厚叶海带岩藻聚糖硫酸酯分离纯化及其体外肿瘤抑制活性研究

彭雍博; 宋悦凡; 武龙; 汪秋宽; 丛海花; 刘舒

doi:10.13386/j.issn1002-0306.2017.17.055

厚叶海带岩藻聚糖硫酸酯分离纯化及其体外肿瘤抑制活性研究

基金项目:

国家自然基金(31471610)；辽宁省海洋与渔业厅渔业推广项目(GCNT-LN-19)；

详细信息

作者简介:
彭雍博 (1992-) , 男, 硕士, 研究方向:农产品加工及贮藏工程, E-mail:pengyongbo2011@126.com。;

汪秋宽 (1962-) , 女, 硕士, 教授, 研究方向:水产品加工及综合利用, E-mail:wqk320@dlou.edu.cn。;

中图分类号: TQ464
计量
- 文章访问数: 182
- HTML全文浏览量: 25
- PDF下载量: 261
出版历程
- 收稿日期: 2017-03-02

Study on separation and purification of fucoidan from Kjellmaniella crassifolia and antitumor activity in vitro

摘要

摘要: 本研究以日本厚叶海带（Kjellmaniella crassifolia）中提取的岩藻聚糖硫酸酯（Fuciodan）为原料,采用DEAESepharose Fast Flow弱阴离子交换柱层析分离纯化,通过苯酚-硫酸法和氯化钡-明胶法检测各组分中多糖及硫酸根（SO₄²）含量,MTT法分析50、200、800μg/m L的日本厚叶海带岩藻聚糖硫酸酯粗品（F）及分离组分对肝癌细胞Hep G2、肺癌细胞A549和结肠癌细胞HT-29细胞存活率影响。研究结果表明,DEAE-Sepharose Fast Flow弱阴离子交换柱对粗品F的分离纯化效果良好,可以得到5个组分（F-0、F-1、F-2、F-3和F-4）;组分F-4多糖与SO₄^2-含量最高,分别为64.25%和26.32%。MTT实验结果显示,岩藻聚糖硫酸酯粗品及各分离组分对三种肿瘤细胞均具有肿瘤抑制活性,不同组分对HepG2、A549、HT-29的最低细胞存活率分别为67.69%、56.03%和27.56%。基于以上研究结果,表明日本厚叶海带岩藻聚糖硫酸酯具有很大的发展潜力,开发成为一种有效的天然抗肿瘤药物。
- 厚叶海带 /
- 岩藻聚糖硫酸酯 /
- 分离纯化 /
- MTT实验 /
- 抗肿瘤
Abstract: In this study, the crude fucoidan extracted from Kjellmaniella crassifolia was taken as raw material, and then purified by DEAE-Sepharose Fast Flow weak anion exchange column. The contents of total polysaccharide and sulfate ( SO₄^2-) of fucoidan in each fraction were determined by phenol-sulfuric acid colorimetry method and barium chloride-gelatin method, respectively.The antitumor activities for all in the crude and separated fractions with 50, 200, 800 μg/m L concentrations against Hep G2 cells, A549 cells and HT-29 cells were determined by MTT assay. The results showed that, crude fucoidan from Kjellmaniella crassifolia could be well separated and purified by DEAE-Sepharose Fast Flow weak anion exchange column, and five different fractions ( F-0, F-1, F-2, F-3, F-4) were obtained. Total polysaccharide and SO₄^2- content in F-4 were the highest, which were 64.25% and 26.32%, respectively. The results of MTT assay indicated that the crude fucoidan and the fractions had significant inhibitory activities on Hep G2, A549 and HT-29 cells.The lowest cell viability of 67.69%, 56.03% and27.56% were obtained for Hep G2, A549 and HT-29 cells respectively.Based on those findings, it suggested that fucoidan from Kjellmaniella crassifolia had great potential to be a kind of effective natural antineoplastic drugs.
- Kjellmaniella crassifolia /
- fucoidan /
- separation and purification /
- MTT assay /
- antitumor

