骨髓基质干细胞对大鼠肝纤维化细胞凋亡的影响

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

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。

Objective To explore the effect on bone marrow stromal ceils （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. However, p75 was expressed in CFSCs, and the expression level increased after co-cuhure of CFSCs and BMSCs. The proportion of apoptotic 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.