大鼠心肌缺血预适应诱导表达上调基因的筛选和鉴定

Identification of Genes Up-regulated During Myocardial Ischemic Preconditiong in Rats

采用反复短时间结扎及松解左冠状动脉前降支构建大鼠心肌缺血预适应动物模型 ,运用抑制消减杂交技术建立大鼠心肌缺血预适应诱导表达上调基因的消减cDNA文库 ,通过反向RNA点杂交对部分文库基因进行差异表达初筛 ,选取表达差异最明显的 85个基因进行测序 ,获得了 3 1个核编码基因和 18个新基因 (EST) ,核编码基因中有相当一部分与细胞保护或信号转导有关 ,新基因已被GenBank收录 .从已测序基因中任选 5个进行RT PCR检测 ,并任选 2个进行RNA印迹检测 ,均证实在缺血预适应时表达增高 .

Brief periods of ischemia can protect the heart from a subsequent longer coronary artery occlusion, the phenomenon called ischemic preconditioning. The mechanism of ischemic preconditioning are not well understood A number of genes have been shown to play roles in cytoprotective effect of ishemic preconditioning, but there are clearly many missing elements to be found. Suppression subtractive hybridization was used to construct cDNA libraries enriched for genes up-regulated during ischemic preconditiong. After being confirmed by reverse Northern dot blot for differential expression, the selected genes were sequenced and searched in GenBank for homology analysis. Many nuclear-encoded genes that were up-regulated during ischemic preconditioning participate in cytoprotection. The 18 novel ESTs were banked into GenBank with accession numbers. The specificity of this response was confirmed by RT-PCR and Northern blot. Understanding the genes up-regulated during ischemic preconditioning may open new avenues for therapy in ischemic heart disease.