【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。更多还原

【Abstract】 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.更多还原