不同海洋生物源肝素的理化性质及抗凝血活性

Physicochemical Properties and Anticoagulant Activity of Heparin from Different Marine Organisms

为研究和开发海洋生物源肝素,本实验采用酶解醇沉法从9种海洋生物中提取出粗肝素,采用紫外光谱法、傅里叶变换红外光谱法和高效液相色谱法对其理化性质和结构特性进行分析,并通过体外凝血实验分析其抗凝血活性。结果表明:9种肝素粗品的肝素含量从高到低依次为凡纳滨对虾(Penaeus vannaei)虾头339.92 g/mg、海蚌(Mactra antiquata)239.15 g/mg、泥蚶(Tegillarca granosa)234.85 g/mg、蛏子(Sinonovacula constricta)218.29 g/mg、文蛤(Meretrix meretrix L.)214.78 g/mg、钝缀锦蛤(Tapes dorsatus)212.194 g/mg、海湾扇贝(Argopecten irradias)68.52 g/mg、象拔蚌(Panopea abrupta)61.48 g/mg、菲律宾蛤蜊(Ruditapes philippinarum)24.29 g/mg,其均为吡喃糖,且都含有肝素的主要单糖:氨基葡萄糖、艾杜糖醛酸和葡萄糖醛酸。体外凝血实验结果表明,9种粗肝素效价从高到低依次为海蚌67.1 U/mg、凡纳滨对虾虾头59.3 U/mg、文蛤44.0 U/mg、钝缀锦蛤39.2 U/mg、蛏子35.0 U/mg、泥蚶29.6 U/mg、象拔蚌27.2 U/mg、海湾扇贝27.1 U/mg、菲律宾蛤蜊24.1 U/mg,它们具有不同强度的抗凝血活性,且发挥抗凝血作用的途径不同。实验结果可为海洋源肝素的开发提供参考。

In this study,crude heparin was extracted from 9 marine organisms by enzymatic hydrolysis and then alcohol precipitation.The physicochemical properties and monosaccharide composition of the heparins were analyzed by ultraviolet spectroscopy,Fourier transform infrared spectroscopy and high performance liquid chromatography,and their anticoagulant activities were analyzed by in vitro experiments.The results showed that the heparin contents of the 9 crude heparin samples were 339.92 g/mg for Penaeus vannaei heads,239.15 g/mg for Mactra antiquata,234.85 g/mg for Tegillarca granosa,218.29 g/mg for Sinonovacula constricta,214.78 g/mg for Meretrix meretrix L.,212.194 g/mg for Tapes dorsatus,68.52 g/mg for Argopecten irradias,61.48 g/mg for Panopea abrupta,and 24.29 g/mg for Ruditapes philippinarum.All of them,having pyranose rings,contained glucosamine,iduronic acid,and galacturonic acid.The in vitro experiments showed that the titers of the crude heparin samples were 67.1 U/mg for Mactra antiquata,59.3 U/mg for Penaeus vannaei heads,44.0 U/mg for Meretrix meretrix L.,39.2 U/mg for Tapes dorsatus,35.0 U/mg for Sinonovacula constricta,29.6 U/mg for Tegillarca granosa,27.2 U/mg for Panopea abrupta,27.1 U/mg for Argopecten irradias,and 24.1 U/mg for Ruditapes philippinarum.They exerted different anticoagulant activities through different pathways.This study provides a basis for the development of heparin from marine resources.