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来自长枝蜈蚣藻的硫酸化多糖的纯化、部分特性鉴定及生物活性

Purification, partial characterization and bioactivity of sulfated polysaccharides from Grateloupia livida.

作者信息

Tang Liu, Chen Yicun, Jiang Zebin, Zhong Shuping, Chen Weizhou, Zheng Fuchun, Shi Ganggang

机构信息

Department of Pharmacy, First Affiliated Hospital, Shantou University Medical College, Shantou 515041, China.

Department of Pharmacology, Shantou University Medical College, Shantou 515041, China.

出版信息

Int J Biol Macromol. 2017 Jan;94(Pt A):642-652. doi: 10.1016/j.ijbiomac.2016.10.067. Epub 2016 Oct 20.

Abstract

Purification, preliminary characterization and bioactivity of polysaccharides from Grateloupia livida (GL) were investigated. Three water-soluble sulfated polysaccharide fractions (GLP-1, GLP-2 and GLP-3) were isolated and purified from the edible and medicinal red seaweed, Grateloupia livida (Harv.) Yamada by DEAE Sepharose CL-6B and Sephadex G-100 column chromatography, and chemical characterization was performed by HPGPC, GC-MS, FT-IR and SEM. In addition, anticoagulant activities were determined by activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) using normal human plasma in vitro. The antioxidant activities against DPPH and ABTS radicals were evaluated and compared. The molecular weights of GLP-1, GLP-2 and GLP-3 were 39.5, 60.4 and 3.36kDa, respectively. Monosaccharide analysis revealed that three polysaccharide fractions were homopolysaccharides and comprised of galactose only. Anticoagulant assays indicated that crude GLP, and purified GLP-1 and GLP-2 effectively prolonged APTT and TT, but not PT. All polysaccharide fractions exhibited significant in vitro antioxidant activities in a dose-dependent manner. GLP-2 showed consistently better anticoagulant and antioxidant activities compared with GLP, GLP-1 and GLP-3. These results demonstrate that sulfated polysaccharides isolated from Grateloupia livida can serve as readily available alternative natural sources of anticoagulant and antioxidant agents.

摘要

对蜈蚣藻(GL)多糖的纯化、初步表征及生物活性进行了研究。通过DEAE Sepharose CL - 6B和Sephadex G - 100柱色谱法,从食用和药用红藻蜈蚣藻(Grateloupia livida (Harv.) Yamada)中分离并纯化出三种水溶性硫酸化多糖组分(GLP - 1、GLP - 2和GLP - 3),并采用高效凝胶渗透色谱法(HPGPC)、气相色谱 - 质谱联用仪(GC - MS)、傅里叶变换红外光谱仪(FT - IR)和扫描电子显微镜(SEM)进行化学表征。此外,使用正常人血浆在体外通过活化部分凝血活酶时间(APTT)、凝血酶时间(TT)和凝血酶原时间(PT)测定抗凝活性。评估并比较了对DPPH和ABTS自由基的抗氧化活性。GLP - 1、GLP - 2和GLP - 3的分子量分别为39.5、60.4和3.36 kDa。单糖分析表明,三种多糖组分均为同多糖,仅由半乳糖组成。抗凝试验表明,粗制GLP以及纯化后的GLP - 1和GLP - 2能有效延长APTT和TT,但不影响PT。所有多糖组分在体外均呈现出显著的剂量依赖性抗氧化活性。与GLP、GLP - 1和GLP - 3相比,GLP - 2始终表现出更好的抗凝和抗氧化活性。这些结果表明,从蜈蚣藻中分离出的硫酸化多糖可作为现成的抗凝剂和抗氧化剂的替代天然来源。

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