背景:氧化应激(OS)在缺氧缺血性脑损伤(HIBD)的进展中起着重要作用。本研究旨在探讨新生儿HIBD中OS相关基因及其分子机制。
方法:从基因表达综合(GEO)数据库获得微阵列数据集,以筛选对照样品和HIBD样品之间的差异表达基因(DEG)。从GeneCards中提取OS相关基因,并通过与DEG相交获得HIBD中的OS-DEG。随后,基因本体论(GO)和京都基因和基因组百科全书(KEGG),进行基因集富集分析(GSEA)以确定OS-DEG在HIBD中的潜在机制和功能。此外,使用蛋白质-蛋白质相互作用网络筛选hub基因,并在GSE144456数据集中鉴定.然后进行CIBERSORT以评估每个样品中免疫细胞的表达并进行最佳OS-DEG和免疫细胞的相关性分析。最后,进行定量逆转录聚合酶链反应(RT-qPCR)和免疫组织化学以验证最佳OS-DEGs的表达水平。
结果:总计,确定了93个OS-DEG。GO,KEGG,和GSEA富集分析表明,这些基因主要富集在OS和炎症中。四个OS相关的生物标志物基因(Jun,Fos,Tlr2和Atf3)被鉴定和验证。CIBERSORT分析揭示了HIBD组中六种类型的免疫细胞的失调。此外,筛选了47种可能靶向4种OS相关生物标志物基因的药物。最终,RT-qPCR和免疫组化结果进一步验证了Fos的表达水平,Tlr2和Atf3。
结论:Fos,Tlr2和Atf3是HIBD进展的潜在OS相关生物标志物。OS的机制与新生儿HIBD的机制有关。
BACKGROUND: Oxidative stress (OS) plays a major role in the progress of hypoxic-ischemic brain damage (HIBD). This study aimed to investigate OS-related genes and their underlying molecular mechanisms in neonatal HIBD.
METHODS: Microarray data sets were acquired from the Gene Expression Omnibus (GEO) database to screen the differentially expressed genes (DEGs) between control samples and HIBD samples. OS-related genes were drawn from GeneCards and OS-DEGs in HIBD were obtained by intersecting with the DEGs. Subsequently, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA) were conducted to determine the underlying mechanisms and functions of OS-DEGs in HIBD. Moreover, the hub genes were screened using the protein-protein interaction network and identified in the GSE144456 data set. CIBERSORT was then performed to evaluate the expression of immunocytes in each sample and perform a correlation analysis of the optimal OS-DEGs and immunocytes. Finally, quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunohistochemistry were performed to validate the expression levels of the optimal OS-DEGs.
RESULTS: In total, 93 OS-DEGs were identified. GO, KEGG, and GSEA enrichment analyses indicated that these genes were predominantly enriched in OS and inflammation. Four OS-related biomarker genes (Jun, Fos, Tlr2, and Atf3) were identified and verified. CIBERSORT analysis revealed the dysregulation of six types of immune cells in the HIBD group. Moreover, 47 drugs that might target four OS-related biomarker genes were screened. Eventually, RT-qPCR and immunohistochemistry results for rat samples further validated the expression levels of Fos, Tlr2, and Atf3.
CONCLUSIONS: Fos, Tlr2 and Atf3 are potential OS-related biomarkers of HIBD progression. The mechanisms of OS are associated with those of neonatal HIBD.