低聚κ-卡拉胶硫酸酯的制备及其抗凝血活性研究  

Preparation of sulfated low molecular weight κ-carrageenan and study of its anticoagulant activity

作　　者：任永峰 辛萌[1,2,3] 李帆 李建杰[1,2,3] 王箴言 胡婷 李春霞[1,2,3] REN Yongfeng;XIN Meng;LI Fan;LI Jianjie;WANG Zhenyan;HU Ting;LI Chunxia(Key Laboratory of Marine Drugs,Ministry of Education,Shandong Key Laboratory of Glycoscience and Glycoengineering,School of Medicine and Pharmacy,Ocean University of China,Qingdao 266003,China;Laboratory for Marine Drugs and Bioproducts,Qingdao Marine Science and Technology Center,Qingdao 266237,China;Marine Biomedical Research Institute of Qingdao,Qingdao 266100,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]青岛海洋科技中心海洋药物与生物制品功能实验室,山东青岛266237 [3]青岛海洋生物医药研究院,山东青岛266100

出　　处：《中国海洋药物》2024年第5期9-16,共8页Chinese Journal of Marine Drugs

基　　金：山东省泰山产业领军人才工程蓝色人才专项;山东省重大科技创新工程专项(2018SDKJ0404-3)资助。

摘　　要：目的 制备低聚κ-卡拉胶硫酸酯(LCS),并对其抗凝血活性进行研究。方法 以κ-卡拉胶为原料,通过硫酸降解制备得低聚κ-卡拉胶(LC),进一步通过三氧化硫吡啶法磺化制备得LCS。采用体外抗凝血实验,检测LCS对大鼠血浆和绵羊血浆活化部分凝血活酶时间(APTT)和凝血酶时间(TT)的影响。采用正常小鼠,皮下注射给药1 h后取血,检测LCS对小鼠血浆凝血四项APTT、TT、凝血酶原时间(PT)和纤维蛋白原(FIB)含量的影响,研究其体内抗凝血活性。此外,采用小鼠尾部出血实验评估其潜在的出血副作用。结果 制备的LCS重均分子量为6.6 kDa,硫酸根含量为48.9%,其具有显著的抗凝血作用。体外实验结果表明,在大鼠血浆和绵羊血浆中,LCS均可以显著延长APTT,在绵羊血浆中LCS可以延长TT,但在大鼠血浆中对TT的延长效果不明显,存在种属差异性。体内实验结果表明,LCS在小鼠体内可以显著延长APTT,但对TT、PT和FIB的作用效果不明显。小鼠尾部出血实验结果表明,在与依诺肝素钠等效剂量下,LCS对出血时间无显著影响,高剂量下LCS造成的出血风险低于依诺肝素钠。结论 LCS具有良好的抗凝血活性和低出血风险,主要是作用于内源性凝血途径。Objective Preparation of sulfated low molecular weight κ-carrageenan(LCS) and study of its anticoagulant activity.Methods Using κ-carrageenan as the raw material,LCS was prepared by sulfuric acid degradation and sulfated by trioxopyridine.The effects of LCS on activated partial thrombin time(APTT)and thrombin time(TT) in rat and sheep plasma were determined in vitro.Blood was collected one hour after subcutaneous injection in mice.APTT,TT,prothrombin time(PT),and fibrinogen(FIB) content were used to evaluate the anticoagulant effect of LCS.In addition,the tail bleeding test was used to evaluate the potential bleeding side effects.Results The weight-average molecular weight of LCS was 6.6 kDa,the sulfate content was 48.9%,and it had a significant anticoagulant effect.In vitro results showed that LCS could significantly prolong APTT in rat and sheep plasma.LCS could prolong TT in sheep plasma,but not significantly in rat plasma,indicating the species difference.In vivo results showed that LCS could significantly prolong APTT in mice,but had an insignificant effect on TT,PT,and FIB.The results of tail bleeding in mice showed that LCS had an insignificant effect on bleeding time at the equivalent dose of enoxaparin sodium.And the risk of bleeding at higher doses of LCS was lower than that of enoxaparin sodium.Conclusion LCS has good anticoagulant activity with a low bleeding risk,mainly through the intrinsic coagulation pathway.

关 键 词：κ-卡拉胶硫酸酯 抗凝血活性 内源性凝血途径 

分 类 号：R931.77[医药卫生—生药学]