摘要
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。
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 1130 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 EL1SA ; 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.
出处
《临床肝胆病杂志》
CAS
2012年第3期219-222,共4页
Journal of Clinical Hepatology
