摘要
目的:探讨植物钩吻中生物碱单体对肝癌细胞HepG2增殖的抑制作用,并探讨其相关机制。方法:MTT法观察3种生物碱单体对HepG2细胞的体外杀伤作用,选取活性最好的单体进一步行时效、量效关系的测定和细胞形态学观察;流式细胞术检测细胞周期和凋亡率的变化;Capspase-3、Capspase-8、Capspase-9活性检测用分光光度法进行。结果:3种单体都显示出对HepG2细胞的增殖抑制活性,其中钩吻素己效果最好,呈时间及剂量依赖性抑制HepG2细胞增殖;经钩吻素己处理48 h后的HepG2细胞变小变圆,正常贴壁细胞随着药物浓度的增加而减少;各浓度组钩吻素己可阻滞细胞于S期,并诱导细胞凋亡,且Capspase-3、Capspase-8、Capspase-9活性随着药物浓度的增高而增高。结论:3种钩吻生物碱单体中钩吻素己能显著抑制HepG2细胞体外增殖且细胞毒性较小,并通过影响细胞周期分布和激活Capspase-8、Capspase-9进而活化Capspase-3发挥抗肿瘤作用。
Objective：To investigate the effects of alkaloid monomers from Gelsemium elegans on proliferation of HepG2 cell in vitro and the possible mechanism.Methods：MTT assay was used to measure the inhibitory of three alkaloid monomers on HepG2 cell in vitro.The most effective fraction was chosen to test whether the effect was in time-and dose-dependent manner.The morphological changes were observed by the light microscope and the cell cycle alteration through the flow cytometric assay.The activity of Caspase-3,Caspase-8 and Caspase-9 were detected by a Caspases colorimetric assay kit.Results：The results showed that koumine,Gelsemine and Gelsenicine could significantly inhibit the proliferation of HepG2 cell and Gelsenicine,the most effective fraction,was clearly in dose-and time-dependent manners,while exhibited low cytotoxicity to the Vero cell.The cell treated with Gelsenicine for 48 h showed distinctive morphological changes.The cells treated with 200 and 400 μg/mL shrinked and fell off from the bottom.At the same time,the cells were arrested at S phase and the apoptosis increased apparently.The activity of Caspase-3,Caspase-8 and Caspase-9 was increased in a dose-dependent manner.Conclusion：Three alkaloid monomers from Gelsemium elegans,especially Gelsenicine,could inhibit proliferation of HepG2 cell obviously.The mechanism may be related to cell cycle arrest and activation of Caspase-3,Caspase 8 and Caspase-9.
出处
《中药材》
CAS
CSCD
北大核心
2012年第3期438-442,共5页
Journal of Chinese Medicinal Materials
基金
广东省自然科学基金团队项目(8351063201000003)
