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Study on the apoptosis of rat hepatic stellate cells induced by kapamycin
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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。Abstract: 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.
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Key words:
- hepatic stellate cell /
- Sirolimus /
- apoptosis
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