骨髓基质干细胞对大鼠肝纤维化细胞凋亡的影响

目的探讨骨髓基质干细胞(BMSCs)对大鼠肝纤维化细胞(CFSCs)凋亡的影响。方法常规培养BMSCs、CFSCs和大鼠肝细胞系BRL,并双层共培养CFSCs与BMSCs。用ELISA方法检测BMSCs培养上清中NGF、HGF和TGF-β1的浓度;RT-PCR检测与BMSCs共培养的CFSCs及BRL的p75表达;TUNEL法检测BMSCs分泌的细胞因子封闭后对CFSCs凋亡的影响。结果BMSCs可分泌NGF、HGF、TGF-β1等细胞因子,且随着培养时间的延长,其分泌量逐渐增多;BRL不表达p75,CFSCs表达p75,且与BMSCs共培养后,其表达量增高;分别用p75的阻断剂TAT-Pep5、HGF的中和抗体和JNK的阻滞剂sp600125作用后,BMSCs诱导CFSCs凋亡的比例均明显降低;封闭TGF-β1后,BMSCs诱导CFSCs凋亡的比例增高。结论BMSCs能够通过分泌NGF和HGF促进CFSCs的凋亡,这种凋亡诱导作用依赖于JNK的活性,并且在封闭TGF-β1后增强。

Effect of Bone Marrow Stromal Cells on Apoptosis of Rat Cirrhotic Fat-storing Cells

Objective To explore the effect on bone marrow stromal cells(BMSCs)on the apoptosis of rat cirrhotic fat-storing cells(CFSCs).Methods BMSCs,CFSCs and rat liver BRL cells were cultured by routine methods,and CFSCs and BMSCs were subjected to two-layer co-culture.Determine the concentrations of NGF,HGF and TGF-β1 in culture supernatant of BMSCs by ELISA,the expression of p75 in CFSCs after co-culture with BMSCs as well as in BRL cells by RT-PCR,and the effect of cytokines secreted by BMSCs,after blocking,on the apoptosis of CFSCs by TUNEL method.Results BMSCs secreted cytokines such as NGF,HGF and TGF-β1,and the secretion level increased with the increasing time for culture.No p75 was expressed in BRL cells.How-ever,p75 was expressed in CFSCs,and the expression level increased after co-culture of CFSCs and BMSCs.The proportion of apop-totic CFSCs induced by BMSCs after treatment with the blocking agent TAT-Pep5 of p75,the neutralizing antibody against HGF and the paralyser sp600125 of JNK decreased significantly.However,after blocking of TGF-β1,the proportion increased.Conclusion BMSCs promoted the apoptosis of CFSCs by secreting NGF and HGF.The promoting effect was JNK activity-dependent and increased after blocking of TGF-β1.