肝脏卵圆细胞定向分化在二甲基亚硝胺大鼠肝硬化消减过程中的意义

Significance of oriented differentiation of hepatic oval cells in course of formation and reduction of rat cirrhosis treated with dimethylnitrosamine

[目的]探讨肝脏卵圆细胞在肝硬化形成及修复过程中的动态变化及其意义。[方法]二甲基亚硝胺10 mg.kg-1剂量大鼠腹腔注射,每周连续3 d,1次/d,共4周,制备肝硬化模型;于二甲基亚硝胺腹腔注射后3 d、2周、4周及终止造模刺激后2周、4周分别随机取造模大鼠6只、正常大鼠3只进行动态观察。透射电镜下观察肝脏卵圆细胞超微结构,免疫组织化学和免疫印迹方法检测肝组织α-SMA及Thy1.1;双重荧光免疫共聚焦激光扫描显微镜技术观测肝组织AFP与Thy1.1及CK19与Thy1.1共定位。[结果]模型动态变化:大鼠经DMN造模4周时形成典型肝硬化病理改变;终止造模刺激2周后,肝组织学变化明显减轻。AFP、CK19与Thy1.1共定位细胞:造模3 d明显表达;随着造模时间延长表达量逐渐增加;造模停止后2周达到峰值,显著高于造模4周(P<0.05);而造模停止后4周显著少于造模停止后2周时的水平(P<0.01),但仍高于造模2周。[结论]肝脏卵圆细胞增殖与分化在DMN所致大鼠肝硬化的形成与消减过程中、尤其对促进硬化肝脏的组织重构具有重要作用。

[ Objective] Study the of the significance dynamic changes of hepatic oval cells in the course of formation and reduction of rat cirrhosis treated with dimethylnitrosamine. [ Methods] Rat cirrhosis models were established with dimethylnitrosamine injection by abdominal cavity, once a day, continuous 3 days a week, 4 weeks altogether. On the third day, second week, forth week of injection, and second week and forth week after stopping injection, 6 modeling and 3 normal rats were sacrificed respectively for dynamical observation. Uhrastructure of hepatic oval cells were detected by transmission electron microscope, expression of Thyl. land α - SMA by immunohistochemistry and immuoblot, collocation of AFP, CK19 and Thyl. 1 by double immuno confocal laser scan microscopy. [ Resuits ] There were dynamic changes of hepatic oval cell and the regulatory factors in the course of formation and reduction of rat cirrhosis treated with dimethylnitrosamine： There was typical cirrhosis in the liver on the forth week after the rats were treated with dimethylnitrosamine, the cirrhosis relieved notably on the second week after stopping dimethylnitrosamine. Collocation of AFP, CK19 and Thy1. 1 in the liver： There was notable expression of collocation on the third day of modeling; with the time being, the expression increased, which attached to the peak on the second week after stopping dimethylnitrosamine, more than that on the forth week of modeling（ P 〈 O. 05 ）, which dropped on the forth week after stopping dimethylnitrosaminc less than that on the second week after stopping dimethylnitrosamine （ P 〈 0.01 ）, but still higher than that on the second week of modeling. [Conclusion ] There is a great significance in reproduction and oriented differentiation of hepatic oval cell in the course of formation and reduction of rat cirrhosis especially in promoting reconstruction of liver tissue which encounter cirrhosis.