| 王斐,杨鲁瑶,李淇,等.多叶鳃海麒麟Placobranchus ocellatus多糖的提取、理化性质及抗凝活性研究[J].中国海洋药物,2025,44(6):1-7. |
| 多叶鳃海麒麟Placobranchus ocellatus多糖的提取、理化性质及抗凝活性研究 |
| Extraction, physicochemical property and anticoagulant activity of polysaccharides from Placobranchus ocellatus |
| 投稿时间：2024-04-22 修订日期：2024-06-14 |
| DOI：10.13400/j.cnki.cjmd.2025.06.001 |
| 中文关键词: 多叶鳃海麒麟 多糖 结构表征 O-糖链 抗凝活性 |
| English Keywords:Placobranchus ocellatus polysaccharides structural characterization O-glycans anticoagulant activity |
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| 中文摘要: |
| 目的 采用酶解法从多叶鳃海麒麟(Placobranchus ocellatus)中提取多糖,对其理化性质和结构进行分析,并初步评价其抗凝活性。方法 通过采用木瓜蛋白酶解提取多叶鳃海麒麟多糖(PO-E)；采用硫酸苯酚法、硫酸钡比浊法、PMP柱前衍生高效液相色谱法、高效凝胶色谱法分别测定其总糖含量、硫酸根含量、单糖组成、相对分子质量,采用核磁共振波谱法(NMR)对其结构特征进行初步表征；采用液质联用分析(PGC-ESI-MS/MS)方法对PO-E中的N, O-糖链进行鉴定分析；通过测定活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)评价PO-E的体外抗凝血活性。结果 采用酶法提取获得的多叶鳃海麒麟多糖PO-E主要为硫酸化半乳聚糖,并含有一定的糖蛋白,通过液质联用分析鉴定发现,PO-E中存在7种O-糖链,糖链结构类型主要以Core 2和Core 1形式存在。体外抗凝活性评价结果显示PO-E对APTT具有延长作用,对PT、TT无延长作用,这表明其通过内源性凝血途径发挥抗凝活性。结论 酶法提取的多叶鳃海麒麟多糖PO-E为含有O-糖基修饰的硫酸化半乳聚糖,具有一定的抗凝活性,为相关海洋抗凝药物的开发提供物质基础与理论依据。 |
| English Summary: |
| Objective The polysaccharide was extracted from Placobranchus ocellatus by enzymatic extraction, and its physicochemical properties, structural characteristics, and anticoagulant activities were investigated. Methods The polysaccharide derived from Placobranchus ocellatus (PO-E) was obtained by papain degradation. The phenol-sulfuric acid method, barium sulfate turbidimetry, pre-column derivatization high performance liquid chromatography, and high performance gel permeation chromatography were adopted to determine the total polysaccharide content, sulfate content, monosaccharide composition, and molecular weight, respectively. The chemical structure of PO-E was preliminary characterized by nuclear magnetic resonance spectroscopy (NMR). The N, O-glycans was analyzed by liquid mass analysis (PGC-ESI-MS/MS). The anticoagulant activity in vitro of PO-E was evaluated by determining activated partial thromboplastin time (APTT), thrombin time (PT), and prothrombin time (TT). Results PO-E was mainly sulfate galactans and it had glycoproteins. Seven O-glycans were identified in PO-E, and structural types of O-glycans were mainly Core 2 and Core 1. PO-E could prolong APTT in vitro but can’t prolong PT and TT, which indicated it exerted anticoagulant activity through intrinsic coagulation pathway. Conclusion The polysaccharide extracted from Placobranchus ocellatus (PO-E) by enzymatic extraction was sulfated galactans containting O-glycosylation, and it had anticoagulant activity. It provides material basis and theoretical basis for the development of marine anticoagulant drugs. |
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