CAJ | 학술논문

目的观察哺乳动物雷帕霉素靶蛋白(mTOR)信号通路在Fas配体(Fas-L)诱导胆管细胞凋亡中的作用.方法以正常孵育胆管细胞为对照,Fas-L(1.5 μg/L)孵育原代培养2～10代大鼠胆管细胞48 h,流式细胞技术检测Fas-L对胆管细胞的凋亡诱导作用,Western blot检测两组胆管细胞内mTOR/p70S6K磷酸化水平,信号通路抑制剂雷帕霉素(Rapamycin,10 nmol/L)孵育两组胆管细胞24 h,观察胆管细胞内mTOR/p70S6K磷酸化水平.结果 Fas-L可显著诱导胆管细胞凋亡[(68.5±8.8)％比(6.9±1.2)％,P＜0.05],Fas-L孵育胆管细胞mTOR/p70S6K磷酸化水平升高,信号通路活化.雷帕霉素可显著抑制mTOR信号通路的磷酸化活化,明显减少Fas-L诱导的胆管细胞凋亡[(65.8±9.1)％比(20.9±4.3)％,P＜0.05].结论 Fas-L可能通过活化mTOR信号通路诱导胆管细胞凋亡.
목적 관찰포유동물뢰파매소파단백(mTOR)신호통로재Fas배체(Fas-L)유도담관세포조망중적작용.방법 이정상부육담관세포위대조,Fas-L(1.5 μg/L)부육원대배양2～10대대서담관세포48 h,류식세포기술검측Fas-L대담관세포적조망유도작용,Western blot검측량조담관세포내mTOR/p70S6K린산화수평,신호통로억제제뢰파매소(Rapamycin,10 nmol/L)부육량조담관세포24 h,관찰담관세포내mTOR/p70S6K린산화수평.결과 Fas-L가현저유도담관세포조망[(68.5±8.8)％비(6.9±1.2)％,P＜0.05],Fas-L부육담관세포mTOR/p70S6K린산화수평승고,신호통로활화.뢰파매소가현저억제mTOR신호통로적린산화활화,명현감소Fas-L유도적담관세포조망[(65.8±9.1)％비(20.9±4.3)％,P＜0.05].결론 Fas-L가능통과활화mTOR신호통로유도담관세포조망.
Objective To explore the roles of mammalian target of rapamycin (mTOR) signaling pathway in Fas-ligand (Fas-L)-induced cholangiocyte apoptosis.Methods Control study with primarily cultured rat cholangiocytes served as control group.Primarily cultured cholangiocytes were treated with Fas-L (1.5 μg/L) for 48 h.The flow cytometry was used to detect the apoptosis of cholangiocytes in response to Fas-L.Meanwhile, the activation of mTOR signaling pathway in cholangiocytes in response to Fas-L was detected by Western blotting.The cholangiocytes incubated with Fas-L were further treated by the specific inhibitor of mTOR signaling pathway, rapamycin (10 nmol/L) for 24 h to observe its influence on activation of mTOR signaling pathway and apoptosis.Results Fas-L significantly induced apoptosis of cholangiocytes [(68.5 ± 8.8) ％ vs.(6.9 ± 1.2) ％, P ＜ 0.05] and activated the mTOR signaling pathway by phosphorylating the mTOR and p70S6K.However, rapamycin not only significantly inhibited phosphorylation of mTOR/p70S6K but also reduced apoptosis of cholangiocytes induced by Fas-L [(65.8 ± 9.1) ％ vs.(20.9 ± 4.3) ％, P ＜ 0.05].Conclusion Fas-L induces apoptosis of cholangiocytes probably through activating mTOR signaling pathway.