盾果藻Carpopeltis affinis多糖的提取分离、理化性质及抗病毒活性研究  

Extraction,Separation,Physicochemical Property and Antiviral Activity of polysaccharides from Carpopeltis affinis

作　　者：孙继芹 牛庆凤 秦吉 张烜 艾旭泽 于广利[1,2] 王伟 李国云[1,2] SUN Ji-qn;NIU Qing-feng;QIN Ji;ZHANG Xuan;AI Xu-ze;YU Guang-li;WANG Wei;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 Municipal Bureau of Ecology and Environment,Shinan Branch,Qingdao 266003,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]青岛市生态环境局市南分局,山东青岛266003

出　　处：《中国海洋药物》2023年第2期10-16,共7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：目的采用不同方法提取盾果藻Carpopeltis affinis中多糖,对其理化性质和结构进行分析,并初步评价其抗病毒活性。方法依次通过冷水、热水(80℃)和2%NaOH溶液提取盾果藻多糖,通过硫酸苯酚、离子色谱法、PMP柱前衍生高效液相色谱法等对其总糖、硫酸根含量、单糖组成等进行分析,利用核磁共振波谱技术初步探究其结构。通过细胞病变效应(CPE)和假病毒抑制实验初步探究其抗病毒活性。结果3种多糖冷水提取(CCAP)、热水提取(HCAP)、2%NaOH提取(ACAP)均主要含硫酸半乳糖,热水浸提的HCAP得率及纯度较高,且主要为ι-卡拉胶。抗病毒活性研究表明HCAP对流感病毒、疱疹病毒和新冠假病毒具有显著的抑制作用,其中对H1N1株Influenza(A/Virginia/ATCC1/2009)的抑制作用最为显著(半抑制浓度IC50=0.03μg/mL,安全指数SI>66666.6)。结论热水提取获得的盾果藻多糖为ι-卡拉胶,其对多种病毒均具有显著的抑制作用,该结果丰富了海洋糖库,为海洋抗病毒药物的开发提供了理论基础。Objective To extract polysaccharides from Carpopeltis affinis by different methods,investigate their physicochemical properties,structural characteristics and antiviral activities.Methods Three polysaccharides were prepared from a red alga Carpopeltis affinis by sequential extraction with cold water,hot water at 80℃and 2%sodium hydroxide water solution.The total sugar,sulfate content and monosaccharide composition of three polysaccharides were determined by phenol-sulfuric acid,ion chromatography and pre-column derivatization high performance liquid chromatography,respectively.The structure of polysaccharides was characterized by nuclear magnetic resonance spectroscopy.Cytopathic effect(CPE)and pseudovirus inhibition assay were used to preliminary investigate their antiviral activities.Results Three polysaccharides CCAP(cold water),HCAP(hot water at 80℃),ACAP(2%sodium hydroxide water solution at 60℃)were all mainly composed of sulfated galactans.The yield and purity of HCAP was higher and the NMR data indicated it wasτ-carrageenan.The antiviral activity studies have proved that HCAP could inhibit influenza A virus,herpes virus and sever acute respiratory syndrome coronavirus 2 pseudovirus.Among them,the inhibition effect on H1N1 strain Influenza(A/Virginia/ATCC1/2009)was the most significant(IC50=0.03μg.mL-1,SI>66666.6).Conclusion The hot water extracted fraction was mainlyτ-carrageenan and it had a significant inhibitory effect on several kinds of virus.The results enrich the marine carbohydrate library.Meanwhile,it provides the basis for the development of marine antiviral drugs.

关 键 词：盾果藻 多糖 结构表征 抗病毒活性 

分 类 号：R931[医药卫生—生药学]