中国免疫学杂志Issue(6)：779-783,5.DOI:10.3969/j.issn.1000-484X.2014.06.013
过敏性哮喘模型脱敏治疗前后小鼠肥大细胞的变化与其细胞因子的检测
Observation of mast cells and detection of related cytokines in murine bronchial asthma model pre and post desensitization therapy
摘要
Abstract
Objective:To prepare Dermatophagoides farinae (Der f) crude protein to establish BALB/c bronchial asthma model , and to observe the morphology and degranulation of mast cells and detect related cytokines .Methods: Dermatophagoides farinae ( Der f) crude protein were prepared by trituration .30 BALB/c mice were randomly divided into 3 groups:PBS control group (A), asthma model group (B) and Der f crude protein treatment group (C).Group A were treated with PBS(100 μl) all the time, group B and group C were treated with 50 μg Der f crude protein mixed with 50μl alum adjuvant on day 0,day 7 and day 14.On day 28 group A and B were subcutaneous injected with PBS (100 μl) and group C were subcutaneous injected with Der f crude protein (350μg) in PBS(100 μl) at 1-day intervals.One week after the last treatment ,group A,B and C were intranasally challenged with 50 μg Der f crude protein daily for seven days .Twenty-four hours after the last challenge , airway hyper-responsiveness ( AHR) was assessed by using whole-body plethysmography .Two days post challenged , mice were sacrificed and bronchoalveolar lavage fluid ( BALF) was collected.Number of the total cells and eosinophil was determined .Level of IL-4,IL-10 and IFN-γcytokines in the BALF and the su-pernatant of splenocyte culture was assayed by ELISA .Level of Der f specific IgE and histamine in the sera was determined by ELISA . Airway inflammation was analyzed by HE staining .Observation of the morphology and degranulation of mast cells was analyzed by tolui -dine blue staining.Results:Compared with group B,AHR and the lung inflammation in group C were greatly reduced (P<0.01). Numbers of total cells and eosinophils in BALF of group C were significantly lower than that of group B ( P<0.01 ) .Compared with group B, the observation of degranulation of mast cells was insignificant in group C .Compared with group B(IgE:1.905), the level of specific IgE was significantly lower in groups C (IgE:1.278)(P<0.01).The level of IL-4 in BALF of group C was significantly lower than that of group B(P<0.01).Compared with group A and B, the level of IL-10 in BALF was significantly higher in group C (P<0.01) and the level of IFN-γin BALF of group C was significantly higher than that of group A and B (P<0.01).Compared with group B, the level of IL-4 in cultured splenocytes was significantly lower in group C (P<0.01), and the level of IL-10 and IFN-γin cultured splenocytes of group C was significantly higher than that of group B (P<0.01).Compared with group B, the level of histamine in BALF was slightly lower in groups C (P<0.05), and the level of histamine in sera was significantly lower in groups C (P<0.05). Conclusion:The degranulation of mast cells of murine bronchial asthma model was suppressed after the desensitization therapy .关键词
粉尘螨/过敏性哮喘/肥大细胞/明矾佐剂Key words
Dermatophagoides-farinae/Bronchial-asthma/Mast-cells/Alum-adjuvant分类
医药卫生引用本文复制引用
姚添淇,吴莹莹,杨小猛,匡渤海,刘志刚..过敏性哮喘模型脱敏治疗前后小鼠肥大细胞的变化与其细胞因子的检测[J].中国免疫学杂志,2014,(6):779-783,5.基金项目
本文受海外及港澳学者合作研究基金（No.31328014）、广东省自然科学基金项目（No.04554；S2013040014206）、广东省高等学校国际暨港澳台科技合作创新平台项目（ No.2012gjhz0009）和深圳市科技计划项目（ No.JCYJ20120613100657482、 JCYJ20120613173913654、JCYJ20120613173233810）资助。 （）
