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从狗鲨软骨中分离、纯化硫酸软骨素及其结构特征:体外抗凝血和抗增殖活性。

Isolation, Purification and Structural Characterestics of Chondroitin Sulfate from Smooth hound Cartilage: In vitro Anticoagulant and Antiproliferative Properties.

机构信息

Laboratory of Plant Improvement and Valorization of Agroressources, National School of Engineering of Sfax (ENIS), University of Sfax, Sfax 3038, Tunisia.

Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.

出版信息

Carbohydr Polym. 2018 Oct 1;197:451-459. doi: 10.1016/j.carbpol.2018.06.040. Epub 2018 Jun 8.

DOI:10.1016/j.carbpol.2018.06.040
PMID:30007634
Abstract

Chondroitin sulfate was extracted from the cartilage of smooth hound (CSSH) and then purified by anion exchange chromatography. The structual characteristic of CSSH was evaluated by acetate cellulose electrophoresis, FTIR, C NMR and SAX-HPLC. Molecular weight of CSSH was average 68.78 KDa. Disaccharide analysis indicated that CSSH was predominately composed of monosulfated disaccharides in position 6 and 4 of the N-acetylgalactosamine (45.34% and 32.49%, respectively). CSSH was tested for in vitro anticoagulant activity using the three classical coagulation assays (activated partial thromboplastin time (aPTT), prothrombine time (TT) and thrombin time (PT) tests). The finding showed that CSSH prolonged significatively (p < 0.05), aPTT, TT and PT about 1.4, 3.44 and 1.21 fold, respectively, greater than that of the negative control at a concentration of 100 μg/ml. The CSSH caused a significant antiproliferative activity against HCT116 cell, which was 79% of cell proliferation inhibition at the concentration of 1000 μg/ml. Further, CSSH presented no toxicity against the normal cells and no hemolysis towards bovine erythrocytes for all concentrations tested. CSSH demonstrated hopeful antiproliferative and anticoagulant potential, which may be used as a novel and effective drug.

摘要

硫酸软骨素从灰星鲨软骨中提取出来,然后通过阴离子交换层析进行纯化。采用醋酸纤维素电泳、FTIR、C NMR 和 SAX-HPLC 对 CSSH 的结构特征进行了评估。CSSH 的平均分子量为 68.78 kDa。二糖分析表明,CSSH 主要由 N-乙酰半乳糖胺 6 位和 4 位的单硫酸盐二糖组成(分别为 45.34%和 32.49%)。采用三种经典凝血测定法(活化部分凝血活酶时间(aPTT)、凝血酶原时间(TT)和凝血时间(PT)试验)检测 CSSH 的体外抗凝活性。结果表明,在 100μg/ml 浓度下,CSSH 分别显著延长(p<0.05)aPTT、TT 和 PT 约 1.4、3.44 和 1.21 倍,比阴性对照长。CSSH 对 HCT116 细胞表现出显著的增殖抑制活性,在 1000μg/ml 浓度下,细胞增殖抑制率为 79%。此外,CSSH 对正常细胞无毒性,对所有测试浓度的牛红细胞无溶血作用。CSSH 显示出有希望的抗增殖和抗凝活性,可能被用作一种新型有效的药物。

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