Chen Shiguo, Li Guoyun, Wu Nian, Guo Xin, Liao Ningbo, Ye Xingqian, Liu Donghong, Xue Changhu, Chai Wengang
College of Biosystem Engineering and Food Science, Zhejiang University, Hangzhou 310029, China.
Biochim Biophys Acta. 2013 Apr;1830(4):3054-66. doi: 10.1016/j.bbagen.2013.01.001. Epub 2013 Jan 9.
The aim is to compare the structures, anticoagulant and antithrombotic activities of two fucosylated chondroitin sulfates isolated from sea cucumbers Isostichopus badionotus (fCS-Ib) and Pearsonothuria graeffei (fCS-Pg), which were known to have different sulfation patterns on the fucose branches.
The structures of fCSs were identified using 2D NMR. Anticoagulant activities were measured by activated partial thromboplastin time (APTT) and thrombin time (TT), and inhibition of factors IIa, Xa and XIIa was assessed in vitro. Antithrombotic activity was determined ex vivo by measuring the length and weight of the thrombus generated.
The two fCSs had identical chondroitin sulfate E backbones and similar fucose branches, but different sulfation patterns of the fucose branches. The fucose branch in fCS-Ib was mainly 2,4-O-sulfated whereas that in fCS-Pg was mainly 3,4-O-sulfated. In vitro assay indicated that fCS-Pg possessed much lower potency than fCS-Ib in prolonging APTT/TT and in inhibition of thrombin. However, they both exhibited similar inhibitory effects on factor X activation by intrinsic tenase complex, and on thrombus generation. Furthermore, both fCSs significantly activated factor XII, which has been proved to be associated with adverse clinical events associated with heparin contaminated by oversulfated chondroitin sulfate.
The 2,4-O-sulfated fucose branch is the key structural factor of fCSs for prolonged APTT/TT and inhibition of thrombin, whereas the inhibitory effect of fCSs on factor X, XII activation and thrombus generation was attributed to the overall structure of fCS polysaccharide. GENERAL IMPORTANCE: Both fCSs have well defined structures and can be readily quality-controlled, and therefore may be potential alternatives for heparin as anticoagulant and antithrombotic drugs.
目的是比较从海参伊斯托刺参（fCS-Ib）和格氏海参（fCS-Pg）中分离得到的两种岩藻糖基化硫酸软骨素的结构、抗凝和抗血栓活性，已知这两种硫酸软骨素在岩藻糖支链上具有不同的硫酸化模式。
使用二维核磁共振（2D NMR）鉴定硫酸软骨素的结构。通过活化部分凝血活酶时间（APTT）和凝血酶时间（TT）测定抗凝活性，并在体外评估对因子IIa、Xa和XIIa的抑制作用。通过测量生成血栓的长度和重量在体外测定抗血栓活性。
两种硫酸软骨素具有相同的硫酸软骨素E主链和相似的岩藻糖支链，但岩藻糖支链的硫酸化模式不同。fCS-Ib中的岩藻糖支链主要是2,4-O-硫酸化的，而fCS-Pg中的岩藻糖支链主要是3,4-O-硫酸化的。体外试验表明，fCS-Pg在延长APTT/TT和抑制凝血酶方面的效力远低于fCS-Ib。然而，它们对由内源性凝血酶原酶复合物激活因子X以及对血栓生成均表现出相似的抑制作用。此外，两种硫酸软骨素均能显著激活因子XII，这已被证明与因硫酸化过度的硫酸软骨素污染肝素而导致的不良临床事件有关。
2,4-O-硫酸化的岩藻糖支链是硫酸软骨素延长APTT/TT和抑制凝血酶的关键结构因素，而硫酸软骨素对因子X、XII激活和血栓生成的抑制作用归因于硫酸软骨素多糖的整体结构。一般重要性：两种硫酸软骨素均具有明确的结构且易于质量控制，因此可能是肝素作为抗凝和抗血栓药物的潜在替代品。
