肝脏热缺血再灌注损伤对细胞周期调控相关基因表达的影响

The influence of warm ischemia-reperfusion injury on cell cycle control in liver

目的观察肝脏热缺血再灌注损伤(WIRI)对细胞周期调控相关基因表达的影响,探索 WIRI 的分子机制。方法将24只 SD 大鼠随机分为假手术组、热缺血30 min 组、再灌注1 h 组和24 h 组,利用基因芯片和实时荧光定量聚合酶链反应(real-time PCR)分析验证 WIRI 大鼠模型处理前后基因表达的动态变化;按基因表达的动力学特征对功能基因进行多元变量聚类分析。结果常温热缺血30 min、再灌注1 h 和24 h 时,肝实质细胞内 mRNA 表达上调的基因分别有86、410和234条,表达下调的基因有136、312和653条。经 real-time PCR 技术验证 Gadd45a、Egr1和 Hsp70的表达情况与芯片检测结果一致。结论采用非致死性 WIRI 处理大鼠后,再灌注早期肝脏细胞周期调控基因和再生相关基因表达上调,再灌注24 h 后多数早期上调基因恢复到 WIRI 处理前水平,证明 WIRI 早期即通过细胞周期调控基因影响细胞周期。Gadd45a、Egr1和 Hsp70基因有可能成为药物治疗 WIRI的新靶点。

Objective To observe the effect of warm ischemia-reperfusion injury （WIRI） on the genes involved in the cell cycle in liver and investigate the molecular mechanism of WIRL Methods Twenty- four Sprague-Dawley male rats were divided into experiment groups and sham operation group respectively. According to the dynamic characteristics, the genes involved in the cell cycle were submitted for multivariate cluster and self-organized map analysis. Total RNA was extracted for the microarray analysis and subsequent real-time quantitative PCR. Results There were 86,410,234 up-regulated genes and 136,312,653 downregulated genes over 2-4 times in the 0 h ,1 h and 24 h group respectively,most of which such as Gadd45a, Egr1 ,Hsp70 were involved in the cell cycle checkpoint, energy metabolism, signal transduction, immune- associated procedure,and so on. In addition, the results of microarray analysis were comfirmed by real-time quantitative PCR. Conclusions Some genes related with cell cycle and regeneration were activated in the early stage of WIRL Gadd45a,Egr1 and Hsp70 may be new targets to cure WIRI.