雷帕霉素诱导大鼠肝星状细胞凋亡的研究

目的观察雷帕霉素(RAPA)对大鼠肝星状细胞系rHSCs-99增殖、凋亡的影响。方法将rHSCs-99分成4组:对照组(A组),RAPA 50 nmol/L组(B组),RAPA 100 nmol/L组(C组),RAPA 150 nmol/L组(D组)。RAPA作用72 h后,分别用MTT法检测rHSCs-99增殖,ELISA法检测I型胶原表达,流式细胞仪Annexin-V FICT/PI法检测细胞凋亡情况,Wright-Giemsa染色法观察细胞形态学改变,细胞免疫组化法检测Fas、P53与Bcl-2的表达变化。结果 RAPA作用后,rHSCs-99增殖受到抑制,I型胶原含量降低,凋亡率增高,Fas、P53表达增多,Bcl-2表达减少。结论 RAPA能抑制rHSCs-99的增殖及I型胶原合成。促进rHSCs-99凋亡,其机制可能是通过开启Fas/FasL凋亡途径及上调凋亡相关基因P53、下调Bcl-2的表达来实现。

Study on the apoptosis of rat hepatic stellate cells induced by kapamycin

Objective To investigate the effect of Rapamycin(RAPA)on proliferation and apoptosis of rHSCs-99 in vitro.Methods The rHSCs-99 was divided into four groups-Control group(A),RAPA 50 nmol/L group(B),RAPA 100 nmol/L group(C),RAPA 150 nmol/L group(D).After 72 h of Rapamycin interference in rHSCs-99,the MTT colorimetric assay was used to detect the proliferation of rHSCs-99;synthesis and secretion of collagen I were detected by ELISA;Annexin-V FICT/PI double-label immuno fluorescence with flow cytometry was used to examine the apoptosis of rHSCs-99;Wright-Giemsa staining method was used to observe the morphologic change of rHSCs-99;Immunocytochemical staining was used to observe the proteinum expression of Fas,P53 and Bcl-2.Results After the rHSCs-99 were interfered with RAPA,rHSCs-99 proliferation were inhibited;the content of type I was reduced;the number of Fas,P53 positive was increased and the Bcl-2 positive was decreased.Conclusion RAPA inhibits the proliferation of the rHSCs-99 and reduces the content of collagen I of ones,facilitating the cell apoptosis.The mechanism which RAPA promote rHSCs-99 apoptosis may be through opening the Fas/FasL apoptosis pathway,up-regulating Fas、P53 and inhibiting Bcl-2 expressing to come true.