the Virus Infection and Sepsis. International Journal of Medical Sciences, 18, 3236-3248. [Google Scholar] [CrossRef] [PubMed]
[31] Frydrych, L.M., Bian, G., Fattahi, F., et al. (2019) GM-CSF Administration Improves Defects in Innate Immunity and Sepsis Survival in Obese Diabetic Mice. The Journal of Immunology, 202, 931-942. [Google Scholar] [CrossRef] [PubMed]
[32] Almansa, R., Heredia-Rodríguez, M., Gomez-Sanchez, E., et al. (2015) Transcriptomic Correlates of Organ Failure Extent in Sepsis. Journal of Infection, 70, 445-456. [Google Scholar] [CrossRef] [PubMed]
[33] Liang, J., Wu, W., Wang, X., et al. (2022) Analysis of Sepsis Markers and Pathogenesis Based on Gene Differential Expression and Protein Interaction Network. Journal of Healthcare Engineering, 2022, Article ID: 6878495. [Google Scholar] [CrossRef] [PubMed]
[34] De Matteis, A., Colucci, M., Rossi, M.N., et al. (2022) Expansion of CD4dimCD8 T Cells Characterizes Macrophage Activation Syndrome and Other Secondary HLH. Blood, 140, 262-273. [Google Scholar] [CrossRef] [PubMed]
[35] Danahy, D.B., Anthony, S.M., Jensen, I.J., et al. (2017) Polymicrobial Sepsis Impairs Bystander Recruitment of Effector Cells to Infected Skin despite Optimal Sensing and Alarming Function of Skin Resident Memory CD8 T Cells. PLOS Pathogens, 13, e1006569. [Google Scholar] [CrossRef] [PubMed]

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