摘要
目的:观察扶正抗癌方对H1650细胞凋亡的影响,并探讨其诱导H1650细胞凋亡的分子机制。方法:以人肺腺癌细胞株H1650细胞为研究对象,用0.5,1.0,1.5,2.0,2.5,3.0 g·L^(-1)扶正抗癌方处理H1650细胞24,48,72 h,另设空白组,四氮唑蓝盐化合物(MTS)法检测细胞增殖;用0.5,1.0,1.5 g·L^(-1)扶正抗癌方处理H1650细胞24 h,另设空白组,Annexin VFITC/碘化丙啶(PI)流式细胞术检测细胞凋亡;用0.5,1.0,2.0 g·L^(-1)扶正抗癌方处理H1650细胞24 h,另设空白组,半胱氨酸蛋白酶-3/7(Caspase-3/7)活力检测试剂盒检测Caspase-3/7活力;蛋白免疫印迹法检测扶正抗癌方对pro Casapse-3,聚腺苷二磷酸-核糖聚合酶(PARP),Bcl-2相关X蛋白(Bax)表达。结果:与空白组比较,扶正抗癌方能明显抑制H1650细胞的增殖(P<0.05),且呈浓度和时间依赖性。与空白组比较,扶正抗癌方明显诱导H1650细胞的早期凋亡,增强Caspase-3/7的活力(P<0.05),且呈浓度依赖性。与空白组比较,扶正抗癌方明显下调pro Casapse-3和PARP的表达(P<0.05),呈浓度依赖性,明显上调Bax的表达(P<0.05),且呈时间依赖性。结论:扶正抗癌方可通过激活Caspase-3和Bax诱导H1650细胞的凋亡。
Objective： To observe the effects of Fuzheng Kang＇ai （FZKA） decoction on apoptosis of H1650 cells and discuss its molecular mechanism in apoptosis of H1650 cells. Method： Human lung adenocarcinoma cells （H1650 cells） were used as the research objects and treated for 24, 48,72 h with 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 g·L-1FZKA decoction, and a blank group was set up, then cell proliferation was detected by 3- （4, 5-diethylthiazol-2-yl ） -5- （ 3-carboxymethoxyphenyl ） -2- （ 4-sulfophenyl ） -2 H-etrazolium, inner salt （ MTS ） assay. H1650 cells were treated for 24 h with 0.5, 1.0, 1.5 g·L-1 FZKA decoction and a blank group was set up, then cell apoptosis was detected by Annexin V-FITC/PI flow cytometry analysis. H1650 cells were treated for 24 h with 0.5, 1.0, 2.0 g·L-1 FZKA decoction and a blank group was set up, then vitality of Caspase-3/7 was detected by Caspase-3/7 vitality test kit. Simultaneously, effects of FZKA decoction on the expression levels of proCasapse- 3, PARP and Bax were detected by Western blot assay. Result ： As compared with the blank group, proliferation of H1650 ceils was significantly inhibited by FZKA decoction in concentration-dependent and time-dependent manners （P 〈 0.05）. As compared with the blank group, early apoptosis of H1650 cells was significantly induced and vitality of Caspase-3/7 was increased by FZKA decoction in a concentration-dependent manner （P 〈 O. 05）. As compared with the blank group, the expression levels of proCasapse-3 and PARP were significantly reduced by FZKA decoction in a concentration-dependent manner （P 〈 0. 05） , and the expression of Bax was increased by FZKA decoction in a time-dependent manner （P 〈0. 05）. Conclusion ： Apoptosis is induced by FZKA decoction in way of activating Caspase-3 and Bax in H1650 ceils.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第14期106-110,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81273965
81503507)
广东省自然科学基金项目(2015A030310245)
广东省建设中医药强省科研项目(20141104)
