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目的肝脏卵圆细胞(HOC)在二甲基亚硝胺(DMN)致大鼠肝硬化形成与消减过程中表达的动态变化,探讨其病理生理意义。方法应用DMN腹腔注射(4周12次)制备大鼠肝硬化模型,分别于造模后3d、2、4周及终止造模后2,4周处死大鼠取肝组织,进行常规组织学观察,透射电镜下观察HOC 超微结构,免疫组织化学和western blot方法检测Thy1.1的表达变化,应用图像分析法对所标记的HOC进行半定量测定以及光镜下计数细胞数量。结果DMN造模4周大鼠已形成典型的肝硬化;终止造模后2周时肝组织病变较造模4周时有所减轻,终止造模后4周时炎症明显减轻并以不完全纤维间隔为主。Thy1.1 阳性染色细胞正常大鼠肝组织未见到,造模后3 d时可见微量表达,2周时呈散在分布,4周时明显增多,见于纤维间隔周围,终止造模后2周、即第6周时阳性染色显著强于造模4周,大量出现于汇管区,8周时较6周有所减少,表达量基本与4周时相等。阳性染色图像分析、光镜下阳染细胞计数及western blot三者的结果呈一致性。Western blot结果与免疫组织化学观察结果也具一致性。电镜下显示HOC具有形态小,核以卵圆型为主,高的核/浆比例等特点。结论Thy1.1阳性的HOC可能与肝硬化的消减或逆转有关。
Objective To investigate the dynamic changes of hepatic oval cell (HOC) expression during the formation and abatement of cirrhosis caused by dimethylnitrosamine (DMN) in rats and its pathophysiological significance. Methods The rat model of hepatic cirrhosis was established by intraperitoneal injection of DMN (12 times a week for 12 weeks). The rats were sacrificed on the 3rd, 4th, 4th week and the 2nd and 4th week after the model was established, respectively. The ultrastructure of HOC was observed under transmission electron microscope. The expression of Thy1.1 was detected by immunohistochemistry and western blot. Semi-quantitative determination of the labeled HOC and count of cells under light microscope were performed by image analysis. Results Four weeks after modeling, DMN formed typical liver cirrhosis. At 2 weeks after the model was established, the pathological changes of liver tissue were lessened 4 weeks after model establishment. Inflammation was relieved 4 weeks after the model was established. Interval-based. Thy1.1 positive staining cells in normal rat liver tissue not seen, 3 d after modeling can be seen micro-expression, scattered distribution at 2 weeks, 4 weeks was significantly increased, seen around the fiber interval, 2 weeks after the termination of modeling, The positive staining was significantly stronger at the 6th week than that of the model 4 weeks, and appeared in the portal area at the 8th week. The expression level was basically the same as that at 4 weeks. Positive staining image analysis, counting under the light microscope and western blot results of the three were consistent. Western blot results and immunohistochemical findings are also consistent. Electron microscopy showed that HOC has the characteristics of small shape, mainly oval-shaped nucleus and high nuclear / plasma ratio. Conclusion Thy1.1-positive HOC may be related to the reduction or reversal of cirrhosis.