球参多糖的分离纯化及体外抗凝血活性研究  

Isolation,purification,and anticoagulant analysis of polysac-charides from sea cucumber Phyllophorus proteus

作　　者：刘静雯 吴宁[1,3] 耿丽华[1,3] 王晶[1,3] 岳洋 张全斌[1,3] LIU Jingwen;WU Ning;GENG Lihua;WANG Jing;YUE Yang;ZHANG Quanbin(Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China;University of Chinese Academy of Sciences,Beijing 100049,China;Center for Ocean Mega-Science,Chinese Academy of Sciences,Qingdao 266071,China)

机构地区：[1]中国科学院海洋研究所实验海洋生物学重点实验室,山东青岛266071 [2]中国科学院大学,北京100049 [3]中国科学院海洋大科学研究中心,山东青岛266071

出　　处：《海洋科学》2024年第9期34-43,共10页Marine Sciences

基　　金：福建省STS计划配套项目(2023T3057)。

摘　　要：球参(Phyllophorus proteus)是一种产于印度尼西亚等热带海域,具有较大开发利用潜力的低价值海参。本研究利用酶解碱解联用与离子交换柱层析相结合的方法,从球参中分离到2种硫酸化多糖,经理化性质分析,确定2种多糖分别为岩藻聚糖硫酸酯(FS)与岩藻糖化糖胺聚糖(FG)。分别采用Sephadex G-100凝胶柱层析对2种多糖进行纯化,获得了2种多糖纯化组分,之后采用稀酸水解FS得到dFS,采用β-消除降解FG获得dFG。体外抗凝血活性实验结果显示FG能够显著延长活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)和纤维蛋白原(FIB)时间,表明其具有非常显著的抗凝血活性。dFG能够显著延长APTT和FIB的凝血时间,但对PT没有明显影响,即dFG可以影响内源性凝血途径,但不能影响外源性凝血途径,因此调节FG的分子量可以加强对特定凝血途径的选择性作用,从而在保持较好的抗凝血活性的同时降低出血风险,以上结果提示FG及dFG均具有开发成为抗凝血剂的潜力。In this study,two sulfated polysaccharides were isolated from Phyllophorus proteus using enzymat-ic–alkaline coupling and Sepharose DEAE Fast Flow column chromatography,and Sephadex G100 column chro-matography was used for further purification.Physicochemical property analysis indicated that the two sea cucum-ber polysaccharides were fucan sulfate(FS)and fucosylated glycosaminoglycan(FG).Two low molecular weight polysaccharides—dFS and dFG—were obtained through the degradation of FS using 10 mmol/L TFA and degrada-tion of FG via theβ-elimination method,respectively.The in vitro anticoagulant assay demonstrated that FG showed good anticoagulant activity and significantly prolonged the clotting times of activated partial thromboplas-tin time(APTT),prothrombin time(PT),thrombin time(TT),and FIB.dFG significantly prolonged the clotting times of APTT,TT,and FIB but showed no significant effect on the clotting time of PT.Therefore,adjusting the molecular weight of FG can enhance the selective effects on specific coagulation pathways,thus reducing the risk of bleeding while maintaining optimal anticoagulant activities.These results indicated that FG and dFG can be de-veloped as anticoagulants.

关 键 词：球参 岩藻聚糖硫酸酯 岩藻糖化糖胺聚糖 抗凝血 

分 类 号：R284.2[医药卫生—中药学]