HTML全文

参考文献(29)

[1]	刘舒, 汪秋宽, 何云海, 等.厚叶海带生物活性成分的研究现状[J].水产科学, 2013, 32 (6) :361-367.
[2]	Mak W, Hamid N, Liu T, et al.Fucoidan from New Zealand Undaria pinnatifida:monthly variations and determination of antioxidant activities[J].Carbohydrate polymers, 2013, 95 (1) :606-614.
[3]	Negishi H, Mori M, Mori H, et al.Supplementation of elderly Japanese men and women with fucoidan from seaweed increases immune responses to seasonal influenza vaccination[J].The Journal of nutrition, 2013, 143 (11) :1794-1798.
[4]	Kang S M, Kim K N, Lee S H, et al.Anti-inflammatory activity of polysaccharide purifidy from AMG-assistant extract of Eckionia cava in LPS-stimulated RAW264.7 macrophages[J].Corbohydrate Polymers.2011, 52 (1) :1010-1016.
[5]	Wang J, Zhang Q B, Zhang Z S, et al.In-vitro anticoagulant activity of fucoidan derivatives from brown seaweed Laminaria japonica[J].Chinese Journal of Oceanology&Limnology, 2011, 29 (3) :679-685.
[6]	Ermakova S, Sokolova R, Kim S M, et al.Fucoidans from brown seaweeds Sargassum hornery, Eclonia cava, Costaria costata:structural characteristics and anticancer activity[J].Applied Biochemistry&Biotechnology, 2011, 164 (6) :841-850.
[7]	Ping S, Jia L, Chen X, et al.Structural features and antitumor activity of a purified polysaccharide extracted from Sargassum horneri[J].International Journal of Biological Macromolecules, 2015, 73 (5) :124-130.
[8]	Heim G, O’Doherty J V, O’Shea C J, et al.Maternal supplementation of seaweed-derived polysaccharides improves intestinal health and immune status of suckling piglets[J].Journal of Nutritional Science, 2015, 4 (27) :1-12.
[9]	Lee K W, Jeong D, Na K.Doxorubicin loading fucoidan acetate nanoparticles for immune and chemotherapy in cancer treatment[J].Carbohydrate Polymers, 2013, 94 (2) :850-856.
[10]	史大华, 刘玮炜, 刘永江, 等.低分子量海带岩藻多糖的制备及其抗肿瘤活性研究[J].时珍国医国药, 2012, 23 (1) :53-55.
[11]	李巧梅, 杨丽丽, 王培胜, 等.Fucoidan对Lewis肺癌荷瘤小鼠抑瘤及免疫调节作用[J].中国海洋药物, 2013, 32 (2) :12-16.
[12]	Thinh P D, Menshova R V, Ermakova S P, et al.Structural characteristics and anticancer activity of fucoidan from the brown alga Sargassum mcclurei[J].Marine Drugs, 2013, 11 (5) :1456-1476.
[13]	Ale M T, Meyer A S.Fucoidans from brown seaweeds:an update on structures, extraction techniques and use of enzymes as tools for structural elucidation[J].Rsc Advances, 2013, 3 (22) :8131-8141.
[14]	Mak W, Hamid N, Liu T, et al.Fucoidan from New Zealand Undaria pinnatifida:Monthly variations and determination of antioxidant activities[J].Corbohydrate Polymers, 2013, 95 (1) :606-14.
[15]	刘舒.三种褐藻中岩藻聚糖硫酸酯的纯化及结构分析[D].大连:大连海洋大学, 2013.
[16]	郭峰君, 胡靖, 赵雪.海带岩藻聚糖硫酸酯降解及基本结构分析[J].食品工业科技, 2014, 35 (18) :93-97.
[17]	刘玉凤, 贾淑颖, 刘飞飞, 等.不同取代度的硫酸化肠浒苔多糖抗氧化活性研究[J].食品工业科技, 2016, 37 (19) :142-147, 152.
