摘要
利用中效原理评价薯蓣皂苷元与肿瘤坏死因子相关凋亡诱导配体(TNF-related apoptosis-inducing li-gand,TRAIL)联合应用于非小细胞肺癌A549细胞的相互作用,并探讨可能的作用机制。MTT法观察两药对A549和人肾上皮293T细胞的增殖抑制作用。运用中效原理(Chou-Talalay合用指数法)判定两药的相互作用。Hoechst 33342荧光染色、流式细胞术检测两药联合对细胞凋亡的影响。Western blotting检测凋亡相关蛋白表达的变化。薯蓣皂苷元和TRAIL均能剂量依赖性地抑制A549细胞增殖。运用中效原理计算合用指数,当fa=0.1,CI>1;当fa>0.1,CI<1。对于A549细胞,薯蓣皂苷元与TRAIL合用,IC50由21.864μmol.L 1降低到14.810μmol.L 1(P<0.05);而对于293T细胞,合用前后IC50分别为27.129和27.184μmol.L 1(P>0.05)。荧光显微镜观察及流式细胞术检测结果表明,两药合用使细胞凋亡比例增加。联合用药组中Caspase-8、Caspase-9、Bid、Caspase-3激活和PARP切割明显增加;Bcl-2表达减少,而Bax表达增加;MAPK途径中p38、JNK和ERK明显激活。结果显示,薯蓣皂苷元和TRAIL具有协同诱导A549细胞凋亡的作用,机制与线粒体途径、死亡受体途径和MAPK信号途径均相关。
This study is to investigate the apoptotic induction effect of the combination of diosgenin and TNF-related apoptosis-inducing ligand（TRAIL） on non-small-cell lung cancer cell line A549 by using the ChouTalalay method,and observe the mechanism of the combination.The apoptotic effect of diosgenin or TRAIL alone and their combination on A549 and normal cell line 293T proliferation was measured by MTT assay.Chou-Talalay method was used to evaluate the combination effect.Apoptosis was examined by Hoechst 33342 staining and flow cytometry assay.Western blotting detects the expression of apoptosis-associated proteins.Diosgenin or TRAIL alone can inhibit proliferation of A549 in a concentration-dependent manner.According to the Chou-Talalay method,when fa= 0.1,CI 1,when fa 0.1,CI 1.Combined with TRAIL,the IC50 of diosgenin decreases from 21.864 to 14.810 μmol.L 1（P 0.05） on A549 cells.But for 293T cells,IC 50 of diosgenin does not change significantly.As with Hoechst 33342 staining and flow cytometry assay,the apoptosis ratios also increased in the combination group.At protein expression level,combination-treated group displays increased Caspase-8,Caspase-9,Bid,Caspase-3 activation and PARP cleavage,significantly decreased Bcl-2 and increased Bax expression,and MAPK pathways were activated.The combination of diosgenin and TRAIL has synergistic effect on A549 cells.
出处
《药学学报》
CAS
CSCD
北大核心
2013年第1期45-51,共7页
Acta Pharmaceutica Sinica
基金
澳门科技发展基金资助项目(071/2009/A3
091/2009/A)
