基于内切-1,4-β-甘露聚糖酶水解的黄芪糖指纹图谱的建立及不同种质资源黄芪鉴别Establishment of fingerprint of Astragali Radix polysaccharides based on endo-1,4-β-mannanase hydrolysis and identification of Astragali Radix of different germplasm resources
王迪;李科;王桂臻;李震宇;秦雪梅;杜冠华;张翔;
WANG Di;LI Ke;WANG Gui-zhen;LI Zhen-yu;QIN Xue-mei;DU Guan-hua;ZHANG Xiang;Modern Research Center for Traditional Chinese Medicine,Shanxi University;College of Chemistry and Chemical Engineering,Shanxi University;Key Laboratory of chemical Biology and Molecular Engineering Ministry,Shanxi University;Institute of Materia Medica,Chinese Academy of Medical Sciences & Peking Union Medical College;University of Louisville;
摘要(Abstract):
以内切-1,4-β-甘露聚糖酶的最佳酶解条件对不同种质资源蒙古黄芪(移栽芪和野生芪)和膜荚黄芪(移栽芪和野生芪)多糖进行酶解,通过荧光辅助糖电泳获得糖指纹图谱,利用主成分分析对数据进行分析,获得区分不同种质资源黄芪的差异性糖组分。结果显示,内切-1,4-β-甘露聚糖酶酶解产物中三糖、四糖和五糖可以作为区分蒙古黄芪(移栽芪和野生芪)的差异性糖片段,五糖和六糖可以作为区分膜荚黄芪(移栽芪和野生芪)的差异性糖片段,三糖和四糖可以作为区分移栽蒙古黄芪和移栽膜荚黄芪的差异性糖片段。研究表明,内切-1,4-β-甘露聚糖酶降解的多糖产物能很好地鉴别黄芪种属(蒙古黄芪与膜荚黄芪),生长方式(移栽芪和野生芪)。该研究为黄芪药材的品质评价及活性寡糖的筛选奠定了基础。
The polysaccharides of different germplasm resources of Astragalus membranaceus var. mongholicus(cultured Astragalus Radix(RA) and natural RA) and A. membranaceus(MJ)(cultured RA and natural RA) were studied by using the optimal enzymatic conditions of endo-1,4-β-mannanase. Saccharide fingerprints were obtained for the identification and evaluation of the germplasm resources of RA by Fluorophore-assisted Carbohydrate Electrophoresis(FACE). The data were analyzed by principal component analysis to obtain the difference between RA of different germplasm resources. The results showed that trisaccharide,tetrasaccharide and pentasaccharide of endo-1,4-β-mannanase hydrolyzate could be used as the differential fragments to distinguish MG(cultured RA and natural RA); the pentasaccharide and hexasaccharide can be used as differentially expressed carbohydrate fragments that distinguish MJ(cultured RA and natural RA); the trisaccharide and tetrasaccharide can be used as the differential fragments to distinguish the cultured MG and cultured MJ. Studies have shown that polysaccharide products degraded by endo-1,4-β-mannanase can well distinguish RA species(MG and MJ),growth mode(cultured RA and natural RA). This study laid the foundation for the quality evaluation of Astragalus medicinal herbs and screening of active oligosaccharides.
关键词(KeyWords):
内切-1,4-β-甘露聚糖酶;黄芪;多糖;指纹图谱;主成分分析;种质资源
endo-1,4-β-mannanase;Astragali Radix;polysaccharide;fingerprint;principal component analysis;germplasm resources
基金项目(Foundation): 国家自然科学基金项目(31200278);; 教育部博士点新教师类基金项目(20131401120006);; 山西省优秀人才科技创新项目(201705D211020);; 山西省中药产业重点科技攻关项目(201603D311101);; 国家中药标准化项目(ZYBZH-Y-JIN-34);; 山西省科技攻关项目(2014ZD0401)
作者(Author):
王迪;李科;王桂臻;李震宇;秦雪梅;杜冠华;张翔;
WANG Di;LI Ke;WANG Gui-zhen;LI Zhen-yu;QIN Xue-mei;DU Guan-hua;ZHANG Xiang;Modern Research Center for Traditional Chinese Medicine,Shanxi University;College of Chemistry and Chemical Engineering,Shanxi University;Key Laboratory of chemical Biology and Molecular Engineering Ministry,Shanxi University;Institute of Materia Medica,Chinese Academy of Medical Sciences & Peking Union Medical College;University of Louisville;
Email:
DOI: 10.19540/j.cnki.cjcmm.20180315.003
参考文献(References):
- [1]中国药典.一部[S].2015:302.
