不同分子质量海蚌肝素结构表征及抗凝血与纤溶活性的研究  

Study on structure,anticoagulant and fibrinolytic activities of different molecular weights of heparin from clam Coelomactra antiquata

作　　者：陈观兰 陈菁 陈建平 李瑞 贾学静 刘晓菲 宋兵兵 钟赛意 CHEN Guanlan;CHEN Jing;CHEN Jianping;LI Rui;JIA Xuejing;LIU Xiaofei;SONG Bingbing;ZHONG Saiyi(College of Food Science and Technology,Guangdong Ocean University,Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety,Guangdong Province Engineering Laboratory for Marine Biological Products,Guangdong Provincial Engineering Technology Research Center of Seafood,Zhanjiang 524088,China;Collaborative Innovation Center of Seafood Deep Processing,Dalian Polytechnic University,Dalian 116034,China)

机构地区：[1]广东海洋大学食品科技学院,广东省水产品加工与安全重点实验室,广东省海洋生物制品工程实验室,广东省海洋食品工程技术研究中心,广东湛江524008 [2]大连工业大学海洋食品精深加工关键技术省部共建协同创新中心,辽宁大连116034

出　　处：《食品与发酵工业》2021年第17期119-125,共7页Food and Fermentation Industries

基　　金：国家重点研发计划重点专项(2019YFD0902005);广东省重点领域研发计划项目(2020B1111030004);湛江市科技计划项目(2019A01015)。

摘　　要：为探究不同分子质量海蚌肝素的结构性质及抗凝血与纤溶活性,以海蚌肝素G2及其降解产物DG1、DG2为研究对象,采用高效凝胶色谱测定其分子质量,傅里叶红外光谱、圆二色谱、原子力显微镜、扫描电镜分析结构,体外抗凝血实验及琼脂糖纤溶平板法实验研究体外抗凝血与纤溶活性。结果表明,海蚌肝素G2、DG1、DG2的分子质量分别为60.25 k、24.48 k、6.75 kDa;红外色谱图显示,降解前后海蚌肝素的官能团结构变化不明显,但随着分子质量的降低,官能团或特征峰峰值增加;圆二色谱显示,降解前后海蚌肝素的圆二色谱峰出现在210 nm,但随着分子质量的降低峰值变小;扫描电镜图谱显示,降解后海蚌肝素的碎片状结构及球状结构增加;原子力显微镜图显示,海蚌肝素的网线状结构在降解过程中先被降解成链状结构,后聚集成颗粒结构;DG1的抗Xa/IIa比值最大;不同分子质量海蚌肝素抗凝活性均较为缓和,DG2与G2的抗凝血活性接近,DG2的抗凝活性较低;随着分子质量的降低,纤溶活性升高,DG2的纤溶活性约为普通分子质量肝素的77%,低分子质量肝素的13%。To investigate the structural properties,anticoagulant and fibrinolytic activities of different molecular weights of heparin from clam Coelomactra antiquata,heparin G2 and its two different degradation products,DG1 and DG2,were taken as the research objects in this paper.Their molecular weights were determined by high-performance gel chromatography.Fourier transform infrared spectrometer,circular dichroism spectrum,atomic force microscopy and scanning electron microscopy were used to analyze their structures.Their anticoagulant and fibrinolytic activities were evaluated by extracorporeal anticoagulant assay and fibrinolytic plate assay.The results showed that the average molecular weights of heparin G2,DG1 and DG2 were 60.25 k,24.48 k and 6.75 kDa,respectively.The functional group structure of clam heparin did not change significantly before and after degradation,but the functional group or characteristic peak value increased with the decrease of molecular weight.While the peak value of circular dichroism spectrum of clam heparin decreased.The results of scanning electron microscope showed that the fragmented structure and spherical structure of heparin increased.And atomic force microscope showed that the reticulum linear structure of heparin was degraded into chain structure first,and then aggregated into granular structure.The anticoagulant activity of DG2 was similar to that of G2,but the anticoagulant activity of DG2 was significantly decreased.With the decrease of molecular weight,the fibrinolytic activity increased.

关 键 词：海蚌肝素 分子质量 结构 抗凝血 纤溶活性 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]