化学学报 ›› 2013, Vol. 71 ›› Issue (05): 722-728.DOI: 10.6023/A13010109 上一篇    下一篇

研究论文

桑白皮水提取多糖组分的分离纯化和结构特征

张晓曼, 廖文锋, 丛启飞, 董群, 丁侃   

  1. 中国科学院上海药物研究所 上海 201203
  • 投稿日期:2013-01-21 发布日期:2013-03-15
  • 通讯作者: 董群, qdong@mail.shcnc.ac.cn;丁侃,kding@mail.shcnc.ac.cn E-mail:kding@mail.shcnc.ac.cn
  • 基金资助:

    项目受国家自然科学基金(No. 30870551)及新药创制重大专项(No. 2012ZX09301001-003)资助.

Isolation and Structural Characterization of the Polysaccharides of Cortex Mori radices

Zhang Xiaoman, Liao Wenfeng, Cong Qifei, Dong Qun, Ding Kan   

  1. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203
  • Received:2013-01-21 Published:2013-03-15
  • Supported by:

    Project supported by the National Natural Science Foundation of China (No. 30870551) and the Key New Drug Creation and Manufacturing Program (No. 2012ZX09301001-003).

桑白皮为桑科植物桑Morus alba L的干燥根皮, 是临床上常用的中药. 采用沸水提取获得桑白皮中的多糖, 用乙醇沉淀、阴离子交换柱层析、凝胶过滤层析等方法进行分离和纯化, 从水提粗多糖中得到均一多糖CMA-a-1, CMA-a-5和CMA-b1-1. 应用糖组成分析、甲基化分析及IR, NMR等光谱学方法研究桑白皮多糖的结构及性质. 结果表明, CMA-a-1, CMA-a-5均为淀粉类多糖, 前者结构类似于支链淀粉, 但O-6位上取代的支链中含有1,6-连接葡萄糖, 平均链长为23; 而后者更接近于直链淀粉, 平均链长为30. CMA-b1-1为以1,2-连接的鼠李糖及1,4-连接的半乳糖醛酸为主链的RG-I型果胶类多糖, 具有多种形式的由阿拉伯糖、半乳糖、木糖等构成的支链. CMA-b1-1对SMMC7721肝癌细胞生长具有抑制作用, 对L02正常肝细胞生长则没有抑制作用. 本研究表明桑白皮水提多糖中主要是淀粉类和果胶类多糖.

关键词: 桑白皮, 桑, 多糖, 化学结构, 抗肿瘤

Cortex mori radices, the dried root cortex of Morus alba L., is a clinically widely used traditional Chinese medicine, with the effects of anti-tussive, preventing asthma, hypoglycemic, antitumor, antibacterial and antioxidative. Up to now, its polysaccharides and their contribution to biological activities remain largely unknown. In this study, the root cortex was extracted with boiling water, followed by ethanol precipitation, fractionation and purification by anion-exchange chromatography on DEAE-cellulose column and gel permeation chromatography on Sephacryl S-300 column. Three polysaccharides, CMA-a-1, CMA-a-5 and CMA-b1-1, were isolated from boiling-water extract, and characterized to be homogeneous fractions by high performance gel permeation chromatography (HPGPC). Their structural features were elucidated by sugar analyses, methylation analyses, IR, and NMR spectroscopic methods. Both CMA-a-1 and CMA-a-5 are found to be starch fractions. CMA-a-1, with an average molecular weight (MW) of 224 kDa, is an amylopectin-like polysaccharide with an average chain length (CL) of 23, containing 1,4-linked α-D-glucopyranosyl backbone, with 1, 6-linked glucose in the side chains attached to O-6 of backbone residues. CMA-a-5, with a MW of 15.5 kDa, is mainly an amylose fraction with an average chain length of 30. CMA-b1-1 was isolated from 0.2 mol·L-1 NaCl-eluted fraction upon anion-exchange chromatography, and characterized as a RG-I type pectic polysaccharide, which contains a backbone composed of alternating 1,4-linked α-D-galacturonic acid and 1,2-linked α-L-rhamnose residues, with various branches attached to O-4 of rhamnose residues, comprising of T-, 1,3-, 1,5-, 1,3,5-linked Araf, T-,1,6-, 1,3,6-linked Galp, 1, 4-linked Xylp, and 1, 4-linked Glcp. MTT method showed that CMA-b1-1 could inhibit the proliferation of SMMC7721 liver cancer cells and showed no impairment to L02 normal liver cells. This study suggested that the boiling water extract of cortex mori radices contains mainly starch and pectic fraction as the major component polysaccharides, and that the RG-I polysaccharide in vitro can inhibit the liver cancer cell proliferation, but the starch fractions cannot.

Key words: cortex mori radices, Morus alba L., polysaccharide, chemical structure, antitumor