基因芯片筛选颅内动脉瘤炎性相关基因的研究

Identification of inflammation-related genes in peripheral blood mononuclear cells from intracranial aneurysm patients using oligo-microarrays

目的采用基因芯片技术探讨颅内动脉瘤患者血液单核细胞中炎性相关基因的表达情况。方法采集50例颅内动脉瘤患者外周血,10名正常成人作为对照组,分离血液单核细胞,提取总RNA,与含22215个人类基因的寡核苷酸基因芯片杂交、洗脱、染色和扫描,分析检测的数据。荧光定量RT-PCR验证芯片结果。结果检测的22215个基因中,颅内动脉瘤共差异表达的基因有325个,差异水平均达2倍以上的基因有35个,生物信息学分析发现有16个基因与炎症反应和免疫应答有关,其中上调10个基因,下调6个。结论血液单核细胞基因表达谱是研究颅内动脉瘤发病机制的新策略,炎症反应和免疫应答可能是颅内动脉瘤形成和发展的重要机制。

Objective To identify the inflammation-related genes in peripheral blood mononuclear cells （PBMCs） from intracranial aneurysm patients using oligo-microarrays. Methods Blood samples from 50 intracranial aneurysms and 10 healthy human donors were obtained after informed consent. PBMC.,s were isolated from blood using FicoU method. The eDNA was retrotranscribed from the extracted RNA and the biotin-labeled cRNA derived from the transcription of eDNA was fragmented as probes. Then, the probes were hybridized with Affymetrix Human Genome U133A Array. Gene Array Scanner was used to screen the signals of hybridization and Microarray Suite software 5.0 was applied to detected the expression of genes. Results There are 325 genes differently expressed in PBMCs from intracranial aneurysm patients compared to healthy human. Function analysis were confirmed from 21 upregulated genes and 14 downregulated genes above 2-fold level. Among these genes, there were 16 genes involved in immune and inflammation. Conclusion The methodologies of genechip of PBMC.,s will provide a new powerful approach to help identify the molecular mechanisms of intracranial aneurysm. Immune and inflammation response may be the possible mechanism for the pathogenesis of intracranial aneurysms in Chinese.