摘要
目的了解大鼠肝外胆管缺血再灌注损伤后成纤维细胞的凋亡和增殖情况及其相关基因表达的时序性变化。方法检测缺血大鼠肝外胆管再灌注后1、3、24、72 h肝外胆管壁成纤维细胞的凋亡指数(TUNEL标记法)、Fas和增殖细胞核抗原(PCNA)蛋白的表达状况(免疫组织化学法)以及Bcl-2 mRNA的表达状况(原位杂交法)。结果缺血再灌注后1 h,成纤维细胞的凋亡指数以及Fas、PCNA和Bcl-2 mRNA表达与对照组无明显差异。再灌注后3 h,成纤维细胞凋亡指数升高,24 h时达到高峰,此后逐渐下降。Fas蛋白表达与凋亡同步;PCNA表达在再灌注后3 h时已明显增强,接近峰值水平,此后一直维持较高水平。再灌注3 h时Bcl-2 mRNA表达与对照组无明显差异,24 h迅速升高并达到峰值,此后逐渐下降。结论肝外胆管缺血再灌注早期成纤维细胞有一定程度的凋亡,但随后减弱,其增殖在早期即明显加强,且一直保持到再灌注后期。
Objective To investigate the temporal changes of fibroblast apoptosis and gene expression during extrahepatic bile duct ischemia/reperfusion. Methods At 1,3,24,48 and 72 h after 30 min＇ s extrahepatic bile duct ischemia in 50 rats, apoptotic index of fibroblasts was calculated by TUNEL staining, the expression of bcl-2 mRNA was evaluated by in shu hybridization and Fas and proliferating cell nuclear antigen （PCNA） expression were evaluated by immunohistochemistry. Results At the first hour after ischemia/reperfusion, no significant changes were observed in the expression of Fas, PCNA and Bcl-2 mRNA between the ischemic and the control groups. The increase of apoptotic index of fibroblast was first seen at 3h after reperfusion, and up to peak at 24 h and declined gradually thereafter. The expression of Fas concurred with the apoptotic index. The expression of PCNA appeared at 3 h after reperfusion and remained high from 24 h to 72 h. The expression of Bcl-2 mRNA peaked at 24 h after reperfusion. Conclusion The extrahepatic bile duct ischemia/reperfusion resulted in both apoptosis and proliferation of fibroblast, and apoptosis only appeared at early phase but proliferation maintained to the end of the experiment.
出处
《解剖学报》
CAS
CSCD
北大核心
2005年第6期625-628,共4页
Acta Anatomica Sinica
基金
中国宝山钢铁集团上海梅山公司科研基金(03-科-42)
