摘要
目的 检测难治性癫痫鼠脑细胞信号转导相关基因的表达 ,探讨难治性癫痫的发病机理。方法 建立难治性癫痫和非难治性癫痫大鼠模型 ,抽提两者脑组织的RNA ,分别逆转录合成荧光分子掺入的cDNA做探针 ;将含有 4 0 96条人类基因PCR产物制成基因芯片 ,芯片杂交和严格洗片后 ,用ScanArray30 0 0荧光扫描仪扫描芯片荧光信号图像 ,利用计算机分析难治性和非难治性癫痫鼠脑差异表达的基因。结果 在 4 0 96种基因中 ,难治性和非难治性癫痫鼠脑组织间存在明显差异表达的基因 36 4条 ,其中与细胞信号转导相关的基因有 2 9条 ,上调的有 1 0条 ,下调的有 1 9条。结论 微矩阵基因芯片在筛选难治性癫痫相关基因的改变时 ,具有快速、高通量、高灵敏度等特点 。
Objective To explore the pathogenic mechanisms of intractable epilepsy by searching for the differential cell signal transduction associated genes expression in intractable and non intractable epilepsy rat brain using cDNA microarray Methods Intractable epilepsy and non intractable epilepsy rat model were build The total RNAs were isolated from the brain tissues Both mRNAs from the brains of the intractable and non intractable epilepsy rats were reversely transcribed to the cDNA with the incorporation of fluorescent dUTP to prepare the hybridization probes The PCR products of 4 096 human genes were spotted on a chemical material coated glass plates in array The mixed probes were then hybridized to the cDNA microarray After high stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed differences between 2 tissues Results Among the 4 096 target genes, 29 genes associated with cell signal transduction differentially expressed were identified, 10 were up regulated(34 48%) and 19 down regulated(65 52%) Conclusion cDNA microarray technology is an effective technique in screening the differentially expressed genes between intractable and non intractable epilepsy rat brain Disturbances of cell signal transduction play a role in the pathogenic mechanism of intractable epilepsy
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2004年第3期171-174,共4页
Chinese Journal of Laboratory Medicine
