【作者】 刘艳丽； 季秀美； 徐坤； 舒展； 何文君； 潘舒； 许琼明； 李笑然；

【Author】 Liu Yanli,Ji Xiumei,Xu Kun,Shu Zhan,He Wenjun,Panshu,Xu Qiongming,Li Xiaoran College of Pharmacy,Soochow University,Suzhou 215123,China

【机构】 苏州大学药学院；

【摘要】 目的:探讨白头翁总皂苷及单体的抗肿瘤作用。方法:体外MTT法考察白头翁皂苷对SMMC-7721细胞增殖的抑制作用;采用细胞集落形成实验考察其克隆形成率;流式细胞仪和投射电镜分析细胞凋亡;小鼠H₂₂和S₁₈₀模型考察白头翁皂苷的抗肿瘤作用。Westblotting考察凋亡相关蛋白的表达。结果:白头翁总皂苷通过诱导细胞凋亡而抑制肿瘤的生长;H₂₂和S₁₈₀移植瘤小鼠给药10d后,给药组瘤重明显降低,抑瘤率分别达到43.7%（200 mg/kg）及49.8%（200 mg/kg）;采用天然药物化学分离方法,从白头翁总皂苷中分离、鉴定和富集36个单体皂苷,其中30种齐墩果酸型皂苷,6种白桦脂酸型皂苷。采用MTT法,发现其中3种皂苷类成分（化合物5,6,10）体外抗肿瘤作用最好;MTT和集落实验提示化合物5,6和10对人肝癌细胞SMMC-7721细胞和BEL-7402细胞的增殖有明显的抑制作用;化合物5和6（6 mg/kg）对小鼠H₂₂移植瘤有明显的抑制作用;化合物6（25μg/ml）和5（6.26μg/ml）诱导SMMC-7721细胞凋亡。化合物10降低bcl-2的表达,升高P53的表达。结论:白头翁总皂苷有抗肿瘤作用,其作用与诱导细胞凋亡有关,化合物5,6,10可能是其主要的活性成分。更多还原

【Abstract】 Aim:The present study was undertaken to investigate if Pulsatilla chinensis saponins（PRS） possesses antitumor effect,Structure-Activity Relationship of saponins and antitumor mechanism tumor cells in vitro and vivo. Materials and Methods:The in vitro effect of PRS against tumor cells was mesure by MTT assay. The effects of PRS on cell cycle and cell apoptosis was analyzed by flow cytometry.The in vivo antitumor activity was observed in H₂₂ tumor bearing mice.Results:PRS inhibited the proliferation of human tumor cells in vitro by apoptosis.10 days after administration of PRS（50,100,200 mg/kg）,the weight of tumor mass was markly decreased in mice.The antitumor effect of PRS in vivo was associated with a significant increase in the H₂₂ cells apoptosis rate;36 saponins isolated from PRS,including 14 Oleanolic acid glycosides,16 hederagenin glycosides,2 23-hydroxy betulinic acid glycosides and 4 betulinic acid glycosides.Compound 5,6 and 10 had the most antitumor activity against 7721 cancer cell lines.Figure 6 showed that cells were stained with Hoechst 33342;5 and 6 inhibited cell SMMC-7721 cells proliferation in a dose-dependent manner,and induced apoptosis in cells.6 or 5 induced apoptosis in 7721 cells.10 days after administration of 6（6 mg/kg）（5:6mg/kg）（50,100,200 mg/kg）,the weight of tumor mass was markly decreased in mice transplanted with H₂₂ cells.The inhibited rate of 6 was 37.8% （6:40.4%）.Inaddition,10 inhibited 7721 cells proliferation;10 induce G₂ Arrest and inhibited Bcl2 expression,increased Cyclin B and p53 expression.Conclusion:These results suggested that PRS exerted potential anticancer activity through apoptosis.Compund 5,6 and 10 might be the bioactive compounds of PRS.更多还原