仿刺参(Apostichopus japonicus)中皂苷类成分的分离鉴定及细胞毒活性研究  

Study on isolation and tumor-cytotoxic activity of triterpene glycosides in Apostichopus japonicus

作　　者：徐蕾[1] 宋清平 杨爽[1] 于明明[1] 王远红[1] 姜廷福[1] 吕志华[1,2] XU Lei;SONG Qing-ping;YANG Shuang;YU Ming-ming;WANG Yuan-hong;JIANG Ting-fu;LV Zhi-hua(Key Laboratory of Marine Drugs,Ministry of Education,Shandong Provincial Key Laboratory of Glycoscience and Glycoengineering,School of Medicine and Pharmacy,Ocean University of China, Qingdao 266003,China;Laboratory of Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology,Qingdao 266237,China)

机构地区：[1]中国海洋大学海洋药物教育部重点实验室,山东省糖科学与糖工程重点实验室,医药学院,山东青岛266003 [2]青岛海洋科学与技术国家实验室海洋药物与生物制品功能实验室,山东青岛266237

出　　处：《中国海洋药物》2018年第6期38-46,共9页Chinese Journal of Marine Drugs

基　　金：国家科学基金委员会-山东省人民政府联合基金海洋药物与生物制品项目(U1606403);国家海洋局海洋公益性行业科研专项项目(201405038);国家自然科学基金青年科学基金项目(31700704);中国博士后科学基金第58批面上资助项目(2015M582145);山东省自然科学基金博士基金项目(ZR2017BC092);青岛海洋科学与技术国家实验室鳌山科技创新计划项目(2015ASKJ02)资助

摘　　要：目的对仿刺参(Apostichopus japonicus)体内皂苷类成分进行分离及结构鉴定,研究纯化皂苷的抗肿瘤细胞毒活性。方法以鲜仿刺参为原材料,选用60%的乙醇溶液从海参中提取出海参皂苷,然后通过有机溶剂萃取法、大孔树脂法、硅胶柱层析、凝胶柱层析、半制备HPLC分离纯化海参皂苷,采用LTQ Orbitrap XL质谱、Jeol JNM-ECP 600核磁技术对得到的海参皂苷进行结构鉴定,以人肺癌细胞(A549)、人结肠癌细胞(HCT116)、人肝癌细胞(BEL-7402)、人肺腺癌细胞(NCI-H1975)和正常肝细胞(L-02)为对象对纯化皂苷抗肿瘤细胞毒活性进行研究。结果从鲜仿刺参同时得到3种三萜皂苷类化合物,分别为Holotoxin D、Holotoxin B1和Holotoxin A1。3种皂苷的细胞毒活性具有一致性,对肿瘤细胞具有良好的抑制率,抑制率均随皂苷单体浓度增大而增大,且到达一定浓度后趋于稳定,均呈现弱效抑制作用。结论从鲜仿刺参中同时纯化得到Holotoxin D,Holotoxin B1和Holotoxin A1,比例为1∶7.97∶14.94。3种皂苷单体具有良好的抗肿瘤活性,Holotoxin D抗肿瘤活性为首次报道,本研究为以后的肿瘤药物的筛选研究奠定了良好的基础。Objective The triterpene glycosides from the Apostichopus japonicus were isolated and identified,and tumor-cytotoxic activities of the compounds were further determined.Methods The triterpene glycosides were extracted from fresh Apostichopus japonicus with 60% aq.ethanol.The triterpene glycosides were separated and purified by organic solvents,macroporous resin,silica gel column chromatography,gel column chromatography and semi preparative HPLC.The structures were determined by ESI-MS and NMR,the tumor-cytotoxic activities were studied with A549 cells,HCT116 cells and BEL-7402cells.by the SRB assay.Results Three kinds of three triterpene glycosides were purified and identified,which were Holotoxin D,Holotoxin B1 and Holotoxin A1,respectively.The cytotoxicity of three saponins was consistent,and had a good inhibitory effect on A549,HCT116 and BEL-7402.The inhibition rates increased with the increase of saponin monomer concentration,and tended to be stable after reaching a certain concentration,showing weak inhibition.Conclusion Holotoxin D,Holotoxin B1 and Holotoxin A1 were purified from the Apostichopus japonicus for the first time synchronously,and the ratio was 1:7.97:14.94.The three kinds of saponins had good antitumor activity,and the anti tumor activity of Holotoxin D was reported for the first time.This study laid a good foundation for screening anti-tumor drugs in the future.

关 键 词：仿刺参 海参皂苷 分离鉴定 细胞毒 

分 类 号：R917[医药卫生—药物分析学]