2种颜色仿刺参Apostichopus japonicus中糖胺聚糖表达谱的差异性研究  

Differences of glycosaminoglycans expression profiles of Apostichopus japonicus in two colors

作　　者：韩青 周涵 王维雯 黄曦玥 陈刚 陈四清[4] 于广利[1,2] 李国云 HAN Qing;ZHOU Han;WANG Wei-wen;HUANG Xi-yue;CHEN Gang;CHEN Si-qing;YU Guang-li;LI Guo-yun(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 for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology,Qingdao 266237,China;Qingdao Jinhai Fuyuan Ocean Industry Co.,Ltd,Qingdao 266075,China;Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,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 [3]青岛金海富源海洋实业有限公司,山东青岛266075 [4]中国水产科学研究院黄海水产研究所,山东青岛266071

出　　处：《中国海洋药物》2023年第2期1-9,共9页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金项目(31971210,81991522);泰山学者攀登计划(TSPD20210304)资助。

摘　　要：目的基于液质联用和糖组学分析技术,比较2种颜色仿刺参中的糖胺聚糖(GAGs)组成,获得2种颜色仿刺参的糖胺聚糖表达谱,从而完善海参糖链表达谱信息,促进无脊椎动物的糖链组成研究。方法对2种颜色仿刺参进行脱脂和蛋白酶酶解处理,采用离子交换柱纯化以去除蛋白和高硫酸化聚糖等杂质并获得糖胺聚糖。采用GAGs酶解获得GAGs二糖并对其进行2-AMAC荧光标记,基于RP-C18-MS/MS-MRM方法进行GAGs表达谱差异分析。结果在2种颜色仿刺参中CS含量均最高,为2.5~4.5μg/g,HA含量次之,HS的含量最低。海参中GAGs总量远低于海参中岩藻糖基化硫酸软骨素和岩藻聚糖硫酸酯的含量,白色仿刺参的GAGs总量明显高于青色仿刺参。2种仿刺参中CS二糖以CS-0S、CS-4S、CS-6S为主,HS二糖主要是HS-NS6S和HS-TriS,但是两者在GAGs二糖含量、质量分数、硫酸化模式和硫酸化程度上均存在较大差异。结论首次获得2种颜色仿刺参糖胺聚糖表达谱信息,有助于了解2种颜色仿刺参来源内源性糖链结构信息,为海参活性成分的研究开发提供依据。Objective Based on liquid chromatography-mass spectrometry and glycomic analysis techniques,the expression profiles of glycosaminoglycans of Apostichopus japonicus in different colors were analyzed and compared.Glycosaminoglycans expression information of Apostichopus japonicus in different colors was obtained so as to improve the glycans expression information of sea cucumber and promote the glycans research of invertebrates.Methods First,the two colors Apostichopus japonicus were degreased and treated by Pronase,and then purified by ion exchange column to remove protein and sulfated polysaccharides and obtain glycosaminoglycans.GAGs disaccharides were obtained by GAGs enzymesSand labeled with 2-AMAC fluorescence,and then the difference of GAGs expression profiles were analyzed based on RP-C18-MS/MS-MRM method.Results CS was the highest content in the range of 2.5~4.5μg/g,followed by HA,HS was the lowest content in two colors Apostichopus japonicus.The total amount of GAGs in sea cucumber was much lower than the contents of fucosylated chondroitin sulfate and Fucoidan in sea cucumber,the total amount of GAGs in white Apostichopus japonicus was significantly higher than that in green Apostichopus japonicus.CS-0S,CS-4S and CS-6S were the main CS disaccharides in the two kinds of Apostichopus japonicus,while HS-NS6S and HS-TriS were the main HS disaccharides.However,there were significant differences between two colors Apostichopus japonicus in disaccharides content,percentage content,sulfated modification mode and the degree of sulfated modification of GAGs.Conclusion In this paper,the glycosaminoglycans expression profiles of Apostichopus japonicus in different colors were obtained for the first time,which was helpful to understand the endogenous glycans information of Apostichopus japonicus in different colors and provide basis for the study of active ingredients of sea cucumber.

关 键 词：液质联用技术 糖胺聚糖 仿刺参 差异性比较 

分 类 号：R917[医药卫生—药物分析学]