实验性蛛网膜下腔出血后脑动脉的基因表达谱及功能分析

Gene expression profiling and functional analysis of cerebral artery after experimental subarachnoid hemorrhage

目的探讨兔蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)基底动脉和正常基底动脉基因表达的差异。方法兔SAH模型脑基底动脉及正常兔脑基底动脉的c DNA基因芯片下载于GEO数据库。使用Bioconductor软件对芯片进行分析筛选,并用Cytoscape软件对差异表达基因进行功能富集和信号通路分析。随后选取成年雄性日本大耳兔6只,随机分为正常对照组(n=3)和SAH模型组(n=3)。采用枕大池二次注血法复制兔SAH模型,取第0天未行手术处理的对照兔基底动脉标本RNA和第5天SAH后基底动脉标本RNA行qRT-PCR,验证部分差异基因。结果在兔正常基底动脉和兔SAH模型基底动脉中共获得4 356个差异表达的基因,其中920个差异表达基因(P<0.05),如GRIK1、MYH13、ZNF45、SAA3、RLN1、MSR1等。功能富集分析结果显示差异表达基因涉及钙离子跨膜转运蛋白活性调节、离子跨膜转运负调控、钾离子转运调控、JAK-STAT信号通路级联的正调节等相关生物学过程。通路分析显示这些基因与钙信号通路、cGMP-PKG信号通路、HIF-1信号通路、PI3K-Akt信号通路等有关。qRT-PCR验证表明SAH模型组MSR1下调,与芯片结果一致。结论兔造模后CVS基底动脉中存在基因的差异化表达,并且MSR1基因可以作为研究CVS病理机制的潜在靶点。

Objective To explore the difference of gene expression profiling between normal basilar arteries and basilar arteries of cerebral vasospasm(CVS)after subarachnoid hemorrhage(SAH)in rabbits.Methods cDNA chip of normal basilar arteries and basilar arteries of CVS after SAH in rabbits were downloaded from GEO database.The chip was analyzed and screened by Bioconductor software,and function enrichment and pathway analysis of the differentially expressed genes were analyzed by Cytoscape software.Then6adult male Japanese rabbits were used,and randomly divided into normal control group(n=3)and SAH model group(n=3).Rabbit SAH models were established by cisterna secondaryblood-injection method.RNA data of normal basilar artery specimens on the0day and basilar artery specimens after SAH on the5-day were used to validate the parts of differentially expressed genes by qRT-PCR.Results A total of4356differentially expressed genes were found in normal basilar arteries and basilar arteries of CVS after SAH in rabbits.Among them,920genes were considered to be significant with P-value<0.05,such as GRIK1,MYH13,ZNF45,SAA3,RLN1,MSR1and others.Function enrichment analysis indicated that the differentially expressed genes were involved in regulationof Ca2+transmembrane transporter activity,negative regulation of ion transmembrane transport,regulation of potassium iontransport,positive regulation of JAK-STAT signaling cascades and other biological processes.Pathway analysis showed that calcium signaling pathway,cGMP-PKG signaling pathway,HIF-1signaling pathway,PI3K-Akt signaling pathway andother signaling pathways maybe related with the differentially expressed genes.qRT-PCR verification showed that the expression of MSR1in SAH model group was consistent with that of the chip result.Conclusion The gene expressions of basilar arteries of CVS after SAH in rabbits are significantly different,and MSR1gene can be used as a potential target for studying the pathological mechanism of CVS.