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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 investigate the effects of bone marrow stromal stem cells (BMSCs) on the apoptosis of rat hepatic fibrosis cells (CFSCs). Methods BMSCs, CFSCs and rat liver cell line BRL were cultured routinely, and co-cultured CFSCs and BMSCs. The concentrations of NGF, HGF and TGF-β1 in culture supernatant of BMSCs were detected by ELISA. The expression of p75 in CFSCs and BRL co-cultured with BMSCs was detected by RT-PCR. The apoptosis of CFSCs was detected by TUNEL method after secreted by BMSCs influences. Results BMSCs secreted cytokines such as NGF, HGF and TGF-β1, and secreted gradually with the prolongation of culture time. The expression levels of p75 and CFSCs were not significantly increased in BRL, p75 and CFSCs, Respectively with p75 blockers TAT-Pep5, HGF neutralizing antibodies and JNK blockers sp600125 role in BMSCs induced CFSCs apoptosis were significantly reduced; blocking TGF-β1, BMSCs induced CFSCs apoptosis ratio Increase. Conclusion BMSCs can promote the apoptosis of CFSCs through the secretion of NGF and HGF. The induction of apoptosis depends on the activity of JNK and is enhanced after the blocking of TGF-β1.