中药复方脏毒清体外诱导人结肠癌SW480细胞凋亡的影响  

Effect on Apoptosis of Human Colon Cancer SW480 Cells by Traditional Chinese Medicine Compound of Zangduqing

作　　者：安永康[1] 荫晴[2] 张双喜[2] 韩海涛[3] 吴芳[3] 刘晓博[3] 张相安[3] 

机构地区：[1]河南中医学院,郑州450000 [2]河南省中医药研究院附属医院,郑州450000 [3]河南中医学院第一附属医院,郑州450000

出　　处：《中国实验方剂学杂志》2015年第7期155-158,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省中医药科学研究专项(2013ZY03017)

摘　　要：目的:探讨中药复方脏毒清对人结肠癌SW480细胞的促凋亡作用及机制。方法:体外培养人结肠癌细胞株SW480,用不同质量浓度的脏毒清(0.05,0.10,0.15,0.20,0.25 g·m L-1)进行干预12,24,36,48 h后,采用MTT法检测细胞增殖抑制率。用不同质量浓度脏毒清(0.10,0.15,0.20 g·m L-1)作用SW480细胞24 h后,另设空白组,采用用荧光染色技术和流式细胞仪检测细胞凋亡率,分光光度法测定半胱天冬酶-3(Caspase-3),Caspase-9的酶活性,Western blot技术检测凋亡蛋白Bax,Bcl-2蛋白的表达情况。结果:MTT结果显示脏毒清对其有明显的增殖抑制作用,并呈现时间和剂量依赖性;与空白组比较,脏毒清0.10,0.15,0.20 g·m L-1组能明显诱导SW480细胞凋亡,Caspase-3及Caspase-9酶活性明显升高,促凋亡蛋白Bax表达明显升高,抗凋亡蛋白Bal-2表达明显降低,均具有统计学意义(P<0.01)。结论:脏毒清具有促进人结肠癌SW480细胞凋亡的作用,并通过线粒体凋亡途径发挥作用。Objective： To investigate the inducing apoptosis effect and the underlying mechanism of Zangduqing （ZDQ） on human colon cancer SW480 cells. Method： SW480 ceils were cultured with different concentrations of ZDQ （0.05, 0. 10, 0.15, 0.20, 0.25 g·mL^-1） for 12, 24, 36, 48 h in vitro. The cellular cytotoxicity was measured by using the MTT method. Then, SW480 cells were cultured with ZDQ （0.10, 0. 15, 0.20 g·mL^-1 ） for 24 h in vitro and a control group was assigned. The apoptosis was detected by the DAPI and annexin V-FITC/propidium iodide assay. The activities of Caspase-3, -9 were assayed by using Caspase apoptosis detection kit. Protein levels of Bax and Bcl-2 were determined by using Western blot. Result： MTT showed that ZDQ had obvious inhibiting effect on the proliferation of human colon cancer SW480 cells with a dose- and time- dependent manner. Compared with the control group, the ZDQ could significantly inhibit the proliferation of SW480 ceils, enhance the activity of Caspase-9 and Caspase-3, up-regulate the level of Bax, down-regulate the level of Bcl-2 （P 〈 0.01 ）. Conclusion： ZDQ could induce the apoptosis of SW480 cells obviously, which may be mediated by the mitochondrial apoptosis pathway.

关 键 词：脏毒清 结肠癌 SW480细胞 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]