骨形态发生蛋白和白血病抑制因子诱导肝干细胞分化  

Hepatic lineage differentiation of hepatic progenitor cells by bone morphogenetic protein or leukemia inhibitory factor

作　　者：高亚丽[1] 蔡雪飞[1] 刘娇[1] 单晓亮[1] 陈庆美[1] 周帆[1] 唐霓[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所感染性疾病分子生物学教育部重点实验室,400010

出　　处：《中华肝脏病杂志》2011年第9期692-695,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金（30972586）;重庆市自然科学基金重点项目（2009BA5036）;重庆医科大学校级重点项目（XBZD201011）

摘　　要：目的在永生化的小鼠肝干（祖）细胞（HP）模型上筛选并优化高效的肝细胞定向分化诱导方法，探讨HP向肝细胞定向分化过程及分子机制。方法分别采用含人白血病抑制因子（LIF）、骨形态发生蛋白（BMP）2和BMP9基因的重组腺病毒感染HP，在病毒感染后第4天、第7天和第10天用糖原染色和吲哚花青绿（ICG）摄取实验观察HP的分化成熟度，并在第4、7、10、14天通过检测白蛋白启动子调控的荧光素酶报告基因活性，观察细胞合成白蛋白情况。计量资料比较用t检验。结果BMP2和BMP9对HP的诱导作用最强，荧光素酶活性、PAS染色和ICG摄取细胞阳性率随诱导时间的延长明显上升，在诱导后第7天最高，HP对BMP9的诱导应答最强，与对照组相比，酶活性增加了近9倍（t=17．30，P〈0．01），BMP2处理组增加了5倍（f=16．41，P〈0．01），LIF处理组增加了3倍（t=6．04，P〈0．01）。诱导第7天时，PAS染色细胞阳性率在BMP2和BMP9组分别为30％和45％；ICG细胞阳性率在BMP2和BMP9组分别为40％和30%。LIF诱导后，PAS染色、ICG摄取细胞阳性率以及荧光素酶活性有一定增加。结论BMP2、BMP9和LIF能够诱导HP向发育晚期肝细胞分化，并初步具备成熟肝细胞的一些功能。Objective To search for the optimal approach for hepatocyte-directed differentiation of hepatic progenitor cells and investigate the molecular mechanism of the hepatic differentiation. Methods Hepatic progenitor cells were infected with recombinant adnovirus which containing human LIF, BMP2 or BMP9 gene. The maturation and differentiation of progenitor cells were examined by PAS sraining and ICG uptake methods at 4, 7 and 10 days post infection. The production of Albumin （Alb） was measured by luciferase activity at day 4, 7, 10 and 14. Results PAS staining assay revealed that BMP2 and BMP9 enhanced glycogen storage in hepatic progenitor cells most obviously at day 7. The percentages of positive cells were 30% and 45% respectively at 7 days post-infection. Meanwhile, 40% and 30% cells were positive by ICG uptake assay after BMP2 and BMP9 induction. Luciferase activity indicated that BMP9 induced ALBLuc activity most significantly at day 7. However, less inductive activity was found in LIF-treated group. Conclusion These results indicated tuat hepatic progenitor cells were differentiated into hepatocyte-like cells by BMPs and LIF induction.

关 键 词：干细胞 肝 细胞分化 骨形态发生蛋白质类 白血病抑制因子 永生化 

分 类 号：R363[医药卫生—病理学]