摘要
目的利用生物信息学方法对小鼠烧伤早期免疫细胞差异基因表达谱进行分析并检测,筛选相关差异基因,以期找到潜在的诊断及治疗靶标。方法从GEO数据库中下载GSE7404数据集,通过配对样本t检验、倍比法及GO功能富集分析筛选差异表达基因,应用STRING数据库及Cytoscape软件构建蛋白互作网络筛选相关基因靶标,运用实时荧光定量PCR及Western blotting进行试验验证。结果在烧伤后第1天,有1825个基因被选出,其中上调基因658个,下调基因1167个。运用STRING数据库及Cytoscape软件构建蛋白互作网络分析,发现Stat1、Cdk1、Cd19、Lck及Jun基因在烧伤后免疫系统功能变化过程中可能起到重要作用。实时荧光定量PCR及Western blotting检测显示Stat1、Cdk1和Jun基因表达情况与分析结果相一致。结论通过对小鼠烧伤后早期全血游离白细胞基因芯片分析我们发现Stat1、Cdk1和Jun基因在烧伤早期免疫细胞功能变化的过程中发挥重要的作用,可能是烧伤早期诊断及治疗的潜在的生物学靶标。
Objective To screen the genes related with leukocyte responses in mice early after burn injury by bioinformatic analysis of the gene expression profiling data. Methods The gene expression profiles were obtained from GEO (GSE7404, Mouse musculus, 25%TBSA, full-thickness) database. T test, fold changes and GO functional enrichment analysis were used to identify the differentially expressed genes (DEGs) related to leukocyte responses to burns; the interacting genes were transferred to STRING to construct the protein-protein interaction (PPI) network. Biological annotation of the sub-networks was executed using the software Cytoscape. Real-time PCR and Western blotting were used to verify the DEGs in mice. Results In mice at 1 day post-burn, a total of 658 genes were up-regulated and 1167 were down-regulated. PPI network and module analysis suggested that some of the genes (Stat1, Cdk1, Cd19, Lck and Jun) may play critical roles in the PPI network post-burn. Real-time PCR and Western blotting results in mice were consistent with those of bioinformatic analysis of Stat1, Cdk1 and Jun. Conclusion Stat1, Cdk1 and Jun might be critical players in the development of leukocyte response in mice early after burn injury. Our finding provides new insights into the pathogenesis of leukocyte response to burn injury and identifies several biomarkers as potential targets for burn treatment.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2017年第6期767-773,共7页
Journal of Southern Medical University
基金
广东省科技计划项目(2014A020212189)
南方医科大学南方医院院长基金(2014B013)
广州市科技计划项目(201607010242)
关键词
烧伤
免疫系统过程
差异表达基因
蛋白互作网络
post-burn injury
immune system process
differentially expressed genes
protein-protein interaction network