板栗果酒多糖的提取纯化及其抗疲劳活性

Extraction and Purification of Polysaccharide from Chestnut Fruit Wine and Its Anti-fatigue Activity

目的:本研究对板栗果酒多糖进行了提取纯化和结构表征,并对其抗疲劳活性进行了研究。方法:采用超声波辅助水提醇沉法提取板栗果酒粗多糖,用超滤技术进行纯化分离。利用离子色谱、凝胶渗透色谱、紫外光谱和红外光谱等方法对其结构进行分析,并通过小鼠负重游泳实验来研究其抗疲劳的功效。结果:经超滤膜分离纯化得到板栗果酒多糖CFWP-1(>100 kU)和CFWP-2(<100 kU)。这两种多糖都属于非均一多糖,CFWP-1的重均分子量为132351 Da和664 Da,CFWP-2的重均分子量达133619 Da和664 Da。单糖分析结果表明,CFWP-1和CFWP-2主要由阿拉伯糖、半乳糖、葡萄糖、半乳糖醛酸、甘露糖醛酸5种单糖成分组成,且这两种多糖中葡萄糖含量均达98%以上,属于中性葡聚糖。紫外光谱表明,2种多糖的纯度较高。红外光谱分析显示,这两种多糖都有多糖典型的吸收峰。抗疲劳试验表明,CFWP-1和CFWP-2均能显著提高小鼠血清中的超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)(P<0.05)的活力,并能明显地降低乳酸脱氢酶(Lactate dehydrogenase,LDH)、血清尿素氮(Serum urea nitrogen,BUN)、乳酸(Bloodlactate, BLA)、丙二醛(malondialdehyde, MDA)和肌酸激酶(creatinekinase, CK)的水平(P<0.05),可以显著提升糖原储备水平,对小鼠骨骼肌具有良好的保护作用,且CFWP-2的效果优于CFWP-1,并呈剂量依赖性。结论:从板栗果酒中提取出的多糖具有较好的抗疲劳作用。

Objective:In this study,the polysaccharide of chestnut fruit wine was extracted,purified and characterized,and its anti-fatigue activity was studied.Methods:The crude polysaccharide of chestnut fruit wine was extracted by ultrasonic-assisted water extraction and alcohol precipitation,and purified and separated by ultrafiltration technology.Its structure was analyzed by ion chromatography,gel permeation chromatography,ultraviolet spectroscopy and infrared spectroscopy,and its anti-fatigue effect was studied by weight-bearing swimming test in mice.Results:CFWP-1(>100 kU)and CFWP-2(<100 kU)were isolated and purified by ultrafiltration membrane.Both of these polysaccharides were heterogeneous,with the average molecular weight of CFWP-1 being 132351 Da and 664 Da,and the average molecular weight of CFWP-2 being 133619 Da and 664 Da.The results of monosaccharide analysis showed that CFWP-1 and CFWP-2 were mainly composed of arabinose,galactose,glucose,galacturonic acid and mannouronic acid,and the glucose content in these two polysaccharides was more than 98%,which belonged to neutral glucan.UV spectra showed that the purity of the two polysaccharides was high.Infrared spectroscopy analysis showed that both polysaccharides had typical absorption peaks of polysaccharides.Anti-fatigue experiments showed that both CFWP-1 and CFWP-2 could significantly increase the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in serum of mice(P<0.05).The levels of lactate dehydrogenase(LDH),serum urea nitrogen(BUN),blood lactate(BLA),malondialdehyde(MDA)and creatine kinase(CK)were significantly decreased(P<0.05),and the glycogen storage level was significantly increased.It had a good protective effect on skeletal muscle in mice.The effect of CFWP-2 was better than that of CFWP-1 in a dose-dependent manner.Conclusion:The polysaccharide extracted from chestnut fruit wine would had good anti-fatigue effect.