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.
出处
《中华肝脏病杂志》
CAS
CSCD
2003年第1期33-36,共4页
Chinese Journal of Hepatology
