基于中药整体性的红豆杉抗乳腺癌活性成分筛选方法研究  

Study on methods for screening anti-tumor active components of Taxus chinensis based on integrity of traditional Chinese medicine

作　　者：薛雪 朱龙平[2] 杨得坡[2] 赵志敏[2] XUE Xue;ZHU Long-ping;YANG De-po;ZHAO Zhi-min(Pharmacy Department of School of Pharmaceutical Science,Guangdong Food and Drug Vocational College,Guangzhou 510520,China;Laboratory of Raw Medicine and Natural Medicine,School of Pharmaceutical Science,Sun Yat-sen University,Guangzhou,510006,China)

机构地区：[1]广东食品药品职业学院药学院药学教研室,广东广州510520 [2]中山大学药学院生药与天然药化实验室,广东广州510006

出　　处：《中草药》2020年第24期6278-6285,共8页Chinese Traditional and Herbal Drugs

基　　金：广东省中医药局科研项目(20201245);广东省中医药局科研项目(20181188);广东省中医药局科研项目(20211284);广东省医学科研基金(B2018258);广东省重点平台和重大科研项目(2017GkQNCX036)。

摘　　要：目的基于中药整体性建立2种红豆杉抗乳腺癌活性成分筛选方法:中空纤维细胞捕获结合高效液相色谱法(HFCF-HPLC)、细胞捕获-分散液相微萃取结合高效液相色谱法(CF-DLLME-HPLC)。方法 HFCF-HPLC是在模拟人体内环境状态下,将人乳腺癌MCF-7细胞种植到聚丙烯中空纤维管腔内部,再将该中空纤维放入红豆杉提取液中,筛选和捕获可以与细胞结合的抗乳腺癌活性成分群;CF-DLLME-HPLC是将MCF-7细胞与红豆杉提取液在细胞正常代谢的基础上共孵育,取不同孵育时间上清液进行分散液相微萃取处理,然后进行不同时间段下色谱分析比对,根据色谱峰变化发现潜在抗乳腺癌活性成分群。结果 HFCF-HPLC筛选出的活性成分是脱乙酰巴卡亭、巴卡亭Ⅲ、紫杉碱M、去乙酰紫杉醇、三尖杉宁碱、紫杉醇、云南紫杉烷;CF-DLLME-HPLC筛选出的活性成分是脱乙酰巴卡亭、巴卡亭Ⅲ、紫杉碱M、去乙酰紫杉醇、金松双黄酮。结论 2种方法均可以筛选出有效的抗肿瘤活性成分群,为活性成分的筛选、中药质量的综合评价提供一种方便、普适和高效的新方法。Objective Two screening methods for drug active constituents were established based on the integrity of traditional Chinese medicine to screen the anti-breast cancer active components, which were hollow fiber cell fishing-high performance liquid chromatography(HFCF-HPLC) and cell fishing-dispersive liquid-liquid microextraction-high performance liquid chromatography(CF-DLLME-HPLC). Methods According to HFCF-HPLC, MCF-7 cells were planted into the cavity of polypropylene hollow fiber, and then the hollow fiber was put into the extract of Taxus chinensis to screen and capture the bioactive component groups combined with cells in a simulated human body environment. According to CF-DLLME-HPLC, on the basis of normal cell metabolism, MCF-7 cells were co-incubated with T. chinensis extract, supernatant of different incubation time was taken for liquid phase microextraction, and then chromatographic analysis was performed to find potential active component groups according to chromatographic peak changes. Results Anti-breast cancer active components of T. chinensis were obtained by these two methods. 10-Deacetylbaccatin III, baccatin Ⅲ, taxinine M, 10-deacetyltaxol, cephalomannine, taxol, taxuyunnanine C were obtained by HFCF-HPLC, and 10-deacetyltaxol, baccatin Ⅲ, taxinine M, 10-deacetyltaxol, sciadopitysin were obtained by CF-DLLME-HPLC. Conclusion The two methods can screen the effective antitumor active ingredients, and provide a convenient, universal and efficient new method for the screening of active ingredients and the comprehensive evaluation of the quality of traditional Chinese medicine.

关 键 词：红豆杉 抗乳腺癌 筛选方法 活性成分群 中药整体性 中空纤维细胞捕获结合高效液相色谱法 细胞捕获-分散液相微萃取结合高效液相色谱法 脱乙酰巴卡亭 巴卡亭Ⅲ 紫杉碱M 去乙酰紫杉醇 三尖杉宁碱 紫杉醇 云南紫杉烷 金松双黄酮 

分 类 号：R285.5[医药卫生—中药学]