天然产物研究与开发 ›› 2025, Vol. 37 ›› Issue (1): 40-51.doi: 10.16333/j.1001-6880.2025.1.005 cstr: 32307.14.1001-6880.2025.1.005

• 研究论文 • 上一篇    下一篇

醋柴胡多糖的分离纯化及协同抗肝癌活性的研究

刘志芬1,王小双2,李   苹1,吴亚运1,刘丽娟2,赵瑞芝2,赵   亚1*   

  1. 1广州中医药大学第二临床医学院 中医证候全国重点实验室;2广州中医药大学第二临床医学院 省部共建中医湿证重点实验室,广州 510006
  • 出版日期:2025-01-22 发布日期:2025-01-22
  • 基金资助:
    国家自然科学基金面上项目(82074086,82173981)

Isolation,purification and synergistic anti-liver cancer activity of polysaccharide from vinegar-baked Bupleuri Radix

LIU Zhi-fen1,WANG Xiao-shuang2,LI Ping1,WU Ya-yun1,LIU Li-juan2,ZHAO Rui-zhi2,ZHAO Ya1*   

  1. 1National Key Laboratory of Chinese Medicine Syndrome,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine;2State Key Laboratory of Dampness Syndrome of Chinese Medicine,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510006,China
  • Online:2025-01-22 Published:2025-01-22

摘要:

对醋柴胡多糖(polysaccharide from vinegar-baked Bupleuri Radix,VBRBP)进行分离纯化,初步分析其结构特征并探究其联合化疗药的协同抗肝癌活性。通过水提醇沉、膜分离及纤维素层析从醋柴胡中分离纯化得到醋柴胡均一多糖,采用高效凝胶渗透色谱法、PMP柱前衍生化法、傅里叶变换红外光谱、气质联用和核磁共振波谱等方法对醋柴胡均一多糖进行结构鉴定,并测定其联合不同化疗药物对HepG2和Huh7两种肝癌细胞株的增殖抑制作用。结果表明,经过分离纯化得到的醋柴胡均一多糖5-1(polysaccharide 5-1 from vinegar-baked Bupleuri Radix ,VBRBP5-1)是一种中性多糖,相对分子量为30.78 kDa,由甘露糖、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖组成,摩尔比为1.96∶2.38∶58.48∶22.85∶14.32;主链是由1-α-D-葡萄糖、1,6-α-D-葡萄糖、1,4,6-α-D-葡萄糖、1,4-α-D-葡萄糖、1,3,6-β-D-半乳糖残基构成,支链主要含有1-β-D-半乳糖、1,4-β-D-半乳糖和1-α-L-阿拉伯糖、1,3,5-α-L-阿拉伯糖、1,5-α-L-阿拉伯糖残基片段。VBRBP5-1能增强10-羟基喜树碱、甲氨蝶呤和顺铂对肝癌细胞的抑制率,显示出协同增效作用,其中对顺铂的协同增效作用在两种肝癌细胞中均优于其他抗肿瘤药物。本研究解析了VBRBP5-1的结构并探索了其协同抗肝癌活性,为醋柴胡多糖的构效关系阐释及后续深入开发和利用提供参考。

关键词: 醋柴胡, 均一多糖, 结构表征, 协同抗肝癌

Abstract:

This study aims to isolate and purify polysaccharide from vinegar-baked Bupleuri Radix(VBRBP),and to preliminarily analyze its structural characteristics,and explore its synergistic anti-liver cancer activity in combination with chemotherapeutics.One homogeneous polysaccharide from vinegar-baked Bupleuri Radix was isolated and purified by water extraction and alcohol precipitation,membrane separation and cellulose chromatography.High-performance gel permeation chromatography,1-phenyl-3-methyl-5-pyrazolon pre-column derivatization,Fourier transform infrared spectroscopy,gas chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy were used to identify the structure of VBRBP,and the inhibitory effect of its combination with different chemotherapeutics on the proliferation of HepG2 and Huh7 liver cancer cell lines was measured.The results showed that the homogeneous polysaccharide 5-1 from vinegar-baked Bupleuri Radix (VBRBP5-1) obtained through separation and purification was a neutral polysaccharide with a relative molecular weight of 30.78 kDa,composed of mannose,galacturonic acid,glucose,galactose and arabinose,with a molar ratio of 1.96∶2.38∶58.48∶22.85∶14.32.The backbone of VBRBP5-1 was composed of 1-α-D-glucose,1,6-α-D-glucose,1,4,6-α-D-glucose,1,4-α-D-glucose and 1,3,6-β-D-galactose residues,and the branched chain mainly contained 1-β-D-galactose,1,4-β-D-galactose and 1-α-L-arabinose,1,3,5-α-L-arabinose,1,5-α-L-arabinose residues. Combined administration of VBRBP5-1 could enhance the inhibition rates of 10-hydroxycamptothecine,methotrexate and cisplatin on hepatocellular carcinoma cells,showing a synergistic effect,in which the synergistic effect on cisplatin was superior to the others in these two hepatocellular carcinoma cells.In this study,the structure of VBRBP5-1 was analyzed and its synergistic anti-liver cancer activity was explored,which provided a reference for the interpretation of the structure-activity relationship and subsequent further development and utilization of polysaccharides from vinegar-baked Bupleuri Radix.

Key words: vinegar-baked Bupleuri Radix, homogeneous polysaccharide, structural characterization, synergistic anti-liver cancer

中图分类号:  R284