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女贞花水溶性多糖的分离、纯化、结构分析及凝血活性

Isolation, purification, structural analysis and coagulatory activity of water-soluble polysaccharides from Ligustrum lucidum Ait flowers.

作者信息

Yin Zhenhua, Zhang Wei, Zhang Juanjuan, Kang Wenyi

机构信息

Huanghe Science and Technology College, Zhengzhou, 450063, China.

Zhengzhou City Key Laboratory of Medicinal Resources Research, Zhengzhou, 450063, China.

出版信息

Chem Cent J. 2017 Oct 4;11(1):98. doi: 10.1186/s13065-017-0332-y.

Abstract

In this study, Ligustrum lucidum flowers as raw material, the extraction, isolation and coagulatory activity of polysaccharides were carried out for the first time. The crude polysaccharide was obtained by hot water extraction and ethanol precipitation, and preliminarily purified by Sevage method and D101 macroporous resin. Then the polysaccharide was further purified by DEAE-52 cellulose and Sephadex G-100 column chromatography, respectively. The structural characteristics were detected by LC, GC, FT-IR and NMR. Furthermore, the coagulatory activity of the polysaccharides were investigated by APTT, TT, PT and FIB assays in vitro. The results demonstrated that four polysaccharides were isolated from flowers of L. lucidum, named as LLP-1a, LLP-1b, LLP-2 and LLP-3, and the yields were 0.039, 0.0054, 0.0055 and 0.017%, respectively based on the weight of the dried flowers. The four polysaccharides components were free of nucleic acids and proteins, and their average molecular weights were 25,912, 64,919, 3,940,246 and 2,975,091 g/mol, respectively. The monosaccharide compositions of LLp-1a were L-rhamnose, L-arabinose, D-xylose, D-glucose and D-galactose (molar ratio of 3.16: 2.46: 1.00: 7.27: 4.22). Only D-galactose was detected from LLp-1b. LLp-2 was composed of L-arabinose, D-glucose and D-galactose (molar ratio of 1.28:1.32:1.00). LLp-3 was composed of L-rhamnose, L-arabinose, D-xylose, D-glucose and D-galactose (molar ratio of 5.85: 2.21: 2.23: 1.00: 2.25). Coagulation assays indicated that LLp-1a and LLp-3 had good anticoagulant effect in vitro, while LLp-1b showed procoagulant activity.

摘要

本研究首次以女贞花为原料,开展了多糖的提取、分离及凝血活性研究。通过热水提取和乙醇沉淀获得粗多糖,并依次用Sevage法和D101大孔树脂进行初步纯化。然后分别用DEAE - 52纤维素柱色谱和Sephadex G - 100柱色谱对多糖进一步纯化。采用液相色谱(LC)、气相色谱(GC)、傅里叶变换红外光谱(FT - IR)和核磁共振(NMR)对其结构特征进行检测。此外,通过活化部分凝血活酶时间(APTT)、凝血酶时间(TT)、血浆凝血酶原时间(PT)和纤维蛋白原(FIB)体外检测法对多糖的凝血活性进行研究。结果表明,从女贞花中分离得到4种多糖,分别命名为LLP - 1a、LLP - 1b、LLP - 2和LLP - 3,基于干花重量计算,其得率分别为0.039%、0.0054%、0.0055%和0.017%。这4种多糖组分不含核酸和蛋白质,其平均分子量分别为25,912、64,919、3,940,246和2,975,091 g/mol。LLp - 1a的单糖组成为L - 鼠李糖、L - 阿拉伯糖、D - 木糖、D - 葡萄糖和D - 半乳糖(摩尔比为3.16 : 2.46 : 1.00 : 7.27 : 4.22)。LLp - 1b仅检测到D - 半乳糖。LLp - 2由L - 阿拉伯糖、D - 葡萄糖和D - 半乳糖组成(摩尔比为1.28 : 1.32 : 1.00)。LLp - 3由L - 鼠李糖、L - 阿拉伯糖、D - 木糖、D - 葡萄糖和D - 半乳糖组成(摩尔比为5.85 : 2.21 : 2.23 : 1.00 : 2.25)。凝血试验表明,LLp - 1a和LLp - 3在体外具有良好的抗凝作用,而LLp - 1b表现出促凝血活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6db/5628082/681cd3cdb08b/13065_2017_332_Fig1_HTML.jpg

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