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

发布时间：2018-10-11 18:50

【摘要】：目的:制备粉尘螨粗抗原建立BALB/c小鼠哮喘模型,检测细胞因子并观察肥大细胞形态及脱颗粒的情况。方法:制备粉尘螨粗抗原,30只BALB/c小鼠随机均分为3组,阴性对照组(A组)、哮喘模型组(B组)和治疗组(C组)。A组始终使用PBS处理,B组和C组在第0、7和14天用50μg粉尘螨粗抗原+50μl明矾佐剂腹腔注射致敏。在第28天,A组和B组用PBS、C组用粉尘螨粗抗原(350μg/次)皮下注射免疫1周,隔天免疫1次,末次免疫1周后开始用50μg粉尘螨粗抗原滴鼻激发,每天1次,连续7次。末次激发24 h后进行气道高反应检测。末次激发48 h后处死小鼠取血,对小鼠进行肺泡灌洗收集肺泡灌洗液(BALF),无菌摘取肺组织和脾脏,分别进行肺组织病理切片与脾细胞培养。检测BALF和脾细胞上清中IL-4、IL-10和IFN-γ细胞因子水平,血清中IgE、组胺抗体水平,进行HE染色和甲苯胺蓝染色观察肺部炎症细胞浸润程度和肥大细胞脱颗粒状况。结果:与B组相比,C组气道高反应性和肺部病理症状减轻(P0.01)。C组BALF中细胞计数结果显示与B组相比细胞总数和嗜酸粒细胞数明显减少(P0.01)。与B组相比,C组的肥大细胞脱颗粒情况明显减轻,细胞形态稳定。与B组(IgE:1.905)相比,C组血清中抗原特异性IgE抗体水平(IgE:1.278)明显降低(P0.01)。BALF细胞因子IL-4、IL-10和IFN-γ水平检测结果显示,与B组相比,C组BALF中IL-4水平明显降低接近A组值(P0.01),而IL-10与A、B两组相比有明显升高(P0.01),C组的IFN-γ也高于A、B两组(P0.01)。C组脾细胞上清中IL-4明显低于B组(P0.01),而IL-10明显高于B组(P0.01),IFN-γ也明显高于B组(P0.01)。BALF和血清中组胺水平结果显示,在BALF中,C组与B组相比略低(P0.05),而在血清中检测出的组胺有明显降低(P0.05)。结论:过敏性哮喘模型脱敏治疗后小鼠肥大细胞脱颗粒情况被抑制。
[Abstract]:Aim: to establish asthma model of BALB/c mice with coarse antigen of Dermatophagoides farinae, detect cytokines and observe the morphology and degranulation of mast cells. Methods: thirty BALB/c mice were randomly divided into 3 groups. Negative control group (group A), asthma model group (group B) and treatment group (group C). A) were treated with PBS all the time. Group B and C were sensitized with 50 渭 g adjuvant 50 渭 l of coarse antigen of Dermatophagoides farinae on the 7th and 14th day after treatment. On the 28th day, group A and group B were immunized subcutaneously with the crude antigen of Dermatophagoides farinae (350 渭 g / time) for 1 week in PBS,C group, once every other day. After the last immunization, 50 渭 g crude antigen of Dermatophagoides farinae was given nasal drip once a day for 7 consecutive times. Airway hyperresponsiveness was detected 24 h after the last excitation. The mice were sacrificed for 48 h after the last stimulation, and the alveolar lavage fluid (BALF),) was collected by alveolar lavage. The lung tissue and spleen were removed with (BALF), and the pathological sections of lung tissue and the culture of spleen cells were carried out respectively. The levels of IL-4,IL-10 and IFN- 纬 cytokines in the supernatant of BALF and spleen cells, and the level of IgE, histamine antibody in serum were measured. The infiltration of inflammatory cells and degranulation of mast cells were observed by HE staining and toluidine blue staining. Results: compared with group B, the airway hyperresponsiveness and lung pathological symptoms were alleviated in group C (P0.01). C group). The cell count in BALF of group C was significantly lower than that in group B (P0.01), and the total number of cells and the number of eosinophils in group B were significantly lower than those in group B (P0.01). Compared with group B, the degranulation of mast cells in group C was significantly reduced and cell morphology was stable. Compared with group B (IgE:1.905), the level of antigen-specific IgE antibody (IgE:1.278) in serum of group C was significantly lower than that of group B (P0.01). BALF cytokines IL-4,IL-10 and IFN- 纬). Compared with group B, the level of IL-4 in BALF in group C was significantly lower than that in group A (P0.01), while the level of IFN- 纬 in group IL-10 was significantly higher than that in group A and B (P0.01), C group was also higher than that in group A and B group (P0.01). C group was significantly lower than that in group B (P0.01), IL-10 was significantly higher than group B (P0.01), IFN- 纬 was significantly higher than group B (P0.01). It was also significantly higher than that in group B (P0.01). BALF and serum histamine level). In BALF, group C was slightly lower than group B (P0.05), while histamine detected in serum was significantly lower (P0.05). Conclusion: the degranulation of mast cells was inhibited after desensitization in allergic asthma model.
【作者单位】：深圳大学过敏反应与免疫学研究所;南昌大学医学院遗传教研室;
【基金】：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) 深圳市科技计划项目(No.JCYJ20120613100657482、JCYJ20120613173913654、JCYJ20120613173233810)资助
【分类号】：R562.25

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