纳溪特早茶‘乌牛早’茶多糖提取纯化及免疫活性研究  

Extraction,Purification and Immune Activity of Polysaccharide from Naxi Wuniu Tea

作　　者：赖玲 杨潇 尹琳[1] 陈小雨 阳辉蓉 彭取敏 刘军 曾英杰 LAI Ling;YANG Xiao;YIN Lin;CHEN Xiaoyu;YANG Huirong;PENG Qumin;LIU Jun;ZENG Yingjie(College of Food Science and Technology,Xinan Minzu Univercity,Chengdu 610041,China;Dadukou Town,Naxi District,Luzhou 646329,China;Sichuan Dalit Morning Tea Co.,Ltd.,Luzhou 646318,China)

机构地区：[1]西南民族大学食品科学与技术学院,四川成都610041 [2]泸州市纳溪区大渡口镇,四川泸州646329 [3]四川大里特早茶有限公司,四川泸州646318

出　　处：《食品工业科技》2025年第6期231-241,共11页Science and Technology of Food Industry

基　　金：西南民族大学研究生创新型科研项目(ZD2023050);四川省自然科学基金项目(2023NSFSC1200);四川省天府峨眉计划青年科技人才项目(川峨眉第2260号);攀西特色作物研究与利用四川省重点实验室资助课题(ZYN2023037)。

摘　　要：目的:本研究旨在对纳溪乌牛特早茶多糖进行分离纯化,并进一步研究其抗氧化和免疫活性。方法:首先,采用热水浸提法获取乌牛茶多糖,采用DEAE-52和葡聚糖凝胶G200进一步纯化乌牛茶多糖。然后,对所获取多糖的基本理化性质进行研究;并通过检测乌牛茶多糖的1,1-二苯基-2-三硝基苯肼(2,2-Diphenyl-1-picrylhydrazyl,DPPH)自由基清除能力、2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS)阳离子自由基清除能力、氧化自由基吸收能力(oxygen radical absorbance capacity,ORAC)评价其抗氧化活性。接着,对乌牛茶多糖的安全性进行分析检测,具体包括:内毒素检测,小鼠巨噬细胞RAW264.7细胞和人胚胎肾细胞293(人胚胎肾细胞293,HEK293)毒性检测。最后,评价乌牛茶多糖对RAW264.7细胞因子(白细胞介素6,Interleukin-6,IL6;肿瘤坏死因子α,tumor necrosis factorα,TNF-α;NO)释放及对应mRNA分泌水平的影响,进而获取乌牛茶多糖免疫增强活性。结果:采用DEAE-52阴离子交换树脂和葡聚糖凝胶Sephadex G-200从纳溪乌牛早茶中分离得到纯化后乌牛茶多糖(WNp);理化分析表明WNp纯度为90.26%,蛋白质含量为1.37%,且不具有三螺旋结构;WNp由摩尔比为0.4:32.6:2.3:4.7:8.7:0.3:40.3的葡萄糖、半乳糖、阿拉伯糖、甘露糖、鼠李糖、葡萄糖醛酸和半乳糖醛酸组成的分子量为82.61 kDa的多糖;WNp对DPPH和ABTS+自由基具有显著的清除能力并具有较高的ORAC值,即具有较好的抗氧化性;最后,WNp可上调控细胞因子(IL-6、TNF-α)分泌和NO释放的mRNA的表达量,从而促进细胞因子(IL-6、TNF-α)分泌和NO释放,进而激活RAW264.7细胞的初始免疫应答反应,达到增强免疫的效果。结论:研究结果为纳溪乌牛茶源的精深加工利用提供科学依据。Objective:The aim of this study was to isolate and purify the polysaccharide from Naxi Wuniu special morning tea,and the antioxidant and immune activities were further investigated.Methods:Firstly,the tea polysaccharides were extracted by hot water extraction method,and further purified by DEAE-52 and dextranan gel G200.Then,the basic physicochemical properties of polysaccharide were studied.And the antioxidant activities were evaluated by determined the 1,1 Diphenyl-2-trinitrobenzene hydrazine(DPPH)free radical scavenging capacity,2,2'-diazo-di-3-ethylbenzothiazolin-6-sulfonic acid(ABTS)cationic free radical scavenging capacity,and oxygen radical absorbance capacity(ORAC).Then,the safeties of tea polysaccharide were analyzed,including endotoxin detection,RAW264.7 cells and human embryonic kidney cell 293(HEK293)toxicity detection.Finally,the effects of tea polysaccharide on RAW264.7 cytokines(interleukin-6,IL6,tumor necrosis factor α,TNF-α,NO)release and the corresponding mRNA secretion levels,in order to obtain the immuneenhancing activity of tea polysaccharide.Results:A purified polysaccharide fraction WNp was obtained from Naxi Wuniu morning tea by using DEAE-52 anion exchange resin and Sephadex G-200.Physicochemical analysis showed that the purity of WNp was 90.26%,the protein content of WNp was 1.37%,and WNp did not contain triple helix structure.WNp was composed of glucose(Glc),galactose(Gal),arabinose(Ara),mannose(Man),rhamnose(Rha),glucuronic acid(GlcA)and galacturonic acid(GalA)at the mole ratio of 0.4:32.6:2.3:4.7:8.7:0.3:40.3,respectively.The molecular weight of WNp was 82.61 kDa.WNp showed significant scavenging abilities on DPPH and ABTS+free radicals and a high ORAC value,that was,it had good antioxidant activities.Finally,WNp could up-regulate the mRNA expression of cytokines(IL-6,TNF-α)and NO,thereby promoting cytokine secretion(IL-6,TNF-α)and NO release,and then activate the initial immune response of RAW264.7 cells,which achieved the immuno-enhancing activity.Conclusion:The research results

关 键 词：纳西乌牛特早茶 多糖 结构 抗氧化性 免疫活性 

分 类 号：TS209[轻工技术与工程—食品科学]