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肠道缺血/再灌注损伤的分子机制：生物信息学分析与体内验证

Molecular Mechanisms Underlying Intestinal Ischemia/Reperfusion Injury: Bioinformatics Analysis and In Vivo Validation.

作者信息

Chen Fengshou, Wang Dan, Li Xiaoqian, Wang He

机构信息

Department of Anesthesiology, The First Hospital of China Medical University, Shenyang, Liaoning, China (mainland).

出版信息

Med Sci Monit. 2020 Dec 8;26:e927476. doi: 10.12659/MSM.927476.

DOI:10.12659/MSM.927476
PMID:33290384
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7733309/
Abstract

BACKGROUND Intestinal ischemia/reperfusion (I/R) injury is a serious clinical complication. This study aimed to explore the hub genes and pathways of intestinal I/R injury. MATERIAL AND METHODS GSE96733 from the GEO website was extracted to analyze the differentially expressed genes (DEGs) of intestinal I/R injured and sham-operated mice at 3 h and 6 h after surgery. The DAVID and STRING databases were used to construct functional enrichment analyses of DEGs and the protein-protein interaction (PPI) network. In Cytoscape software, cytoHubba was used to identify hub genes, and MCODE was used for module analysis. Testing by qRT-PCR detected the expression of hub genes in intestinal I/R injury. Western blot analysis detected the key proteins involved with the important pathways of intestinal I/R injury. RESULTS IL-6, IL-10, CXCL1, CXCL2, and IL-1ß were identified as critical upregulated genes, while IRF7, IFIT3, IFIT1, Herc6, and Oasl2 were identified as hub genes among the downregulated genes. The qRT-PCR testing showed the expression of critical upregulated genes was significantly increased in intestinal I/R injury (P<0.05), while the expression of hub downregulated genes was notably reduced (P<0.05). The proteins of CXCL1 and CXCR2 were upregulated following intestinal I/R injury (P<0.05) and the CXCL1/CXCR2 axis was involved with intestinal I/R injury. CONCLUSIONS The results of the present study identified IL-6, IL-10, CXCL1, CXCL2, IL-1ß, IRF7, IFIT3, IFIT1, Herc6, and Oasl2 as hub genes in intestinal I/R injury and identified the involvement of the CXCL1/CXCR2 axis in intestinal I/R injury.

摘要

背景 肠缺血/再灌注（I/R）损伤是一种严重的临床并发症。本研究旨在探索肠I/R损伤的核心基因和信号通路。

材料与方法 从基因表达综合数据库（GEO）网站提取GSE96733数据，以分析肠I/R损伤小鼠和假手术小鼠在术后3小时和6小时的差异表达基因（DEGs）。利用DAVID和STRING数据库对DEGs进行功能富集分析并构建蛋白质-蛋白质相互作用（PPI）网络。在Cytoscape软件中，使用cytoHubba识别核心基因，并使用MCODE进行模块分析。通过qRT-PCR检测肠I/R损伤中核心基因的表达。蛋白质印迹分析检测肠I/R损伤重要信号通路中涉及的关键蛋白。

结果 白细胞介素-6（IL-6）、白细胞介素-10（IL-10）、CXC趋化因子配体1（CXCL1）、CXC趋化因子配体2（CXCL2）和白细胞介素-1β（IL-1ß）被鉴定为关键上调基因，而干扰素调节因子7（IRF7）、干扰素诱导蛋白3（IFIT3）、干扰素诱导蛋白1（IFIT1）、含E3泛素蛋白连接酶结构域蛋白6（Herc6）和2'-5'-寡腺苷酸合成酶样蛋白2（Oasl2）被鉴定为下调基因中的核心基因。qRT-PCR检测显示，关键上调基因在肠I/R损伤中的表达显著增加（P<0.05），而核心下调基因的表达明显降低（P<0.05）。肠I/R损伤后，CXCL1和CXC趋化因子受体2（CXCR2）蛋白上调（P<0.05），且CXCL1/CXCR2轴与肠I/R损伤有关。

结论 本研究结果确定IL-6、IL-10、CXCL1、CXCL2、IL-1ß、IRF7、IFIT3、IFIT1、Herc6和Oasl2为肠I/R损伤的核心基因，并确定CXCL1/CXCR2轴参与肠I/R损伤。

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