摘要
目的分析热缺血损伤后大鼠肝脏基因表达谱的改变，初步确定热缺血损伤发病分子机制过程中调控基因。方法采用基因表达谱芯片RAE230分析SD大鼠原位热缺血模型处理前后基因mRNA转录本含量的改变；确定目标基因并联机检索GeneBank等整合数据库，按其生物功能分类进行批量筛选；并应用Gene Cluster3．0生物信息可视化分析软件进行等级聚类分析。用荧光定量逆转录聚合酶链反应法验证目标基因GAPDH、Hsp70、CyclinD1的表达变化，验证芯片分析的初步结果。结果分别得到了正常对照组、缺血组的肝脏基因表达数据和变化显著基因直观图示。对比分析发现，在总共8804个表达的基因（表达率为55．3％）中，实验组有927个基因表达下调，885个基因表达上调，其中变化幅度大于3倍的分别有40个和29个（P〈0．01）；按其生物学功能分类属能量代谢、组织修复、信号转导类分子的调控。结论30min热缺血损伤处理后，SD大鼠基因的表达谱发生改变，实验分析证实热缺血损伤的分子调控机制涉及多个基因表达调控途径。研究这些基因分子功能有助于为外科临床防治热缺血损伤提供实验依据。
Objectives To characterize a mRNA expression profile of unlethal warm ischemic injury in rat livers and to clarify the controlling genes and molecular pathogenesis. Methods One group of rats underwent surgeries to make a warm ischemic injury of their livers and another group of rats （controls） had sham operations; then 8 hybridized RNA samples were used in microarray analysis. The data were submitted to public databases, such as Gene Bank, dbEST, Unigene and Refseq, and then batch filtrated. Hierarchical clustering algorithm was applied in analyzing the differential expression genes and individual samples according to CeneCluster 3.0 software. Results Among the total 8804 probe sets, the expressions of 927 genes were downregulated and 885 genes were up-regulated in the iscbemia group when compared with normal controls. Between them, 40 genes were down-regulated and 29 genes up-regulated more than 3 fold （P 〈 0.01）; then Hsp70 and CyclinD1 were confirmed by real-time fluorescent quantitative RT-PCR test. Conclusion In the present study, we provide the first report of large-scale changes in hepatic gene expression induced by nonlethal warm ischemic injury. Most of up-regulated genes belong to the groups that are important in maintaining cell structure, division, differentiation, tricarboxylic acid cycle and tissue repair. Discovering such candidate genes of protective effects may be of help to prevent warm ischemic injury and to treat this serious disorder.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2007年第1期50-54,共5页
Chinese Journal of Hepatology
基金
广东省教育厅基金资助项目（Q02033）
China Medical Board in Newyork基金（06837）
关键词
肝
基因表达调控
热缺血损伤
微阵列
Liver
Gene expression regulation
Injury, ischemic
Microarray
