白头翁皂苷D体外抗肝癌作用及其机制研究  

Antihepatocarcinoma effect in vitro of pulchinenoside D and mechanisms

作　　者：岳文华[1,2] 徐坤 冯育林[1] 张天[2] 许琼明[2] 杨世林[1,2] 刘艳丽[2] 

机构地区：[1]江西中医药大学药学院,江西南昌330000 [2]苏州大学药学院,江苏苏州215123 [3]铜仁市人民医院药学部,贵州铜仁554300

出　　处：《中草药》2014年第22期3295-3301,共7页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81273402);重大新药创制国家科技重大专项资助(2011ZX11102-001-19);江苏省自然科学基金资助(BK2012620);2013年苏州大学大学生创新创业训练计划(2013xj060)

摘　　要：目的探讨白头翁皂苷D体外抗肝癌作用及其相关机制。方法采用MTT法考察白头翁皂苷D对人肝癌细胞BEL-7402增殖的影响,同时观察BEL-7402细胞形态的变化,采用集落形成实验考察白头翁皂苷D对BEL-7402细胞集落形成的影响;Hoechest 33342染色观察BEL-7402细胞凋亡的形态变化;流式细胞术检测BEL-7402细胞凋亡率和线粒体膜电位的变化;采用Western blotting法检测BEL-7402细胞Bcl-2和Caspase-3蛋白的表达。结果 MTT结果显示白头翁皂苷D对BEL-7402细胞增殖有抑制作用;集落形成实验结果显示白头翁皂苷D能明显抑制BEL-7402细胞集落的形成,白头翁皂苷D(25.00μg/m L)能显著诱导BEL-7402细胞凋亡;白头翁皂苷D(18.75、25.00μg/m L)能显著降低BEL-7402细胞线粒体膜电位;白头翁皂苷D(12.50、18.75、25.00μg/m L)可显著上调BEL-7402细胞中Caspase-3的表达;白头翁皂苷D(25.00μg/m L)可显著下调Bcl-2的表达。结论白头翁皂苷D有良好的体外抗肝癌作用,其机制与调节线粒体途径凋亡相关蛋白Bcl-2、Caspase-3的表达有关。Objective To research the antihepatocarcinoma effect in vitro ofpulchinenoside D and mechanisms. Methods MTT assay was used to evaluate the proliferative inhibition of pulchinenoside D on hepatoma carcinoma cells （BEL-7402）, and the morphology of BEL-7402 cells was also observed; Colony formation method was used to examine colony-forming role by pulchinenoside D on BEL-7402 cells; The morphological changes of drug-induced apoptosis of BEL-7402 were observed by Hoechst 33342 staining; Flow cytometry assay was used to analyze the apoptosis rate and mitochondrial membrane potential changes of BEL-7402 cells; Westem blotting was employed to determine the expression levels of Bcl-2/Caspase-3 protein in BEL-7402 cells. Results MTT assay showed that pulchinenoside D could inhibit the proliferation of BEL-7402 cells, and colony formation method showed that pulehinenoside D could significantly inhibit the colony formation of BEL-7402 cells; Pulchinenoside D （25.00 μg/m） could significantly induce the apoptosis of BEL-7402; Pulchinenoside D （18.75 and 25.00 μg/m） could significantly reduce the mitochondrial membrane potential of BEL-7402 cells; Pulchinenoside D （12.50, 18.75, and 25.00μg/m） could significantly increase the expression of Caspase-3; Pulchinenoside D （25.00 μg/m） could significantly down-regulate the expression of Bcl-2. Conclusion Pulchinenoside D has antiturnor effects in vitro; The mechanism is involved in regulation of the mitochondrial pathway apoptosis-related protein expression of Bcl-2 and caspase-3.

关 键 词：抗肿瘤 肝癌 细胞凋亡 线粒体途径 

分 类 号：R286.91[医药卫生—中药学]