山羊β-乳球蛋白分离及免疫反应性比较研究  

Comparative study ofβ-lactoglobulin isolation and immunoreactivity in goats

作　　者：薛海燕[1] 张磊[1] 周妍 马军 贺宝元 孟毅 XUE Hai-yan;ZHANG Lei;ZHOU Yan;MA Jun;HE Bao-yuan;MENG Yi(School of Food Science and Engineering,Shaanxi University of Science&Technology,Xi′an 710021,China;College of Bioresources Chemical and Materials Engineering,Shaanxi University of Science&Technology,Xi′an 710021,China;Shaanxi Jinniu Dairy Co.,Ltd.,Weinan 714000,China)

机构地区：[1]陕西科技大学食品科学与工程学院,陕西西安710021 [2]陕西科技大学轻工科学与工程学院,陕西西安710021 [3]陕西金牛乳业有限公司,陕西渭南714000

出　　处：《陕西科技大学学报》2023年第5期57-63,共7页Journal of Shaanxi University of Science & Technology

基　　金：陕西省西安市科技计划-农业科技创新工程项目(20193035YF023NS023)。

摘　　要：β-乳球蛋白(β-lactoglobulin,β-Lg)是牛乳中最主要的过敏原,而羊乳β-Lg序列及结构与牛乳不同,其免疫反应性有待探讨.采用化学沉淀法与超滤离心法对羊乳β-Lg进行分离纯化,以牛羊乳β-Lg分别致敏利用Balb/c小鼠,分别构建牛羊乳β-Lg过敏动物模型,分离小鼠淋巴细胞,探讨体外β-Lg刺激后相关白细胞介素(interleukin,IL)水平变化,分析其免疫反应性.结果显示,化学沉淀法中当pH值为3.1,NaCl浓度为10%时,分离出的β-Lg纯度最高,经Image J灰度值分析,其纯度达到96.70%;超滤离心法中10 min时β-Lg获得量较少,在20 min和30 min时,得到含有β-Lg的混合物,纯度别为25.29%和26.06%;羊乳β-Lg致敏组超敏反应现象系统评分均低于牛乳β-Lg组;羊乳β-Lg致敏组分离后小鼠淋巴细胞中细胞因子IL-2、IL-4水平显著上升,IL-6、IL-12水平显著变化,但升高幅度小,羊乳β-Lg致敏组细胞因子水平总体上升幅度显著低于牛乳β-Lg致敏组.因此,羊乳β-Lg免疫反应性显著低于牛乳β-Lg,为羊乳致敏性低于牛乳提供理论基础.β-lactoglobulin(β-Lg)is the most important allergen in bovine milk,while the sequence and structure of sheep milkβ-Lg is different from that of bovine milk,and its immunoreactivity needs to be explored.The immunoreactivity of bovine and sheep milkβ-Lg allergic animal models was investigated by using chemical precipitation and ultrafiltration centrifugation to isolate and purifyβ-Lg from sheep milk.The results showed that the purity ofβ-Lg isolated by the chemical precipitation method was the highest when the pH value was 3.1 and the concentration of NaCl was 10%,and the purity reached 96.70%by Image J grey value analysis;in the ultrafiltration centrifugation method,the amount ofβ-Lg obtained was less at 10 min,and at 20 min and 30 min,a mixture containingβ-Lg was obtained with the purity of 25.29%and 26.06%,respectively.The levels of cytokines IL-2 and IL-4 in mouse lymphocytes increased significantly in the sheep milkβ-Lg sensitized group,and the levels of IL-6 and IL-12 changed significantly,but the increase was small,and the overall increase of cytokine levels in the sheep milkβ-Lg sensitized group was significantly lower than that in the cow milkβ-Lg sensitized group.The overall increase in cytokine levels in the goat milkβ-Lg sensitized group was significantly lower than that in the cow milkβ-Lg sensitized group.Therefore,the immunoreactivity of goat milkβ-Lg was significantly lower than that of cow milkβ-Lg,providing a theoretical basis for the lower sensitization of goat milk than cow milk.

关 键 词：乳清蛋白 细胞因子 BALB/C小鼠 动物模型 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程]