过敏性哮喘模型小鼠肥大细胞的活化及细胞因子的变化  

Activation of Mast Cells and Changes in Cytokines in Mouse Allergic Asthma Model

作　　者：姚添淇 吴莹莹[1] 杨小猛[1] 匡渤海[2] 刘志刚[1] 

机构地区：[1]深圳大学过敏反应与免疫学研究所,广东深圳518060 [2]南昌大学基础医学院遗传教研室,南昌330006

出　　处：《南昌大学学报（医学版）》2014年第4期4-8,107,共5页Journal of Nanchang University：Medical Sciences

基　　金：海外及港澳学者合作研究基金(31328014);广东省自然科学基金(04554);广东省高等学校国际暨港澳台科技合作创新平台项目(2012gjhz0009);深圳市科技计划项目(JCYJ20120613100657482;JCYJ20120613173913654;JCYJ20120613173233810)

摘　　要：目的观察粉尘螨粗抗原建立的Bal b/c哮喘模型小鼠肥大细胞的活化及细胞因子的变化。方法将20只Bal b/c小鼠按随机数字表法分为2组:阴性对照组(A组,n=10),使用100μL PBS处理;哮喘模型组(B组,n=10),50μg粉尘螨粗抗原+50μL明矾佐剂分别在第0、7和14d腹腔注射致敏1次,第28天再用50μg粉尘螨抗原滴鼻激发,每天1次,连续7次。末次激发24h测定气道高反应;48h处死小鼠进行眼球取血,进行肺泡灌洗并收集右肺的肺泡灌洗液(BALF),无菌取肺HE染色病理观察、甲苯胺蓝染色观察肥大细胞脱颗粒,制取脾细胞及粉尘螨粗抗原刺激培养。采用ELISA法测定BALF和脾细胞培养上清IL-4、IL-10和IFN-γ以及血清IgE、组胺抗体含量。结果与A组相比:B组气道高反应性和肺部病理显著加重(P<0.01);BALF细胞总数和嗜酸粒细胞数显著增多(P<0.01);肥大细胞脱颗粒现象显著加重;血清抗原特异性IgE水平显著升高(P<0.01);BALF和脾细胞培养上清IL-4和IL-10水平显著升高(P<0.05或P<0.01),IFN-γ水平显著降低(P<0.01);BALF和血清组胺水平显著升高(P<0.05或P<0.01)。结论过敏性哮喘模型中肥大细胞被活化。Objective To observe the activation of mast cells and changes in cytokines in a Bal b/c mouse asthma model established by Dermatophagoides farinae（Der f）crude antigen.Methods Twenty Bal b/c mice were randomly divided into two groups.Mice in group A were treated with 100μL PBS.However,mice in group B were intraperitoneally injected with 50μg Der f crude an-tigen and 50μL alum adjuvant on day 0,7 and 14 for sensitization,and were intranasally chal-lenged with 50μg Der f antigen for 7 days once per day from day 28.Airway hyperresponsiveness （AHR）was determined 24 hours after the last challenge.Mice were sacrificed and blood samples were taken 48 hours after the last challenge.The right lung bronchoalveolar lavage was performed and bronchoalveolar lavage fluid（BALF）was collected.Pathological changes in the lung were ob-served by HE staining and degranulation of mast cells was analyzed by toluidine blue staining.In addition,spleen cells were separated,cultured and stimulated by Der f crude antigen.Levels of IL-4,IL-10 and IFN-γin BALF and spleen cell culture supernatant and levels of IgE and histamine antibody were determined by ELISA.Results Compared with group A,the AHR,lung inflamma-tion,total cell number in BALF,number of eosinophilic granulocytes in BALF,degranulation of mast cells,antigen-specific serum IgE amount,IL-4 and IL-10 levels in BALF and spleen cell cul-ture supernatant and histamine levels in BALF and serum significantly increased,but IFN-γlevels in BALF and spleen cell culture supernatant obviously decreased in group B（P〈0.05 or P〈0.01）.Conclusion Mast cells were activated in the mouse allergic asthma model.

关 键 词：粉尘螨 过敏性哮喘 肥大细胞 明矾佐剂 动物 实验 小鼠 

分 类 号：R365[医药卫生—病理学]