小有刺参硫酸软骨素和岩藻聚糖硫酸酯抗血小板聚集活性的比较
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摘要
目的从小有刺参中制备纯化硫酸软骨素和岩藻聚糖硫酸酯及各自的低分子量组分,并比较不同结构硫酸多糖的抗血小板聚集活性,揭示海参硫酸多糖抗血小板聚集活性的构效关系。方法利用离子交换色谱和自由基降解技术,从小有刺参中制备硫酸软骨素和岩藻聚糖硫酸酯及各自的低分子量组分。采用体外凝血酶诱导血小板聚集模型,比较不同结构的硫酸软骨素和岩藻聚糖硫酸酯对血小板聚集率的影响。结果 4～60μg/mL硫酸软骨素和低分子量硫酸软骨素具有显著的抑制血小板聚集的活性,在浓度为20μg/mL时对血小板聚集的抑制率高达94.6%和95.3%,且分子量的降低对其活性无影响。比较不同来源的岩藻聚糖硫酸酯的活性发现,在4～60μg/mL浓度范围内,海参与海带来源的岩藻聚糖硫酸酯都显著抑制血小板聚集,且活性没有显著差异,但经过自由基降解后,海参和海带来源的岩藻聚糖硫酸酯的抗血小板聚集活性都显著降低。结论海参硫酸软骨素的抗血小板聚集活性显著高于岩藻聚糖硫酸酯,海参硫酸多糖可以应用于新型抗血小板聚集药物的研究开发。
Objective The chondroitin sulfate,fucoidan and low-molecular-weight chondroitin sulfate and fucoidan from Holotoria floridona were prepared and their inhibitory effects on the platelet aggregation were compared to reveal the structure-activity relationship of anti-platelet aggregation activity of sulfated polysaccharides from Holotoria floridona.Methods The chondroitin sulfate and fucoidan,and lowmolecular-weight chondroitin sulfate and fucoidan were prepared by anion exchange chromatography and radical oxygen degradation from Holotoria floridona.The anti-platelet aggregation activity was assessed by thrombin-induced platelet aggregation model in vitro.Results 4～60μg/mL chondroitin sulfate and low-molecular-weight chondroitin sulfate exhibited significant higher inhibitory effects on platelet aggregation,their inhibitory rate at 20μg/mL on platelet aggregation reached 94.6% and95.3%,respectively.By comparison,the decrease in molecular weight of chondroitin sulfate did not affect the anti-platelet aggregation activity.The anti-platelet aggregation activities of fucoidans from Holotoria floridona and Laminaria japonica were compared and showed that at 4～60 g/mL,the two fucoidans exhibited similar inhibitory activity on platelet aggregation induced by thrombin.However,the inhibitory activity of low-molecular-weight fucoidans obtained by radical oxygen degradation decreased significantly compared with fucoidans.Conclusion The anti-platelet aggregation activity of chondroitin sulfate and low-molecular-weight chondroitin sulfate were significantly higher than those of fucoidans from Holotoria floridona.Sulfate polysaccharides from sea cucumber had a potential as new anti-platelet drugs.
Objective The chondroitin sulfate,fucoidan and low-molecular-weight chondroitin sulfate and fucoidan from Holotoria floridona were prepared and their inhibitory effects on the platelet aggregation were compared to reveal the structure-activity relationship of anti-platelet aggregation activity of sulfated polysaccharides from Holotoria floridona.Methods The chondroitin sulfate and fucoidan,and lowmolecular-weight chondroitin sulfate and fucoidan were prepared by anion exchange chromatography and radical oxygen degradation from Holotoria floridona.The anti-platelet aggregation activity was assessed by thrombin-induced platelet aggregation model in vitro.Results 4～60μg/mL chondroitin sulfate and low-molecular-weight chondroitin sulfate exhibited significant higher inhibitory effects on platelet aggregation,their inhibitory rate at 20μg/mL on platelet aggregation reached 94.6% and95.3%,respectively.By comparison,the decrease in molecular weight of chondroitin sulfate did not affect the anti-platelet aggregation activity.The anti-platelet aggregation activities of fucoidans from Holotoria floridona and Laminaria japonica were compared and showed that at 4～60 g/mL,the two fucoidans exhibited similar inhibitory activity on platelet aggregation induced by thrombin.However,the inhibitory activity of low-molecular-weight fucoidans obtained by radical oxygen degradation decreased significantly compared with fucoidans.Conclusion The anti-platelet aggregation activity of chondroitin sulfate and low-molecular-weight chondroitin sulfate were significantly higher than those of fucoidans from Holotoria floridona.Sulfate polysaccharides from sea cucumber had a potential as new anti-platelet drugs.
