二十二碳六烯酸对IgE介导的过敏反应的调节-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Regulation of Docosahexaenoic Acid Against IgE-Mediated Allergic Reaction

作者:: 徐乐1; 2; 3; 吴梦娜1; 2; 3; 邵鹏1; 2; 3; 张丽媛1; 2; 3; 黄世英1; 2; 3; 林静瑜1; 2; 3; 李桂玲1; 2; 3; (1.集美大学海洋食品与生物工程学院，福建厦门 361021；2.福建省海洋功能食品工程技术研究中心，福建厦门 361021；3.厦门市海洋功能食品重点实验室，福建厦门 361021)

Author(s):: XU Le1; 2; 3; WU Mengna1; 2; 3; SHAO Peng1; 2; 3; ZHANG Liyuan1; 2; 3; HUANG Shiying1; 2; 3; LIN Jingyu1; 2; 3; LI Guiling1; 2; 3; (1.College of Ocean Food and Biological Engineering,Jimei University,Xiamen 361021,China;2.Marine Functional Food Engineering Technology Research Center of Fujian Province,Xiamen 361021,China;3.Xiamen Key Laboratory of Marine Functional Food,Xiamen 361021,China)

Keywords:: docosahexaenoic acid; allergic response; cytokines; unsaturated fatty acid; signal pathway

摘要:: 为了探究二十二碳六烯酸（docosahexaenoic acid，DHA）对过敏反应的调节作用和可能的机制，以IgE/BSA诱导的RBL-2H3细胞为研究对象，通过测定β-氨基己糖苷酶的释放，建立细胞过敏模型；进一步用ELISA法测定组胺及细胞因子释放量，并用Fluo-3 AM荧光探针法测定钙离子浓度；用定量逆转录聚合酶链式反应（quantitative reverse transcription polymerase chain reaction，qRT-PCR）测定细胞因子及高亲和力受体FcεRI亚基的表达水平；用蛋白质免疫印迹法测定FcεRI信号通路蛋白的表达。结果显示，RBL-2H3敏化细胞经DHA处理后，其β-氨基己糖苷酶、组胺的释放率及细胞内钙离子浓度显著降低，说明DHA具有抗过敏作用。qRT-PCR检测结果表明，DHA处理能显著降低FcεRIα亚基的表达，但对FcεRIβ和FcεRIγ基因表达无显著影响。进一步的Western Blot检测结果显示，DHA能显著抑制RBL-2H3敏化细胞中PLCγ/JNK和Akt/NF-κB信号通路的活性，从而降低下游炎症细胞因子I-4和TNF-α的释放量。可见，DHA通过抑制PLCγ/JNK和Akt/NF-κB信号通路的活性降低IgE介导的RBL-2H3细胞过敏反应。

Abstract:: To explore the regulatory effect of DHA on allergic reaction and the possible mechanism,IgE/BAS-induced RBL-2H3 cells were used as the study object and the release of β-hexosidase was measured to establish the allergy cellular model.Furthermore,histamine and cytokines were determined by ELISA.Calcium ion concentration was determined using the Fluo-3 AM fluorescence probe.The expression levels of cytokines and high affinity receptor FcεRI subunits were determined by quantitative reverse transcription polymerase chain reaction.The expression of FcεRI signal pathway protein was determined by protein immunoblotting method.The results showed that the release amount of β-hexosidase and histamine,as well as the intracellular calcium concentration of sensitized RBL-2H3 cells were significantly reduced upon DHA treatment,suggesting that DHA had an antiallergic effect.Moreover,the qRT-PCR results showed that DHA treatment significantly reduced the gene expression of FcεRIα subunits without obvious effect on FcεRIβ and FcεRIγ subunit expression.Further Western blot studies showed that DHA significantly inhibited the activity of the PLCγ/JNK and Akt/NF-κB signal pathways in sensitized RBL-2H3 cells,thereby reducing the release of downstream inflammatory cytokines IL-4 and TNF-α.In summary,DHA inhibits IgE-mediated allergic reactions in RBL-2H3 cells by inhibiting the activity of PLCγ/JNK and Akt/NF-κB signal pathways.