仙人掌多糖结构分析及抗凝血活性研究(英文)

Structural Analysis and Anticoagulant Activity of Polysaccharides from Opuntia milpa alta

[目的]分析仙人掌多糖组分和结构,并研究其抗凝血活性。[方法]以米邦塔仙人掌为试验材料,通过热水浸提、醇沉制得粗多糖CP,经交换层析和凝胶层析纯化,研究其单糖组成和重均分子质量分布与主链结构,并对多糖组分抗凝血活性进行了初步研究。[结果]对仙人掌粗多糖分离纯化后共得到3个多糖组分WSP1、WSP2a和WSP3,重均分子量分别为2.32×106、1.24×106和7.92×106。对氨基苯甲酸(PMP)衍生化高效液相色谱法检测表明,WSP1主要成分为半乳糖;WSP2a由鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖、葡萄糖醛酸和半乳糖醛酸组成,含量分别为6.3%、32.3%、12.9%、1.5%、4.8%、37.1%、0.64%和4.4%;WSP3主要成分为阿拉伯糖、半乳糖和木糖以及少量鼠李糖、甘露糖、糖醛酸。WSP2能延长活化部分凝血酶原时间(APTT)和凝血酶时间(TT),而对凝血酶原时间(PT)的影响不显著,说明其是通过影响内源性凝血系统而发挥抗凝血作用。纤维蛋白原转化纤维蛋白的抑制试验初步表明WSP2a和WSP3具有抗凝血活性。甲基化分析显示WSP2a含有1,4-连接的半乳糖酸,1,2-连接的鼠李糖和1,2,4-连接的鼠李糖构成的主链。[结论]该研究为仙人掌多糖资源优化和深度开发提供了试验依据。

[Objective] To analyze the components and structure of Opuntia milpa alta polysaccharides, and to study its anticoagulant activity. [Method] The crude polysaccharides （CP） were prepared from Opuntia milpa alta using hot water extraction and ethanol precipitation method, purified by anion-exchange chromatography and gel filtration chromatography to study its monosaccharide composition, weight-average molecular weight distribution and main chain structure. And a preliminary study was conducted on the anticoagulant activity of polysaccharide components. [Result] The polysaccharide components WSP1, WSP2a and WSP3 were obtained from CP by the anion-exchange chromatography and gel filtration chromatography. WSP1 was mainly composed of galactose with a weight-average molecular weight of 2.32×106. WSP2a consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid, the contents were 6.3%, 32.3%, 12.9%, 1.5%, 4.8%, 37.1%, 0.64% and 4.4%, respectively. WSP3 mainly composed of arabinose, xylose, rhamnose and trace of glucose, mannose, uronic acid. The weight-average molecular weight of WSP2a and WSP3 was 1.24×106 and 7.92×106, respectively. WSP2 prolonged the activated partial thromboplatin time （APTT） and thrombin time （TT）, while had no significant effect on the prothrombin time （PT） in vitro, indicating that anticoagulant activity of WSP2 functions by affecting the intrinsic coagulation system. The test of inhibition to the conversion of fibrinogen into thrombin showed that WSP2a and WSP3 had obvious anticoagulant activity. Methylation analysis indicated that the main chain of WSP2a was constituted of 1, 4-GalA, 1, 2-Rha and 1, 2, 4-Rha. [Conclusion] This study provided experimental basis for the optimization and development of polysaccharide resources from cactus