[关键词]
[摘要]
本文以坚果过敏蛋白为抗原,构建小鼠动物致敏免疫模型,探讨坚果过敏原的致敏机理,为坚果类食物安全评价和安全开发提供有效的实验依据。选取六种坚果为原料,以昆明小鼠为受试小鼠,坚果蛋白液以0.01 mg/g进行灌胃致敏,第33 d和34 d以0.02 mg/g激发致敏,通过测定小鼠血清IgE、IgG、腹腔肥大细胞及类胰蛋白酶和血象指标,研究其病理特征。结果表明,致敏激发后小鼠IgE与IgG最高水平为0.91和0.93,组胺释放率阳性组为58%,中剂量组为40%,高剂量组为55%;小鼠腹腔肥大细胞体外激发后脱颗粒高剂量组最多以及类胰蛋白酶活性也最高;中和高剂量组、阳性组白细胞数较高(9.7±6.1、9.6±2.4、9.2±2)×109/L,低剂量组、中剂量组和高剂量组的血小板数依次降低。说明致敏后小鼠凝血功能显著减弱,免疫系统受到侵袭,体内诱发产生IgE抗体,并启动过敏介质的释放机制,导致过敏反应Ⅰ型变态反应的发生。
[Key word]
[Abstract]
Nut allergenic protein was used as the antigen here to construct a mouse model of sensitization. The mechanism of sensitization of nut allergen was explored to provide an effective experimental basis for the safe development of nut foods and the evaluation of their safety. Six kinds of nuts were used as raw material and Kunming mice were used as test animals, which were fed with 0.01 mg/g nut protein extract by gavage to induce sensitization. On days 33 and 34, the mice were stimulated with 0.02 mg/g nut protein extract to induce sensitization, and pathological features were studied by measuring the serum IgE and IgG levels, the number of peritoneal mast cells, tryptase level, and hematological parameters in the mice. The results showed that the highest levels of IgE and IgG after sensitization in mice were 0.91 and 0.93, respectively, and histamine release rates in the positive, medium-dose, and high-dose groups were 58%, 40%, and 55%, respectively. The highest number of peritoneal mast cells in mice after in vitro stimulation of degranulation was found in the high-dose group, which also showed the highest tryptase activity. The medium- and high-dose groups, and the positive group exhibited high white blood cell counts (9.7±6.1, 9.6±2.4, and 9.2±2)×109/L, respectively, and the platelet counts were in descending order of low-dose group, medium-dose group, and high-dose group. The results indicated that after sensitization, the coagulation function of mice was significantly impaired, and the immune system was affected; the in vivo production of IgE antibody was induced, and the release mechanism of allergy mediators was activated, leading to the occurrence of anaphylactic reactions (type I hypersensitivity).
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[基金项目]
国家自然科学基金项目(31601421);天津市优秀博士后培养计划项目(2015018);2015年大学生国家创新项目(201510061088)