伪士的宁对人结肠癌HT-29细胞凋亡的影响及其机制研究  

Study on the Effect and Its Mechanism of Pseudostrychnine on Human Colon Cancer HT-29 Cells

作　　者：冯振宇[1] 孟霜[1] 常剑敏 周晓荣 史敏 赵建平[1] FENG Zhenyu;MENG Shuang;CHANG Jianmin;ZHOU Xiaorong;SHI Min;ZHAO Jianping(Central Laboratory,Shanxi Hospital of Integrated Traditional and Western Medicine,Taiyuan 030013,China;College of TCM,Shanxi University of TCM,Taiyuan 030024,China)

机构地区：[1]山西省中西医结合医院/山西中医学院中西医结合医院中心实验室,太原030013 [2]山西中医药大学中药学院,太原030024

出　　处：《中国药房》2020年第2期179-183,共5页China Pharmacy

基　　金：国家国际科技合作专项项目(No.2014DFA33150)

摘　　要：目的:探讨伪士的宁对人结肠癌HT-29细胞凋亡的影响及其机制。方法:取人结肠癌HT-29细胞,随机分为空白组和低、中、高剂量伪士的宁组(125、250、500μmol/L),加入不含药培养基或含相应浓度伪士的宁的培养基培养48 h。采用流式细胞仪检测细胞凋亡情况和线粒体跨膜电位;采用Western blotting法检测细胞中P53、Caspase-3、Caspase-9、兔源DNA修复酶(c-PARP)、Bcl-2的蛋白表达水平。结果:与空白组比较,低、中、高剂量伪士的宁组细胞凋亡率显著升高,线粒体跨膜电位均显著降低(P<0.01),且均呈现浓度依赖趋势。中、高剂量伪士的宁组细胞中P53、Caspase-3、Caspase-9、c-PARP蛋白表达水平较空白组均显著升高,低、中、高剂量伪士的宁组细胞中Bcl-2蛋白表达水平则显著降低(P<0.05或P<0.01)。结论:伪士的宁可能通过上调P53蛋白、下调Bcl-2蛋白的表达,改变线粒体膜电位,然后激活Caspase-3、c-PARP、Caspase-9的表达,进而激活内源性线粒体通路,发挥促进HT-29细胞凋亡的作用。OBJECTIVE:To investigate the effects and its mechanism of pseudostrychnine on the apoptosis of human colon cancer HT-29 cells. METHODS:Human colon cancer HT-29 cells were randomly divided into blank group,pseudostrychnine lowdose,medium-dose and high-dose groups(125,250,500 μmol/L). They were cultured with culture medium without medicine or relevant concentration of pseudostrychnine for 48 h. The cell apoptosis and mitochondrial transmembrane potential were detected by flow cytometry. Western blotting assay was employed to detect the protein expression of P53,Caspase-3,Caspase-9,DNA repair enzyme from rabbit(c-PARP)and Bcl-2. RESULTS:Compared with blank group,apoptotic rate of cells was increased significantly in pseudostrychnine low-dose,medium-dose and high-dose groups,while mitochondrial transmembrane potential was decreased significantly(P<0.01),in concentration-dependent manner. The protein expression levels of P53,Caspase-3,Caspase-9 and cPARP were increased in pseudostrychnine medium-dose and high-dose groups,compared with blank group;while those of Bcl-2 were decreased significantly in pseudostrychnine low-dose,medium-dose and high-dose groups(P<0.05 or P<0.01). CONCLUSIONS:Pseudostrychnine may change mitochondrial membrane potential by up-regulating the protein expression of P53 and downregulating the protein expression of Bcl-2,and activate the expression of Caspase-3,c-PARP and Caspase-9,so as to activate endogenous mitochondrial pathway and promote the apoptosis of HT-29 cells.

关 键 词：伪士的宁 结肠癌 HT-29细胞 凋亡 线粒体跨膜电位 机制 P53 CASPASE-3 CASPASE-9 兔源DNA修复酶 Bcl-2 

分 类 号：R285.5[医药卫生—中药学]