细胞生长因子体外对大鼠肝干细胞的影响  

Mitogenic effects of growth and differentiation factors on rat liver stem cell WB-F344 in vitro

作　　者：姚鹏[1] 胡大荣[1] 詹轶群[2] 许望翔[2] 李长燕[2] 杨晓明[2] 

机构地区：[1]北京军区总医院肝病研究所,北京100700 [2]军事医学科学院放射研究所

出　　处：《中华肝脏病杂志》2003年第1期33-36,共4页Chinese Journal of Hepatology

摘　　要：目的研究大鼠肝干细胞系WB-F344细胞在体外的生长分化与多种细胞因子的关系.方法用3H掺入法检测细胞因子对WB-F344细胞生长增殖作用.用western blot检测WB-F344细胞因子受体的表达.流式细胞仪检测细胞凋亡.结果发现肝细胞生长因子(HGF)80ng/ml cpm值为982.95、表皮生长因子(EGF)80 ng/ml cpm值为906.32、胰岛素80 ng/ml cpm值为968.67、成纤维细胞生长因子(FGFα)80 ng/ml cpm值为863.98,促进WBF344细胞的增殖;白细胞介素-6(IL-6)80 ng/ml cpm值为368.67、肿瘤坏死因子α 80 ng/ml cpm值为372.90对WB-F344细胞的生长无明显影响;而转化生长因子(TGF β)能够诱导WB-F344细胞凋亡(80 ng/ml凋亡率为26.89%)并抑制其生长;western blot检测发现WB-F344细胞表面存在HGF、EGF、FGF α、TGF β等细胞因子受体.提示细胞因子可能通过与其受体结合方式调控WB-F344细胞的生长分化.进一步用HGF、EGF、胰岛素等细胞因子组成并添加地塞米松的分化培养体系,在体外对WB-F344细胞进行定向诱导分化.发现WB-F344细胞在体外能够向肝实质细胞分化.此外HGF还具有刺激肝干细胞离散作用.结论肝干细胞的生长分化受多种细胞因子的调控,细胞生长因子可能在严重肝损伤后肝干细胞的动员、增殖及分化调控中起重要作用.Objective To explore the relationship between the proliferation, differentiation of rat hepatic stem like cell line WB-F344 and cytokines in vitro. Methods 5H thymidine labelling of new synthesized DNA was used to examine the mitogenic responsiveness of WB-F344 cells to cytokines, western blot was used to study the expression of cytokines receptors on hepatic stem cells, and apoptotic cells were detected by Flow cytometry. Results WB-F344 cells showed a proliferative response to the cytokines of hepatocyte growth factor (HGF), epidermal growth factor (EOF), fibroblast growth factor (FGF), Insulin at the dose of 80 ng/ml, and the relative cpm values are 982.95, 906.32, 863.98 and 968.67 respectively, while non response to interleukin-6 (IL-6), tumor necrosis factor-a (TNFa ) at the same dose, and an inhibition or apoptosis response to transforming growth factor-(5 (TGF (3) at 80 ng/ml with a 26.89% apoptotic rate. Western blot showed that there were HGF, EGF, FGF, TGF (5 receptors expressed on WB-F344 cells. When WB-F344 cells were cultured in the differential system (DMEM, 10% Fes, HGF 10-50 ng/ml, EGF 20 ng/ml, Insulin 1 ug/rnl, Dex 1 umol/L), the cells could differentiated into hepatocytes. In addition, HGF could scattered WB-F344 cells. Conclusions The proliferation and differentiation of liver stem cells are regulated by various cytokines which may play an important role when liver is damaged seriously.

关 键 词：肝损伤 分子生物学 表皮生长因子 细胞生长因子 大鼠 肝干细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]