引文
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[11]Mour2o P A,Bastos I G.Highly acidic glycans from sea cucumbers.Isolation and fractionation of fucose-rich sulfated polysaccharides from the body wall of Ludwigothureagrisea[J].Eur J Biochem,1987,166(3):639-645.
[12]赵珊,许加超,付晓婷,等.褐藻寡糖分子量测定方法的研究[J].食品工业科技,2011,32(12):486-488.
[13]Strydom D J.Chromatographic separation of 1-phenyl-3-methy-5pyrazolone-derivatized neutral,acidic and basic aldoses[J].J Chromatogr A,1994,678(1):17-23.
[14]Fu D T,O'Neill R A.Monosaccharide composition analysis of oligosaccharides and glycoproteins by high-performance liquid chromatography[J].Anal Biochem,1995,227(2):377-384.
[15]尹利昂,陈士国,薛长湖,等.4种海参中含岩藻糖支链的硫酸软骨素化学组成差异的分析[J].中国海洋大学学报(自然科学版),2009,39(s1):63-68.
[16]Harrison P.Platelet function analysis[J].Blood Rev,2005,19(2):111-123.
[2]李晓燕,许琳,谈红.抗凝与溶栓[M].北京:科学技术文献出版社,2011:28-30.
[3]丛悦,柳晓兰,余祖胤,等.知母皂苷抑制血小板聚集的活性成分筛选及构效关系分析[J].解放军医学杂志,2010,35(11):1370-1373.
[4]Wu M Y,Xu S M,Zhao J H,et al.Physicochemical characteristics and anticoagulant activities of low molecular weight fractions by free-radical depolymerization of a fucosylated chondroitin sulphate from sea cucumber Thelenataananas[J].Food Chem,2010,122(3):716-723.
[5]Fonseca R J,Santos G R,Mour2o P A.Effects of polysaccharides enriched in 2,4-disulfated fucose units on coagulation,thrombosis and bleeding[J].Thromb Haemost,2009,102(5):829-836.
[6]胡靖,李八方,谢洁玲,等.北极海参硫酸软骨素及其低分子量糖抗血栓活性研究[J].中国海洋药物,2016,35(6):46-51.
[7]刘静文,史姣霞,谢洁玲,等.海带低分子量岩藻聚糖硫酸酯的制备和抗血小板聚集活性研究[J].中国海洋药物,2017,36(1):19-26.
[8]王晶,张全斌.褐藻多糖硫酸酯的结构与生物活性研究[J].海洋科学集刊,2017(52):68-89.
[9]Azevedo T C,Bezerra M E,Santos M G,et al.Heparinoids algal and their anticoagulant,hemorrhagic activities and platelet aggregation[J].Biomed Pharmacother,2009,63(7):477-483.
[10]Zhao X,Dong S Z,Wang J F,et al.A comparative study of antithrombotic and antiplatelet activities of different fucoidans from Laminariajaponica[J].Thromb Res,2012,129(6):771-778.
[11]Mour2o P A,Bastos I G.Highly acidic glycans from sea cucumbers.Isolation and fractionation of fucose-rich sulfated polysaccharides from the body wall of Ludwigothureagrisea[J].Eur J Biochem,1987,166(3):639-645.
[12]赵珊,许加超,付晓婷,等.褐藻寡糖分子量测定方法的研究[J].食品工业科技,2011,32(12):486-488.
[13]Strydom D J.Chromatographic separation of 1-phenyl-3-methy-5pyrazolone-derivatized neutral,acidic and basic aldoses[J].J Chromatogr A,1994,678(1):17-23.
[14]Fu D T,O'Neill R A.Monosaccharide composition analysis of oligosaccharides and glycoproteins by high-performance liquid chromatography[J].Anal Biochem,1995,227(2):377-384.
[15]尹利昂,陈士国,薛长湖,等.4种海参中含岩藻糖支链的硫酸软骨素化学组成差异的分析[J].中国海洋大学学报(自然科学版),2009,39(s1):63-68.
[16]Harrison P.Platelet function analysis[J].Blood Rev,2005,19(2):111-123.