骨髓间充质干细胞在肝癌前病变大鼠肝脏内的分化及槲芪散的影响

The differentiation of the bone marrow-derived liver stem cells in rat livers with precancerous lesion and the role of Huqi San

目的:探讨骨髓源性肝脏干细胞是否参与肝癌前病变的形成,评价槲芪散对肝癌前病变的干预作用。方法:取80g体重的雄性Wistar大鼠的股骨和胫骨,用PBS冲洗骨髓,按密度梯度离心法原代分离培养骨髓间充质干细胞(mesenchymal stemcell,MSCs),用绿色荧光蛋白(GFP)作为追踪标记物将MSCs进行标记。将大鼠分为6组:即常规饲养组、模型组、模型+MSCs移植组、肝大部切除+MSCs移植组、槲芪散治疗组8g/kg、槲寄生总碱治疗组8g/kg。除正常对照组外,各组动物在行肝大部切除术后,选择剩余肝脏不同部位表面注射经GFP标记好的MSCs,用荧光显微镜观察GFP标记MSCs并计算标记率。用免疫荧光化学的方法检测肝脏内GFP标记的细胞。用组织化学染色的方法及Western Blot的方法分别检测大鼠肝脏-GT的表达及大鼠肝脏AFP的表达。结果:模型组及及模型+MSCs移植组的肝脏中-GT灶的数量及面积显著增多,且AFP的表达明显升高,以模型+MSCs移植组的变化更为明显,与槲芪散及槲寄生碱组比较有显著性差异。结论:MSCs参与了肝癌前病变的发生发展过程,而槲芪散及槲寄生总碱能够改善肝脏内环境,影响归巢于肝脏的MSCs的分化方向,诱导MSCs分化为正常肝细胞。

To study the involvement of mesenchymal stem cells with precancerous lesion ; And to investigate the effect of HQS in intervention the occurrence and development of precancerous lesion. Methods： MSCs were extracted from 80g-weight male Wistar rats for primary culture. The MSCs were marked with GFP. There were 6 groups, including normal group, model group, model transplantation group, partial hepatectomy and MSCs transplantation group, HQS treatment group, MA treatment group. We chose different parts of left liver to inject GFP marked MSCs except the normal group. GFP marked MSCs were determined by immunofluoreseenee. -Glutamy-transpeptidase-isoenzyme was determined with histochemical methods. Alpha-Fetoprotein was determined with Western Blot methods. Results ： The areas of -GT positive fo- ci were i^creased in model group and model transplantation group. AFP of transplantation group was significantly increased compared with HQS treatment group and MA treatment group （ P 0.05）. Conclusion： MSCs might be involved in the formation of hepatic precancerous lesion. HQS and MA can improve the enviroment of liver and induce MSCs to differentiate normal hepatocyte.