热缺血再灌注损伤影响肝脏细胞周期检查点调控的基因表达分析

目的动态观察肝脏热缺血再灌注损伤（WIRI）对细胞周期检查点基因的表达影响，研究其分子机制帮助了解那些与热缺血再灌注相关的疾病。方法采用成组对照的实验设计，利用微阵列芯片和逆转录-聚合酶链反应（PCR）联合分析技术，分析SD大鼠热缺血再灌注损伤处理后的基因表达变化（其中芯片分析8只、RT-PCR定量分析16只）；运用系统生物学分析方法对功能基因进行聚类。结果分别得到了热缺血再灌注损伤处理后肝脏的细胞周期各检查点基因及早期即刻基因表达数据，将其中与细胞周期密切相关的的功能基因（上调的223条、下调的62条变化幅度均大于3倍）做进一步分析。结论肝脏热缺血再灌注损伤通过细胞周期／检查点控制基因影响细胞周期G1／S、G2／M的运行，抑制DNA的合成和染色体的分裂。推测可能影响受损的DNA双链自我修复和后期的组织细胞再生、凋亡。

Genes expression analysis of cell cycle control in liver warm ischemia-reperfusion injury with microarrays

Objective To observe the effect of warm ischemia-reperfusion injury (WIRI) on the cell cycle/checkpoint gene expression in liver and investigate the molecular mechanism. Methods Four groups of SD rats underwent WIRI and sham operation respectively (8 samples analyzed by RAE230-chip and 16 by RT-PCR respectively). With reference to the Public Gene database,the target genes were analyzed according to their biophysiological functions. Results The expression data of each group were obtained, and it was found that the checkpoint genes which arrested in G1/S and G2/M phase were changed significantly (223 genes up-regulated and 62 genes down-regulated more than 3 times). The following quantitative RT-PCR tests were in coincidence with the chip results. Conclusion The ischemia-reperfusion injury has effects on the checkpoint genes,unregulates the cell cycle G1/S,G2/M arrest,and prompts the GO transfer into the G1 phase at the same time. Moreover,these active genes may manipulate the self-repair of impaired DNA,cell differentiation and apoptosis.