摘要
目的检测远隔缺血预处理对大鼠缺血半暗带脑组织基因表达的影响,探讨远隔缺血预处理诱导的内源性脑保护机制。方法雄性SD大鼠24只随机分为常规缺血组(n=12)和远隔预处理组(n=12)。常规缺血组应用远端大脑中动脉阻塞术(dMCAO)建立局灶性脑缺血模型;远隔预处理组给予左侧股动脉15 min夹闭/15 min再通3个循环缺血预处理后,再予以dMCAO。两组大鼠分别在缺血1、3、6和24 h后取脑,采用Agilent大鼠全基因组芯片检测两组脑缺血半暗带区基因表达的变化,获取差异表达基因,并对差异基因进行GO分析和Pathway功能注释(KEGG数据库)。结果远隔缺血预处理后:①缺血半暗带区脑组织基因出现差异表达(P<0.05且差异倍数>2),随着缺血时间延长差异基因个数逐渐增加,在缺血6 h达高峰;②GO分析结果提示差异基因涉及刺激反应、生长、生物调节、细胞杀伤等多个生物过程;③对差异基因进行KEGG通路分析,结果提示Jak-STAT信号通路、p53信号通路、丝裂原蛋白激酶(MAPK)信号通路、谷氨酸突触信号通路、血管内皮生长因子(VEGF)信号通路等多条分子通路出现差异调节,差异调节通路的数目也在缺血6 h达高峰。结论远隔缺血预处理诱导半暗带基因差异表达,差异基因通过多条分子通路影响多个生物过程。基因表达谱芯片为远隔缺血预处理脑保护机制提供了全面的数据和信息,对各个靶点、各条通路的进一步深入研究具有全面、广泛的指导意义。
Aim To explore the effect of limb remote ischemic preconditioning（RIPC） on gene expression of ischemic penumbra after focal cerebral ischemia in rats.Methods A total of 24 male Sprague-Dawley rats were randomly assigned to a control ischemia group（n=12） and a RIPC group（n=12）.Rats in the control ischemia group were subjected to focal cerebral ischemia induced by the distal middle cerebral artery occlusion（dMCAO） method.Rats in the RIPC group were treated with three cycles of 15 min occlusion/reperfusion of the left femoral artery before dMCAO.The rats＇ brains were harvested 1 h,3 h,6 h and 24 h after cerebral ischemia,and gene expression microarrays were used to identify the differently expressed genes.Then GO（gene ontology） functional annotation and KEGG pathway analysis were used to explore the neuroprotection mechanisms of limb remote ischemic preconditioning.Results RIPC regulated the expression levels of many genes in ischemic penumbra.The number of differently expressed genes increased progressively,and reached the maximum at 6 h after ischemia.GO analysis indicated that the differently expressed genes were related to biological regulation,response to stimulus,growth,cell killing,and so on.KEGG pathway analysis revealed that activity mainly was related to Jak-STAT signaling pathway,p53 signaling pathway,MAPK signaling pathway,glutamatergic synapse,VEGF signaling pathway,and so on.The number of differently regulated pathways also reached the maximum at 6 h after ischemia.Conclusion RIPC regulated the expression levels of many genes in ischemic penumbra.The differently expressed genes were involved many biological process and signaling pathways.These results provided a great deal of useful information and would help explore the neuroprotection mechanisms of limb remote ischemic preconditioning.
出处
《中国临床神经科学》
2015年第1期1-8,共8页
Chinese Journal of Clinical Neurosciences
基金
国家自然科学基金(编号:81071061)
上海市医学重点专科课题(编号:ZK2012B02-1)
上海市自然科学基金(编号:10411968900)
关键词
远隔缺血预处理
脑保护
基因表达谱
大鼠
remote ischemic preconditioning
neuroprotection
gene expression profile
rat
