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牛耳枫生物碱2-hydroxyyunnandaphnine D体外抗肿瘤活性及作用机制 预览 被引量:3

Anti-tumor activity and the mechanism of 2-hydroxyyunnandaphnine D from Daphniphyllum calycinum in vitro
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摘要 为了研究牛耳枫生物碱2-hydroxyyunnandaphnine D体外抗肿瘤活性及作用机制,用MTT法检测6.25~100mg/L2-hydroxyyunnandaphnine D分别作用于人肝癌细胞(HepG-2)、人乳腺癌细胞(MCF-7)及人宫颈癌细胞(HeLa)后不同时间点的细胞存活并计算细胞增殖抑制率和IC50;FDA/PI双染色荧光显微镜观察细胞死亡情况;分光光度法检测Caspase-3酶活性变化;MTT法检测凋亡抑制剂Z-VAD-FMK对2-hydroxyyunnandaphnine D抑瘤活性的影响.结果显示:2-hydroxyyunnandaphnine D对3种瘤株均有明显的抑制作用,且具有明显的时效量效关系,其48h的IC50值分别为(1.30±0.09),(7.32±0.10)和(8.41±0.11)mg/L.FDA/PI双染色荧光显微镜观察显示2-hydroxyyunnandaphnine D组中死亡细胞数较正常对照组细胞死亡数量增加,且随着剂量的加大细胞死亡数量增加.MTT法显示凋亡抑制剂Z—VA胁FMK不能阻断2个浓度的2-hydroxyyunnandaphnine D对HepG-2细胞的增殖抑制作用.Caspase-3酶活性测定显示10,30mg/L2-hydroxyyunnandaphnine D作用后肿瘤细胞的Caspase-3酶活力单位与正常对照组比较无统计学意义.结果表明:牛耳枫生物碱2-hydroxyyunnandaphnine D在体外具有明显的抑制肿瘤细胞增殖的作用,其作用机制并非通过激活Caspase-3途径. To study the anti-tumor activity and the mechanism of 2-hydroxyyunnandaphnine D from Daphniphyllum calycinum. The proliferation inhibition rate and IC50 of HepG-2, MCF-7 and HeLa cells were measured by MTT assay in vitro treated by 2-hydroxyyunnandaphnine D(6.25 - 100 mg/L)after 24 , 48 and 72 h. The cell death was observed by fluorescence microscopy . The inhibition pathway of 2-hydroxyunnandaphnine D was analyzed by apoptosis inhibitor Z-VAD-FMK by MTT. The activity of Caspase-3 in tumor cells was detected by spectrophotography. The MTT assay results revealed that 2- hydroxyyunnandaphnine D could inhibit the proliferation of the three kinds of tumor cells obviously and the inhibitory rates significantly increased in a dose-dependent and time-dependent manner. The IC50 were (1.30±0.09), (7.32±0.10)and(8.41±0.11)mg/L after 48 h treated with 2-hydroxyyunnandaphnine D respectively. There were more death cells in 2-hydroxyyunnandaphnine D groups than that of normal group. The killing effect on HepG-2 cells of 2-hydroxyyunnandaphnine D was not impacted by Z-VADFMK. The Caspase-3 enzyme activity of HepG-2 cells was no obvious change compared with the control group, which indicated that Caspase-3 had not been activated in the course of cell death induced by 2- hydroxyyunnandaphnine D. The results suggested that 2-hydroxyyunnandaphnine D could inhibit the proliferation of tumor cells and the mechanism were not by activating Caspase-3.
作者 王蓓 戎瑞雪 郑聪毅 曹志然 罗都强 WANG Bei , RONG Ruixue , ZHENG Congyi , CAO Zhiran , LUO Duqiang (1. College of Basic Medical Science, Hebei University, Baoding 071000, China; 2. College of Life Sciences, Hebei University, Baoding 071002, China)
出处 《河北大学学报:自然科学版》 CAS 北大核心 2013年第4期401-407,共7页 Journal of Hebei University (Natural Science Edition)
基金 基金项目:国家自然科学基金资助项目(30671385)
关键词 牛耳枫 2-hydroxyyunnandaphnine D MTT Z-VAD—FMK CASPASE-3 Daphniphyllum calycinum 2-hydroxyyunnandaphnine D MTT Z-VAD-FMK Caspase-3
作者简介 王蓓(1978-),女,河北献县人,河北大学讲师,硕士,主要从事肿瘤免疫及抗感染免疫研究.E-mail:wp780203@163.com 通信作者:曹志然(1963-),女,河北景县人,河北大学教授,主要从事中药药理抗肿瘤研究.E-mail:caozhiran@163.com 通信作者:罗都强(1968-),男,陕西扶风人,河北大学教授,博士,主要从事天然药物性成分分析.E-mail:Duqiangluo@163.com
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