干蟾皮中酯蟾毒配基和华蟾酥毒基的纯化及其体外抗结肠癌活性的研究  

Isolation and Anti-Colon Activity in Vitro of Resibufogenin and Cinobufaginin from Toad Skin

作　　者：唐晓霞[1] 贾燕丽 田港[1] 宋霄宏[1] 

机构地区：[1]浙江中医药大学药学院,杭州浙江310053

出　　处：《中华中医药学刊》2015年第9期2243-2245,共3页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省科技厅项目(2013C33179);浙江省大学生科技创新活动计划暨新苗人才计划项目(2014R410054)

摘　　要：目的：探讨干蟾皮中酯蟾毒配基和华蟾酥毒基的分离和纯化方法,并考察其体外抗结肠癌活性。方法：采用硅胶柱层析法纯化干蟾皮中的酯蟾毒配基和华蟾酥毒基,考察径高比、上样量及洗脱曲线对酯蟾毒配基和华蟾酥毒基纯化的影响;采用MTT法测定纯化产物对结肠癌细胞CT-26的生长抑制作用,考察其体外抗结肠癌活性。结果：纯化条件径高比为2∶14,上样量为10g（干蟾皮原生药材）,洗脱溶剂用量为160m L。在9.32~300μg/m L浓度范围内纯化产物对结肠癌细胞有较强的抑制作用,并且呈时间和剂量依赖关系。结论：硅胶柱层析法适合于干蟾皮中脂蟾毒配基和华蟾酥毒基的分离、纯化,且纯化产物体外抗结肠癌作用明显。Objective： To isolate resibufogenin and cinobufaginin from toad skin in order to investigate their anti-colon activity in vitro. Methods： Purify resibufogenin and cinobufagin in toad skin by silica gel column chromatography purification and inspect the ratio of diameter to height,sample volume and elution curve. Their anti-colon activity of CT-26 in vitro was evaluated by MTT colorimetric method for colon cancer cell growth inhibitory effect and we investigated its activity against colon cancer in vitro. The cytotoxicity of resibufogenin and cinobufaginin against CT-26 cell was determined using the MTT assays. Results： The purification conditions of diameter to height ratio was 2 ∶ 14,sample amount for10 g（toad skin）,elution solvent dosage 160 m L. Purification products had strong inhibition on the colon cancer cells in time and dose dependent in the rage of 9. 32 μg/m L ~ 300 μg/m L. Conclusion： The silica gel column chromatoraphy is suitable for purification of resibufogenin and cinobufaginin and its constituents have prominent anti-colon activity.

关 键 词：干蟾皮 酯蟾毒配基 华蟾酥毒基 纯化 抗结肠癌活性 

分 类 号：R285.5[医药卫生—中药学]