王不留行中抗人微血管内皮细胞增殖的有效组分的分离和纯化  

Isolation,Purification and Identification of Chemical Compound from Semen vaccariae to Inhibit Endothelial Cells

作　　者：夏明星 冯磊 张莲芬 李英 储敏 金坚 

机构地区：[1]江南大学医药学院细胞与分子药理实验室,无锡214122

出　　处：《生物技术通报》2009年第2期93-97,共5页Biotechnology Bulletin

基　　金：国家863计划项目(2006AA02Z153);上海市科委生物医药重大科技攻关项目(06DZ19020)

摘　　要：从中药王不留行中筛选具有抑制内皮细胞增殖的有效单一组分。大孔树脂吸附法进行总皂甙的分离纯化,分别收集不同浓度乙醇溶液洗脱液用SRB法进行细胞活性检测,选择有效部分用AKTA Resource柱分离,按紫外吸收峰收集洗脱液并进行活性验证,通过蒸发光散射检测组分复杂程度,选择活性高、丰度大、成份简单的峰进行C18柱分离。结果表明,王不留行提取液经大孔树脂分离和细胞活性检测发现50%～90%乙醇浓度洗脱液具有细胞活性,经AKTA Resource柱分离得7种组份群(0～6),细胞活性测得3～6号组分有活性,选择4号组分过C18柱得5组分(A～E),B组分为有紫外吸收的主峰,但无细胞活性,其活性分布于D和E组分中,其IC50值为3.75μg/ml,D组分经蒸发光散射检测为单峰。经上述方法分离纯化获得抑制人微血管内皮细胞增殖的单一组分群,其单一化合物具有潜在的利用价值。The study aimed at screening a unity chemical compound from semen vaccariae to inhibit proliferation and growth of endothelial cells. Isolation and purification of total saponin in macroporous resin,and gradient elution by the alcohol with diversity of concentration were carried out. The utility region was isolated by AKTA Resource column, where the elution was collected based on ultraviolet absorption. Meanwhile,detecting the elution by ELSD and choosing the fraction with high activity,abundance and reconstituent simply to further segregation were accomplished. Finally,the unity chemical compound were obtained by C18 column. Results showed that,semen vaccariae extracts were isolated by macroporous resin,with 50%-90% alcohol concentration ehition that have the activity to inhibit the endothelial cells. Seven groups （0-6） were obtained,and the 3-6 samples were proved to have the capability to inhibit the target cells. Five groups （A-E） were further separated from sample 4 where the B groups were the main peak with obvious ultraviolet absorption,but no inhibition on endothelial cells,D and E groups that had the activity in ECs. Thus,the homogeneous chemical which can inhibit proliferation and growth of endothelial cells compound had been discovered.

关 键 词：王不留行 内皮细胞 分离纯化 

分 类 号：R284[医药卫生—中药学]