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干酪乳杆菌胞外多糖诱导小鼠骨髓来源树突细胞成熟以及分泌IL-6、TGF-β和IL-23 预览

Lactobacillus casei Exopolysaccharide Induces Maturation and IL-6,TGF-βand IL-23 Secretion of Mouse Bone Marrow-Derived Dendritic Cells
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摘要 为研究干酪乳杆菌(Lactobacillus casei)胞外多糖(exopolysaccharide,EPS)促进体外培养的小鼠未成熟骨髓来源树突细胞(bone marrow-derived dendritic cells,BMDCs)成熟的作用,分析不同剂量的EPS对BMDCs分泌细胞因子白细胞介素(interleukin,IL)-6、转化生长因子-β(transforming growth factor-β,TGF-β)和IL-23的影响,并检测EPS对BMDCs抗原提呈能力的影响。首先从干酪乳杆菌中分离纯化出EPS,并检测EPS的纯度;把从BALB/c小鼠体内分离出的骨髓细胞分化为BMDCs,未成熟BMDCs分别与磷酸盐缓冲液、不同剂量EPS和脂多糖(lipopolysaccharide,LPS)体外培养,采用流式细胞法检测了BMDCs成熟表面标记物MHC II和CD86的表达,酶联免疫吸附检测(enzyme-linked immunosorbent assay,ELISA)法分析了IL-6、TGF-β和IL-23的表达量,CCK-8法检测了BMDCs与小鼠脾淋巴细胞的混合培养体系中淋巴细胞的增殖率。结果表明:分离出EPS的纯度约为95%;EPS能够上调BMDCs表面MHC II和CD86的表达量;EPS可以显著促进BMDCs分泌IL-6、TGF-β和IL-23(P<0.05),但促进水平低于LPS;EPS可以明显促进BMDCs刺激异基因淋巴细胞的增殖。综上所述,在体外实验中,干酪乳杆菌EPS能够促进BALB/c小鼠BMDCs的成熟,诱导BMDCs分泌与辅助性T17细胞分化和增殖相关的细胞因子IL-6、TGF-β和IL-23,并且可以增强BMDCs的抗原提呈能力。 This study was undertaken in order to understand the effect of Lactobacillus casei exopolysaccharide(EPS)on the maturation of mouse bone marrow-derived dendritic cells(BMDCs)in vitro,the effect of different doses of EPS on interleukin(IL)-6,transforming growth factor-β(TGF-β)and IL-23 secretion in BMDCs,and the effect of EPS on the antigen-presenting function of BMDCs.L.casei EPS was separated and purified from the fermentation broth,and its purity was determined.The bone marrow cells isolated from BALB/c mice were differentiated into BMDCs,and immature BMDCs were treated with phosphate buffer saline(PBS),EPS and lipopolysaccharide(LPS),respectively.The phenotypic maturation of BMDCs,MHC II and CD86 were analyzed using a flow cytometer,the production of cytokines was analyzed by enzyme-linked immunosorbent assay(ELISA),and the proliferation rate of splenic lymphocytes was analyzed by cell counting kit-8(CCK-8)assay in a mixed culture system with BMDCs and mouse splenic lymphocytes.The results showed that the purity of EPS was 95%.The EPS up-regulated the expression of MHC II and CD86 on the surface of BMDCs.Meanwhile,the EPS significantly promoted the production of IL-6,TGF-βand IL-23(P<0.05)but not so effectively as LPS.The EPS also obviously promoted BMDCs-stimulated proliferation of allogenic lymphocytes.In conclusion,L.casei EPS can promote the maturation of BMDCs in vitro,induce BMDCs to secrete cytokines associated with the differentiation and proliferation of Th17 cells,and enhance the antigen-presenting function of BMDCs.
作者 任琦琦 任皓威 杨翠翠 刘宁 REN Qiqi;REN Haowei;YANG Cuicui;LIU Ning(Key Laboratory of Dairy Science,Ministry of Education,College of Food Science,Northeast Agricultural University,Harbin 150030,China)
出处 《食品科学》 EI CAS CSCD 北大核心 2019年第5期175-182,共8页 Food Science
基金 教育部创新团队基金项目(IRT-0959-202) 哈尔滨市科技创新人才研究专项资金项目(2011RFXXN042).
关键词 干酪乳杆菌 胞外多糖 树突细胞 成熟 细胞因子 分泌 抗原提呈 Lactobacillus casei exopolysaccharide dendritic cells maturation cytokines secretion antigen presentation
作者简介 第一作者:任琦琦(1993—)(ORCID: 0000-0002-6108-7838),女,硕士研究生,研究方向为食品营养及加工。E-mail: rqqrenqiqi@163.com;通信作者:刘宁(1960-)(ORCID: 0000-0003-0337-5409),男,教授,博士,研究方向为食品营养及加工。E-mail: ningliuneau@outlook.com.
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