水解乳清蛋白改善牛乳蛋白过敏的功效及机制  

Efficacy and Mechanism of Hydrolyzed Whey Protein in Ameliorating Cow's Milk Protein Allergy

作　　者：储小军 李归浦 熊丽娜 姜艳喜 郑华艳 刘东红[1] Chu Xiaojun;Li Guipu;Xiong Lina;Jiang Yanxi;Zheng Huayan;Liu Donghong(College of Biosystems Engineering and Food Science,Zhejiang University,Hangzhou 310058;Beingmate(Hangzhou)Food Research Institute Co.,Ltd.,Hangzhou 310053)

机构地区：[1]浙江大学生物系统工程与食品科学学院,杭州310058 [2]贝因美(杭州)食品研究院有限公司,杭州310053

出　　处：《中国食品学报》2024年第3期46-55,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：杭州市农业和社会发展重点项目(202203A11);国家重点研发计划项目(2019YFE0103900)。

摘　　要：目的:研究部分水解乳清蛋白(pHWP)联合不同剂量低聚果糖(FOS)和动物双歧杆菌Bb-12(Bb-12)对小鼠牛乳蛋白过敏(CMPA)的改善效果及免疫耐受调节机制。方法:4周龄雌性BALB/c小鼠随机被分成正常对照组、致敏对照组、p HWP和pHWP+FOS+Bb-12干预组。试验期间,干预组小鼠每天用含pHWP(15 mg/mL)或pHWP(15 mg/mL)+FOS(0.4/4.0mg/mL)+Bb-12(1×10^(5)/10^(6)CFU/mL)的水溶液替代常规饮用水进行饲喂干预;试验第7,14,21,28,35天,致敏对照组和干预组小鼠采用CMP(20 mg/只)+霍乱毒素(10μg/只)灌胃致敏,正常对照组小鼠灌胃无菌生理盐水;试验第42天,对所有小鼠进行CMP(50 mg/只)灌胃激发,并于激发后1 h内完成过敏症状评分,同时检测血清抗体和组胺含量、脾细胞增殖反应及上清液Th1、Th2和Treg相关细胞因子(INF-γ、IL-4和TGF-β)水平。结果:pHWP+FOS+Bb-12干预组小鼠过敏症状评分(0~1)、血清总IgE(169.4~272.7 ng/mL)、CMP特异性IgE(sIgE)(0.038~0.414 OD)和sIgG1(0.252~0.379 OD)、组胺(19.56~27.35 ng/mL)、脾细胞增殖刺激指数(1.48~1.58)及上清液IL-4(18.48~29.60 pg/mL)均显著低于致敏对照组(P<0.05),而TGF-β(143.1~177.2 pg/mL)、INF-γ(189.5~230.2 pg/mL)和INF-γ/IL-4(6.46~12.71)均显著高于致敏对照组(P<0.05)。相较于p HWP,pHWP+FOS+Bb-12对CMPA的缓解效果更强,且以使用高剂量Bb-12(1×10^(6)CFU/mL)时最为显著,部分指标包括血清总IgE、CMP-sIgE、脾细胞上清液IL-4和INF-γ/IL-4与正常对照组均无显著差异。结论:pHWP联合FOS和Bb-12可通过提高CMP免疫耐受有效改善小鼠CMPA症状,其作用可能与阻断IgE、CMP-sIgE、sIgG1和组胺分泌,抑制脾细胞增殖及上调TGF-β和INF-γ/IL-4相关。Objective:To investigate the efficacy and immune tolerance regulation mechanism of partially hydrolyzed whey protein(pHWP)combined with different doses of fructooligosaccharides(FOS)and Bifidobacterium animalis subsp.lactis Bb-12(Bb-12)in alleviating cow's milk protein allergy(CMPA)in mice.Methods:The four-week-old female BALB/c mice were randomly assigned into normal control,sensitized control,pHWP and pHWP+FOS+Bb-12 intervention groups.During the experimental period(day 0-42),the mice in the intervention groups were continuously fed with an aqueous solution containing pHWP(15 mg/mL)or pHWP(15 mg/mL),FOS(0.4 or 4.0 mg/mL)and Bb-12(1×10^(5)or 10^(6) CFU/mL)instead of drinking water.Mice in the sensitized control group and intervention groups were sensitized by intragastric administration of sterile saline containing 20 mg CMP and 10μg cholera toxin on day 7,14,21,28 and 35,while mice in normal control group received only sterile saline.On day 42,all mice were oral challenged with 50 mg CMP,and allergic symptom scoring were conducted within 1 h after challenge.Serum antibody and histamine contents,as well as splenocyte proliferation and supernatant Th1,Th2 and Tregrelated cytokine(INF-γ,IL-4 and TGF-β)levels were assessed.Results:The allergic symptom scores(0-1),serum total IgE(169.4-272.7 ng/mL),CMP-specific IgE(sIgE)(0.038-0.414 OD),sIgG1(0.252-0.379 OD),and histamine contents(19.56-27.35 ng/mL),splenocyte proliferation stimulation indexes(1.48-1.58)and supernatant IL-4 levels(18.48-29.60 pg/mL)of mice in the pHWP+FOS+Bb-12 intervention groups were significantly lower than those in the sensitized control group(P<0.05),while the splenocyte supernatant TGF-β(143.1-177.2 pg/mL),INF-γlevels(189.5-230.2 pg/mL)and INF-γ/IL-4 ratios(6.46-12.71)were significantly higher than those in the sensitized control group(P<0.05).Compared with pHWP,pHWP+FOS+Bb-12 was more effective in alleviating CMPA,especially when high dose of Bb-12(1×10^(6) CFU/mL)was applied.No significant difference was found in serum total IgE,CMP-

关 键 词：部分水解乳清蛋白 低聚果糖 动物双歧杆菌Bb-12 牛乳蛋白过敏 免疫耐受 免疫调节 

分 类 号：TS201.21[轻工技术与工程—食品科学]