[18]	李亚娴.羊栖菜多酚粗品的纯化及其抗肿瘤活性研究[D].大连:大连海洋大学, 2016.
[19]	王亚芳, 汪秋宽, 何云海, 等.褐藻多糖硫酸酯与山楂提取物混合剂的降血脂作用[J].大连海洋大学学报, 2015 (3) :309-313.
[20]	Wu L, Sun J, Su X, et al.A review about the development of fucoidan in antitumor activity:Progress and challenges[J].Carbohydrate Polymers, 2016, 154 (8) :96-111.
[21]	Oliveira C, Ferreira A S, Novoa C R, et al.The key role of sulfation and branching on fucoidan antitumor activity[J].Macromolecular Bioscience, 2016:1600340.
[22]	Anastyuk S D, Shevchenko N M, Ermakova S P, et al.Anticancer activity in vitro of a fucoidan from the brown alga Fucus evanescens and its low-molecular fragments, structurally characterized by tandem mass-spectrometry[J].Carbohydrate Polymers, 2012, 87 (1) :186-194.
[23]	You S G.Molecular characteristics of partially hydrolyzed fucoidans from sporophyll of Undaria pinnatifida and their in vitro anticancer activity[J].Food Chemistry, 2010, 119 (2) :554-559.
[24]	Vishchuk O S, Ermakova S P, Zvyagintseva T N.The fucoidans from brown algae of Far-Eastern seas:anti-tumor activity and structure-function relationship.[J].Food Chemistry, 2013, 141 (2) :1211-1217.
[25]	Ale M T, Mikkelsen J D, Meyer A S.Important determinants for fucoidan bioactivity:A critical review of structure-function relations and extraction methods for fucose-containing sulfated polysaccharides from brown seaweeds[J].Marine Drugs, 2011, 9 (10) :2106-2130.
[26]	蔡璐, 王维民, 谌素华, 等.不同组分马尾藻岩藻聚糖硫酸酯对人肺癌细胞A549作用的研究[J].食品工业科技, 2014, 35 (1) :116-119.
[27]	Skriptsova A V, Shevchenko N M, Zvyagintseva T N, et al.Monthly changes in the content and monosaccharide composition of fucoidan from Undaria pinnatifida (Laminariales, Phaeophyta) [J].Journal of Applied Phycology, 2010, 22 (1) :79-86.
[28]	Lee J B, Takeshita A, Hayashi K, et al.Structures and antiviral activities of polysaccharides from Sargassum trichophyllum[J].Carbohydrate Polymers, 2011, 86 (2) :995-999.
[29]	Han Y S, Hee L J, Hun L S.Fucoidan inhibits the migration and proliferation of HT-29 human colon cancer cells via the phosphoinositide-3 kinase/Akt/mechanistic target of rapamycin pathways[J].Molecular Medicine Reports, 2015, 12 (3) :3446-3452.

施引文献

资源附件(0)

计量

文章访问数: 182
HTML全文浏览量: 25
PDF下载量: 261
被引次数: 0

厚叶海带岩藻聚糖硫酸酯分离纯化及其体外肿瘤抑制活性研究

作者简介: 彭雍博 (1992-) , 男, 硕士, 研究方向:农产品加工及贮藏工程, E-mail:pengyongbo2011@126.com。; 汪秋宽 (1962-) , 女, 硕士, 教授, 研究方向:水产品加工及综合利用, E-mail:wqk320@dlou.edu.cn。;

计量

出版历程

Study on separation and purification of fucoidan from Kjellmaniella crassifolia and antitumor activity in vitro

计量

出版历程

目录

作者简介:
彭雍博 (1992-) , 男, 硕士, 研究方向:农产品加工及贮藏工程, E-mail:pengyongbo2011@126.com。;

汪秋宽 (1962-) , 女, 硕士, 教授, 研究方向:水产品加工及综合利用, E-mail:wqk320@dlou.edu.cn。;