淡水鲈鱼小清蛋白致敏性及其对小鼠肠道菌群的影响
目的 研究淡水鲈鱼小清蛋白(parvalbumin, PV)的致敏性, 同时探索其对小鼠肠道菌群的影响。方法36只C3H/HeJ小鼠按体重随机分为3组, 分别为空白对照组(CK组)、卵清白蛋白(ovalbumin, OVA)致敏对照组(OVA组)和小清蛋白致敏组(PV组)。使用生理盐水、OVA溶液和PV溶液通过经口灌胃在6周时间内进行致敏, 实验最后一天进行大剂量刺激, 测定小鼠的体温变化、血清抗体水平和相关细胞因子水平, 取腹腔灌洗液测定其中蛋白的含量以评价小鼠血管渗透性的变化, 利用流式细胞术测定各组小鼠脾脏(SP)和肠系膜淋巴结(MLN)中T细胞亚群的水平。取小鼠粪便, 对肠道菌群进行16S rRNA高通量测序, 探索小清蛋白对小鼠肠道菌群的影响。结果 与空白对照组相比, OVA小鼠血清抗体、细胞因子和免疫组织Th2相比水平均显著升高(P<0.05), 且表现出明显过敏症状。PV处理也使小鼠表现出明显的食物过敏症状, 但仅显著升高了小鼠血清免疫球蛋白G1(Immunoglobulin G1, IgG1)和白细胞介素10(Interleukin 10, IL-10)的水平(P<0.05)。16S rRNA高通量测序结果显示, OVA处理升高了小鼠肠道中属于拟杆菌门和变形菌门群落的水平, 而PV处理升高了属于厚壁菌门菌群的丰度。结论 淡水鲈鱼小清蛋白对小鼠具有一定的致敏性, 但其致敏机理及对肠道菌群的影响作用与OVA不同。
Objective To explore the allergenicity of parvalbumin (PV) derived from freshwater bass and its effect on the intestinal flora of mice. Methods 36 female C3H/HeJ mice were divided into 3 groups randomly as control group (CK), ovalbumin group (OVA) and PV group (PV) respectively. Saline, OVA and PV were introduced to mice byoral gavage for 4 times in 6 weeks. At the end of the experiment, body temperature of mice was detected, sera were collected for measurement of antibody and related cytokines, spleen (SP) and mesenteric lymph nodes (MLN) werecollectedto determine levels of Th1, Th2 and Treg cell subsets through flow cytometry. Fecal of every mouse was collected for 16S rRNA sequencing to explore the effect of PV on mice intestinal microbiota. Results Compared with the CK, serum levels of antibodies, cytokines and Th2 subsets in SP and MLN of OVA treated mice were significantly increased (P<0.05), and allergic symptoms were observed in these mice. PV treated mice also showed significant food allergy symptoms, but only significantly increased serum Immunoglobulin G1 (IgG1) and Interleukin 10 (IL-10) (P<0.05). Results of 16S rRNA sequencing showed that OVA treatment increased the level of families belonging to Bacteroides and Proteobacteria in the intestine of mice, while PV treatment increased the abundance of bacteria belonging to Firmicutes. Conclusion Oral administration of PV extract from freshwater bass cause allergy in mice, and its allergic mechanism and influences on intestinal microbiota of mice are different from OVA.
标题：淡水鲈鱼小清蛋白致敏性及其对小鼠肠道菌群的影响
英文标题：Allergenicity and influnce on mice intesitinal microbiota of parvalbumin from freshwaterbass
作者：
卢睿祺 首都医科大学公共卫生学院
周催 首都医科大学公共卫生学院
中文关键词:淡水鲈鱼,小清蛋白,经口致敏,肠道菌群,
英文关键词:freshwater bass,parvalbumin,oral sensitization,intestinal microbiota,
发表日期：2020-05-11
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