摘要
目的阐明NDRG2(N-Myc downstream-regulated gene 2)在大鼠肝再生过程中的作用。方法大鼠70%肝切及除后收集0h、8 h、24 h、48 h、72 h、7 d、10 d的再生肝组织,采用Real-time PCR和Western blot方法检测大鼠肝再生过程中NDRG2及C-Myc基因和蛋白的动态变化。流式细胞仪检测腺病毒介导的高表达NDRG2对大鼠正常肝细胞系(BRL 3A)细胞周期和凋亡的影响。结果 NDRG2基因和蛋白的表达水平在肝再生达到高峰时明显下降,在肝细胞进入分化期时显著上调,其变化与C-Myc呈负相关;周期检测显示BRL3A细胞中高表达NDRG2 48 h后,G0/G1期细胞百分比从AdLacZ组的42.80±2.17上升至59.80±3.12,S期从35.91±1.15下降至12.09±2.21;凋亡检测显示NDRG2作用使细胞凋亡率较AdLacZ组升高5.9倍,差异有统计学意义(P<0.05)。结论 NDRG2通过影响细胞周期和凋亡参与调控大鼠肝再生过程。
Objective To demonstrate the role of N-Myc downstream-regulated gene 2（NDRG2） in rat liver regeneration.Methods The rats were subjected to 70% partial hepatectomy.Liver samples were collected at the indicated time points following surgery（0 h,8 h,24 h,48 h,72 h,7 d,10 d）.Real-time PCR and Western blot were performed to analyze the expressions of NDRG2 and C-Myc.BRL 3A cell lines were treated with AdNDRG2,AdLacZ or PBS for 48 h.Cell cycle and apoptosis were analyzed with a FACS Calibur flow cytometer.Results The levels of NDRG2 mRNA and protein strongly reduced when liver regeneration reached a peak of activity.In addition,it was found that rat NDRG2 expression was negatively correlated with C-Myc expression during this process.A low level of NDRG2 was observed as C-Myc expression increased during regeneration.Moreover,after been transfected with adenovirous,the proportions of cells in G_0/G_1 phase increased from 42.80±2.17 in the AdLacZ group to 59.80±3.12 in the AdNDRG2 group.Meanuhile,cells in S phase decreased from 35.91±1.15 to 12.09±2.21.In addition,the expression of NDRG2 dramatically increased cell apoptosis 5.9-fold compared with AdLacZ group.Conclusion NDRG2 regulated rat liver regeneration by its strong apoptosis-inducing activity as well as a cell cycle inhibitory effect.
出处
《胃肠病学和肝病学杂志》
CAS
2010年第5期440-443,共4页
Chinese Journal of Gastroenterology and Hepatology