- [2]秦雪梅,李爱平,李科,等.山西黄芪产业发展思考[J].中国中药杂志,2016,41(24):4670.
- [3]吴铭,周桃英,陈年友,等.黄芪多糖抗疲劳作用研究[J].湖北农业科学,2014,53(1):175.
- [4]许灵波.黄芪多糖在动物生产中应用的研究进展[J].中国畜牧杂志,2015,51(3):84.
- [5]Kiyohara H,Uchida T,Takakiwa A M.Different contributions of side-chainsinβ-D-(1→3,6)-galactans on intestinal Peyer's patch immunomodulation by polysaecharides from Astragalus mongholics Bunge[J].Phytochem,2009,71(2/3):280.
- [6]Cho W C,Leng K N.In vitro and in vivo immune modulating and immune restorative effects of Astragalus membranaceus[J].J Ethonpharmaeol,2007,113:132.
- [7]Li R,Chen W C,Wang W P,et al.Extraction,characterization of Astragalus polysaceharides and its immune modulating activities in rats with gastric cancer[J].Carbohyd Polym,2009,78:738.
- [8]丁喜炎,季琳,程雪,等.不同硫熏替代技术的黄芪药材对小鼠免疫功能影响研究[J].中国中药杂志,2016,41(15):2819.
- [9]Yuan Y,Sun M,Li K S.Astragalus mongholicus polysaecharide inhibits lipopolysaceharide-induced production of TNF-αand interleukin-8[J].World J Gastroentero,2009,15(29):3676.
- [10]Zhang W J,Hufnagl P,Binder B R,et al.Antiinflammatory activity of astragalosideⅣis mediated by inhibition of NF-kappa B activation and adhesion molecule expression[J].Thromb Haemostasis,2003,90(5):904.
- [11]周芹.黄芪甲苷抗凋亡作用机制的研究进展[J].中国现代应用药学,2017,34(5):783.
- [12]周妮娜,张利英,刘永琦,等.黄芪多糖对电离辐射诱发间充质干细胞基因DNA损伤的保护作用[J].中国现代应用药学,2016,33(2):139.
- [13]邓来明,肖正华,陈定宇.黄芪多糖对糖尿病足溃疡成纤维细胞AGEs及RAGEmRNA表达的影响[J].今日药学,2014,24(5):313.
- [14]余坤子,刘靖,洪浩,等.黄芪种植产地与生态环境及饮片规格的调查研究[J].中国中药杂志,2010,35(9):1112.
- [15]谢小龙,王溪森,赵利,等.黄芪种质资源研究进展[J].安徽农业科学,2005,33(1):121.
- [16]秦雪梅,李震宇,孙海峰,等.我国黄芪药材资源现状与分析[J].中国中药杂志,2013,(19):3234.
- [17]高凤兰,孙雪筠.野生与栽培黄芪主要成分的分析测定[J].黑龙江中医药,1991,3:47.
- [18]中国药典.一部[S].2010.
- [19]姜勇,金芳,鲍忠,等.不同来源黄芪药材中黄芪甲苷的定量分析[J].中国中药杂志,2006,31(11):930.
- [20]石子仪,鲍忠,姜勇,等.不同来源黄芪药材中毛蕊异黄酮葡萄糖苷和芒柄花素的定量分析[J].中国中药杂志,2007,32(9):779.
