缺氧/复氧后鼠脑星形胶质细胞基因表达谱的初步分析

目的　该实验利用基因芯片技术研究缺氧 /复氧处理后鼠脑星形胶质细胞基因表达谱的变化 ,以深入认识星形胶质细胞在缺氧 /复氧处理后基因表达变化规律 ,为进一步全面认识神经胶质细胞在缺氧和缺血再灌注等情况下基因变化规律以及神经胶质细胞与神经元在损伤条件下相互依存关系提供实验依据和理论基础。方法　取人鼠脑星形胶质细胞作传代培养 ,传至第四代分组作缺氧 /复氧处理 ;用TRIzol试剂经过多个步骤提取制备好的星形胶质细胞样本总RNA ;利用基因芯片技术获取缺氧 /复氧处理后鼠脑星形胶质细胞的基因表达谱 ;利用GeneOntologyConsortium分析系统、OeneCards数据库和Pubmed检索系统对基因表达谱进行初步分析。结果　①基因表达谱 :在基因芯片测定了 4 0 96个点 ,共有差异表达基因 187个 ,其中 ,差异表达基因 187个 ,己知差异表达基因 4 6个 ,未知差异表达基因 14 1个 (EST片段 )。②已知差异表达基因 :在 4 6个已知差异表达基因中 ,表达下调的基因有 4 1个 ,表达上调的基因有 5个 ,未见报道的与缺氧 /复氧处理相关基因 2 3个 ,己报道的有 18个。③已知差异表达基因功能聚类 :共 10类 ,其中代谢 2 3个、信号传导 9个、细胞生长 3个、结构蛋白 2个、免疫应答 4个、运输 1个、细胞周期 1个、细胞增殖 1个

Primary analysis of gene expression profiles induced by hypoxia/reoxygenation in cultured cerebral astrocytes

Objective:To study the variety of gene expression profiles of rat cerebral astrocytes after hypoxia/reoxygenation (H/R) using microarray technology.Methods:Primary cultured cerebral astrocytes (40 days) were exposed to hypoxia (caused by mixed gases composing 95% N 2+5% CO 2)/reoxygenation. These cells were collected and total RNA samples were gained using TRIzol reagent. cDNA microarray chips comprising 4096 clones were employed to identify the effect of hypoxia/reoxygenation on gene expression profiles in cultured astrocytes. The obtained results were analyzed using Gene Ontology Consortium analysis system, GeneCards database and Pubmed Index system.Results:Gene expression profiles: There were differential expression gene 187 points in 4096 measured points, in which known differential expression gene 46 points and unknown differential expression gene 141 points (EST segment). Known differential expression gene: In the 46 points of known differential expression gene, there were down-regulate gene expression 41 points and up- regulate gene expression 5 points;twenty-eight genes related to hypoxia/reoxygenation had not been reported and 18 genes had been reported. Function of known differential expression gene: There were 10 kinds of it.Such as metabolism (23 points), signal transduction (9 points), cell growth (3 points), structure protein (2 points), immune response (4 points), transport (1 point), cell cycle (1 point), cell proliferation (1 point), apoptosis (1 point) and chaperone (1 point).Conclusions: The experiment achieved gene expression profile of cultured rat cerebral astrocytes following exposed to hypoxia/reoxygenation. It was the first discovery of 28 genes related to hypoxia/reoxygenation which had not been reported. The basic rule of differential expression of genes during hypoxia/ reoxygenation in cultured rat cerebral astrocytes was discovered primarily.