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[摘要]
目的：通过生物信息学方法分析地塞米松诱导的人小梁网基因表达谱变化，揭示激素性青光眼(GIG)发病过程中可能涉及的机制。
方法：从公共基因芯片数据库(GEO)获取基因表达数据集GSE37474，GEO2R筛选地塞米松组和正常小梁网之间的差异表达基因。使用DAVID数据库对差异基因进行GO功能注释和KEGG通路富集分析，使用STRING数据库和Cytoscape软件构建蛋白相互作用网络(PPI)，利用CytoHubba插件筛选关键枢纽基因，最后对关键枢纽基因的mRNA表达进行RT-PCR验证。
结果：与正常小梁网组织相比，地塞米松处理的小梁网组织有252个基因存在差异表达，其中141个为上调基因，111个为下调基因。GO功能注释显示差异基因主要定位于细胞外基质，参与炎症的正调控和细胞外基质重塑等生物学过程。KEGG通路富集分析显示差异基因主要参与血管平滑肌收缩、花生四烯酸代谢、醚脂质代谢和胆碱代谢。从PPI网络中筛选出7个关键枢纽基因，包括3个上调基因EDN1、FOS、LPL和4个下调基因CCL2、IGF1、PTGS2、CCL5，RT-PCR验证结果和基因表达谱芯片一致。
结论：地塞米松可引起小梁网基因表达谱发生显著变化，差异基因富集到的通路及筛选到的一些关键枢纽基因在细胞外基质重塑和房水流出调控中起着重要作用，这将有助于更全面地认识GIG的分子机制。
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[Abstract]
AIM:To profile gene expression changes induced by dexamethasone in human trabecular meshwork using bioinformatics analysis, and to elucidate the possible mechanisms involved in the pathogenesis of glucocorticoid-induced glaucoma(GIG).
METHODS:The gene expression dataset GSE37474 was obtained from the Gene Expression Omnibus(GEO). GEO2R was utilized to identify differentially expressed genes(DEGs)in trabecular meshwork between the dexamethasone group and the control group. Gene Ontology(GO)function annotation and the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were constructed using the DAVID database. STRING database and Cytoscape software were used to construct protein-protein interaction(PPI). The hub genes were screened by CytoHubba plug-in. Finally, the mRNA expression of key hub genes was verified by RT-PCR.
RESULTS:Compared to normal trabecular meshwork, dexamethasone-treated trabecular meshwork had a total of 252 DEGs, with 141 genes up-regulated and 111 genes down-regulated. GO function annotation showed that DEGs were mostly located in the extracellular matrix, where they engaged in the biological processes of positive regulation of inflammation and extracellular matrix remodeling. KEGG pathway enrichment showed that DEGs were largely involved in vascular smooth muscle contraction, arachidonic acid metabolism, ether lipid metabolism and choline metabolism. The PPI network yielded seven hub genes, three of which were up-regulated(EDN1, FOS, and LPL)and four of which were down-regulated(CCL2, IGF1, PTGS2, CCL5). In RT-PCR, the mRNA expression levels of the seven hub genes matched those in the gene expression profile.
CONCLUSION:Dexamethasone can cause dramatic changes in the gene expression profile in trabecular meshwork. The enriched pathways of DEGs and certain hub genes play an important role in the remodeling of the extracellular matrix and the regulation of aqueous humor outflow, providing a full knowledge of the molecular mechanism of GIG.
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