铜藻褐藻聚糖硫酸酯的分离纯化、结构组成及保肝护肝作用

卓丹琪, 张雪楠, 李雨晴, 刘诗萌, 任丹丹, 何云海, 汪秋宽

大连海洋大学学报 ›› 2022, Vol. 37 ›› Issue (2) : 295-303.

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大连海洋大学学报 ›› 2022, Vol. 37 ›› Issue (2) : 295-303. DOI: 10.16535/j.cnki.dlhyxb.2021-304

铜藻褐藻聚糖硫酸酯的分离纯化、结构组成及保肝护肝作用

  • 卓丹琪,张雪楠,李雨晴,刘诗萌,任丹丹,何云海,汪秋宽
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Purification,structural characteristics and hepatoprotective effects of fucoidan from seaweed Sargassum horneri

  • ZHUO Danqi, ZHANG Xuenan, LI Yuqing, LIU Shimeng, REN Dandan, HE Yunhai, WANG Qiukuan
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摘要

为研究铜藻褐藻聚糖硫酸酯的结构组成及保肝护肝作用,以铜藻Sargassum horneri褐藻聚糖硫酸酯(fucoidan)粗品为原料,经DEAE-Sepharose Fast Flow阴离子交换柱分离纯化,采用高效液相色谱及红外光谱对粗多糖(TF)及其纯化组分的单糖组成和结构特征进行分析,并建立CCl4诱导的肝脏损伤动物模型,进行各组分的保肝作用试验。结果表明:从铜藻褐藻聚糖硫酸酯粗品分离纯化得到TF1、TF2、TF3、TF4 4个组分,其硫酸基团含量分别为9.62%、11.51%、14.51%和13.62%;单糖组成分析显示,粗多糖主要由岩藻糖(Fuc)、葡萄糖(Glu)、甘露糖(Man)和半乳糖(Gal)组成,各纯化组分的单糖组成含量差异较大;红外光谱分析显示,TF及其纯化组分在1 250 cm-1附近出现强吸收峰,为S=O(硫酸基)的吸收峰,证明TF及其纯化组分均含有硫酸基团,且TF、TF3、TF4的硫酸基主要连接于岩藻糖的C-2或C-3位置,TF1、TF2主要连接于岩藻糖的C-4位置;动物试验显示,灌胃不同剂量TF及其纯化组分,均能显著提高肝损伤小鼠肝脏或血清中SOD、GSH-Px和TNF-α的活性(P<0.05),显著降低MDA、ALT和AST指标的水平(P<0.05),肝组织切片显示,肝细胞病变程度较模型组明显减轻。研究表明,铜藻褐藻聚糖硫酸酯粗品经分离纯化得到4个组分,总体上具有显著的抗氧化和保肝护肝作用。

Abstract

In order to explore the structural characteristics and hepatoprotective activity of fucoidan from seaweed Sargassum horneri, four purified components TF1, TF2, TF3 and TF4 of the seaweed were obtained by DEAE-Sepharose Fast Flow anion exchange column.The monosaccharide composition and structural characteristics of crude polysaccharide(TF)and its purified components were analyzed, separated and purified by high performance liquid chromatography and infrared spectroscopy.An animal model of liver damage was established in mice with liver damage induced by CCl4 in the gavage test to evaluate the liver-protecting function of fucoidan from the seaweed.The results showed that there was 9.62% of TF1, 11.51% of TF2, 14.51% of TF3 and 13.62% of TF4 in the crude extraction of sulfate.The monosaccharide composition analysis revealed that the monosaccharides were primarily composed of fucose, glucose, mannose, and galactose, quite difference in the those of TF1-TF4.The IR spectrum indicated that all fucoidan fractions contained sulfate substitute with absorption peaks of S=O(1 250 cm-1).The further absorption peaks indicated that the sulfate substitutes of TF, TF3 and TF4 were mainly connected to the position of C-2 or C-3, while those of TF1 and TF2 were mainly connected to the position of C-4.The animal experiment results confirmed that TF, TF1-TF4 significantly inhibited the levels of malondialdehyde(MDA), and activities of aspartate aminotransferase(AST), and alanine aminotransferase(ALT), and increased the activities of superoxide dismutase(SOD), glutathione peroxide(GSH-Px)and tumor necrosis factor(TNF-α)in mice liver, proving that the fucoidan groups all had significant antioxidant effects.The mouse serum immune indicators indicated that all the level of TNF-α of all the fucoidan groups was significantly increased(P<0.05), and the increases in ALT and AST levels were significantly inhibited except the low-dose group of TF4(P<0.05).The findings indicated that fucoidan separated and purified from weaweed Sargassum horneri contained 4 components showing significant liverprotection effect.

关键词

铜藻 / 褐藻聚糖硫酸酯 / 分离纯化 / 结构组成 / 保肝作用

Key words

Sargassum horneri / fucoidan; purification / structural composition / hepatoprotective activity

引用本文

导出引用
卓丹琪, 张雪楠, 李雨晴, 刘诗萌, 任丹丹, 何云海, 汪秋宽. 铜藻褐藻聚糖硫酸酯的分离纯化、结构组成及保肝护肝作用[J]. 大连海洋大学学报, 2022, 37(2): 295-303 https://doi.org/10.16535/j.cnki.dlhyxb.2021-304
ZHUO Danqi, ZHANG Xuenan, LI Yuqing, LIU Shimeng, REN Dandan, HE Yunhai, WANG Qiukuan. Purification,structural characteristics and hepatoprotective effects of fucoidan from seaweed Sargassum horneri[J]. Journal of Dalian Fisheries University, 2022, 37(2): 295-303 https://doi.org/10.16535/j.cnki.dlhyxb.2021-304
中图分类号: S 917    TS 254   

基金

国家重点研发计划“蓝色粮仓科技创新”项目(2019YFD0900501);财政部和农业农村部:国家现代农业产业技术体系资助(CARS-46)

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