- [21]胡芳弟,封士兰,赵健雄,等.HPLC法测定黄芪中黄酮类成分和黄芪甲苷的含量[J].分析测试技术与仪器,2003,9(3):173.
- [22]寨鸿瑞,王晓文.黄芪多糖的主要作用及在临床防治上的应用[J].河南畜牧兽医:综合版,2016,(5):37.
- [23]陈辉,肖定福.黄芪多糖的生物学功能及其在畜禽生产中的应用[J].饲料研究,2017,
- [24]Guan J,Yang F Q,Li S P.Evaluation of carbohydrates in natural and cultured cordyceps by pressurized liquid extraction and gas chromatography coupled with mass spectrometry[J].Molecules,2010,15(6):4227.
- [25]黄冬兰,徐永群,陈小康,等.红外三级鉴定法快速鉴别栽培与半野生黄芪[J].光谱实验室,2010,27(3):833.
- [26]Xu J,Guan J,Chen X J,et al.Comparison of polysaccharides from different Dendrobium using saccharide mapping[J].J Pharmaceut Biomed,2011,55:977.
- [27]Guan J,Zhao J,Feng K,et al.Comparison and characterization of polysaccharides from natural and cultured Cordyceps using saccharide mapping[J].Anal Bioanal Chem,2011,399(10):3465.
- [28]庄海峰,沈建平,庄晓芬,等.黄芪多糖对粒细胞减少期侵袭性肺曲霉病SD大鼠血清半乳甘露聚糖的影响[J].浙江医学,2014,36(23):1910.
- [29]蔡孟深,李中军.糖化学[M].北京:化学工业出版社,2006.
- [30]Morcuende R,Krappe A,Hhrry V,et al.Sucrose feeding leads to increased rates of nitrate assimilation,increased rates ofα-oxoglutarate synthesis,and increased synthesis of a wide spectrum of amino acids in tobacco leaves[J].Plants,1998,206:394.
- [31]Koch K E,Ying Z,Wu Y,et al.Multiple paths of sugar-sensing and a sugar/oxygen overlap for genes of sucrose and ethanol metabolism[J].J Exp Bot,2000,51(343):417.
- [32]苏文华,张光飞,李秀华,等.植物药材次生代谢产物的积累与环境的关系[J].中草药,2005,36(9):1415.
- [33]李钦,胡继宏,高博,等.黄芪多糖在免疫调节方面的最新研究进展[J].中国实验方剂学杂志,2017,23(2):199.
- 内切-1,4-β-甘露聚糖酶
- 黄芪
- 多糖
- 指纹图谱
- 主成分分析
- 种质资源
endo-1,4-β-mannanase - Astragali Radix
- polysaccharide
- fingerprint
- principal component analysis
- germplasm resources
- 王迪
- 李科
- 王桂臻
- 李震宇
- 秦雪梅
- 杜冠华
- 张翔
WANG Di- LI Ke
- WANG Gui-zhen
- LI Zhen-yu
- QIN Xue-mei
- DU Guan-hua
- ZHANG Xiang
- Modern Research Center for Traditional Chinese Medicine
- Shanxi University
- College of Chemistry and Chemical Engineering
- Shanxi University
- Key Laboratory of chemical Biology and Molecular Engineering Ministry
- Shanxi University
- Institute of Materia Medica
- Chinese Academy of Medical Sciences & Peking Union Medical College
- University of Louisville
- 王迪
- 李科
- 王桂臻
- 李震宇
- 秦雪梅
- 杜冠华
- 张翔
WANG Di- LI Ke
- WANG Gui-zhen
- LI Zhen-yu
- QIN Xue-mei
- DU Guan-hua
- ZHANG Xiang
- Modern Research Center for Traditional Chinese Medicine
- Shanxi University
- College of Chemistry and Chemical Engineering
- Shanxi University
- Key Laboratory of chemical Biology and Molecular Engineering Ministry
- Shanxi University
- Institute of Materia Medica
- Chinese Academy of Medical Sciences & Peking Union Medical College
- University of Louisville