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脓毒症大鼠心脏组织基因表达变化的研究

摘要

Objective To investigate the changes in gene expression spectrum of heart tissue in septic rats by DNA microarrays. Methods Thirty male Wistar rats were randomly divided into sepsis group and control group with 15 rats in each group. Cecal ligation and puncture (CLP) was used to reproduce rat sepsis model, and the success of reproduction was confirmed by examining the heart tissue with transmission electron microscope. Gene expression spectrum was studied with oligonucleotide gene expression profile microarray that contained 22 523 rat cDNA clones to detect the changes in gene expression pattern of rat heart tissue 24 hours after CLP. Genes with fluorescent signal of Cy3/Cy5 of ratio average (RA)>2.0 or RA<0.5 were identified as differential genes, then those that high correlated to sepsis were screened by means of related computer software, and their relationship was analyzed. Results Electron microscopic examination of heart tissue demonstrated that the sepsis model was successfully reproduced. Compared to the controls, gene expression of 418 genes of heart tissue of septic rat were changed 24 hours after CLP, accounting for 1.86%, and among them 200 genes showed up-regulation, 218 genes with down-regulation. Among known functional genes, 84 genes up-regulated and 74 genes down-regulated. They were related with a range of genetic functions, such as acute stress reaction, signal transduction, immune response, energy metabolism related genes etc. Conclusion There are a series of changes in gene expression in heart tissue apparently induced by sepsis rat. DNA microarray technology provides a new tool for rapidly analyzing them.%目的 应用基因芯片技术初步分析脓毒症大鼠心脏组织细胞基因表达谱的变化.方法 雄性Wistar大鼠30只被随机分为脓毒症组和对照组,每组15只.采用盲肠结扎穿孔术(CLP)制备大鼠脓毒症模型,以透射电镜下心脏组织检查鉴定模型.应用含有22 523个大鼠基因cDNA克隆的表达谱基因芯片进行检测,以Cy3和Cy5两种荧光信号强度结果 比值均>2.0或<0.5的基因为脓毒症表达差异基因,用计算机软件筛选并分析脓毒症大鼠心脏组织术后24 h的基因表达变化,并初步分析表达差异基因与脓毒症之间的关系.结果 与对照组比较,脓毒症组大鼠心脏组织共筛选出418个出现差异表达的基因,占基因芯片总点数的1.86%,其中表达上调者200个,表达下调者218个.在已知功能基因中表达上调者84个,下调者74个,与应激反应、细胞信号转导、糖皮质激素受体、免疫反应、胰岛素样生长因子、细胞物质能量代谢等方面的相关基因功能有关.结论 脓毒症大鼠心脏组织出现一系列基因表达异常,用基因芯片检测技术可快速分析.

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