双歧杆菌对TregTh17细胞平衡及中国对虾蛋白致敏性的影响
目的 通过体内实验研究婴儿双歧杆菌(Bifidobacterium infantis 13.085)和雷帕霉素对中国对虾原肌球蛋白致敏BALB/c小鼠过敏反应的治疗作用, 从Treg/Th17细胞平衡及相关细胞因子角度探讨其缓解过敏的免疫调节作用机制。方法 将中国对虾原肌球蛋白和弗氏佐剂混合液腹腔注射诱发BALB/c小鼠致敏, 建立动物过敏模型。将实验小鼠随机分为正常对照组、致敏对照组、双歧杆菌治疗组、雷帕霉素治疗组。观察分析小鼠过敏症状, 采用ELISA测定小鼠血清中特异性IgE、IgG2a的含量, 采用流式细胞术测定脾脏T淋巴细胞亚群(Treg、Th17)数量, 采用荧光定量PCR测定脾脏中Treg型和Th17型细胞因子和转录因子的表达量。结果 第56天实验周期结束后结果发现, 相比于致敏对照组, 双歧杆菌和雷帕霉素治疗组小鼠过敏症状有明显的缓解, 血清中特异性IgE显著降低(P<0.05), 脾脏Treg/Th17比值显著升高(P<0.05), Th17型细胞因子IL-17A mRNA表达水平显著降低, Treg型细胞因子Foxp3 mRNA表达水平显著升高。此外, 不同剂量的雷帕霉素治疗组缓解过敏反应存在剂量差异性。结论 双歧杆菌13.085和雷帕霉素能有效缓解小鼠过敏症状, 其作用可能通过平衡Treg/Th17细胞亚群数量, 促进Treg型细胞因子表达而抑制Th17型细胞因子分泌有关。
Objective To study the therapeutic effect of Bifidobacterium infantis 13.085 and rapamycin on suppression of shrimp tropomyosin (TM) sensitization in BALB/c mice, and to explore the mechanism of immunoregulation of hypersensitivity from the perspective of the balance of Treg/Th17 ratio and expression of Treg- and Th17-associated cytokines. Methods Five groups (Positive, B. infantis, and different doses of rapamycin) of BALB/c mice were immunized by intraperitoneal (IP) injection of purified TM together with incomplete freund's adjuvant. Negative control animals received equal amounts of sterile PBS with freund's adjuvant at each sensitization and challenge point. Allergic symptoms were evaluated. The levels of specific IgE and IgG2a in serum were determined by ELISA. The amount of Treg and Th17 cell subsets were analyzed by flow cytometry. The mRNA expression levels of Treg- and Th17-associated cytokines and transcriptional factors were detected by quantitative PCR. Results At the end of experimental period (d 56), the allergic symptoms of the mice treated with B. infantis 13.085 and rapamycin were obviously relieved, the levels of specific IgE (P<0.05) as well as IL-17A mRNA expression were significantly reduced, while the Treg/Th17 ratio was remarkably increased in B. infantis treatment groups compared to positive control. The level of Foxp3 mRNA expression was significantly enhanced in B. infantis therapy group compared to positive control. In addition, different doses of rapamycin treatment group had different dose response in relieving allergic reactions. Conclusion Therapeutic administration of B. infantis 13.085 and rapamycin can effectively alleviate TM-stimulated allergic inflammation and symptoms in mice, which may attribute to the balance of Treg/Th17 ratio and increase the expression of Treg-associated cytokines Thereby inhibiting the secretion of Th17 type cytokines.
标题：双歧杆菌对TregTh17细胞平衡及中国对虾蛋白致敏性的影响
英文标题：Effect of Bifidobacterium infantis on Treg/Th17 balance and Fenneropenaeus chinensis tropomyosin-stimulated sensitization in mice
作者：
彭吉祥 浙江工商大学食品与生物工程学院
王翀 浙江工商大学食品与生物工程学院
王彦波 浙江工商大学食品与生物工程学院
傅玲琳 浙江工商大学食品与生物工程学院
中文关键词:中国对虾原肌球蛋白,双歧杆菌,雷帕霉素,过敏,Treg/Th17比例,
英文关键词:Fenneropenaeus chinensis tropomyosin,Bifidobacterium infantis,rapamycin,sensitization,Treg/Th17 ratio,
发表日期：2016-12-27
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