葎草花粉特异性致敏哮喘动物模型的制备

Establishment of allergic mouse asthma model with Humulus pollen allergen

目的建立葎草花粉粗浸液诱导的小鼠变态反应气管炎症动物模型。方法制备葎草花粉粗浸液,将40只清洁级雌性BALB/c小鼠按随机数字表法分为4组,每组10只。①空白对照组:PBS+氢氧化铝致敏激发;②低剂量组:100μg葎草粗浸液+氢氧化铝致敏激发;③中剂量组:200μg葎草粗浸液+氢氧化铝致敏激发;④高剂量组:300μg葎草粗浸液+氢氧化铝致敏激发。通过HE染色观察小鼠肺部炎症和黏液分泌,支气管肺泡灌洗液中细胞总数和细胞分类,酶联免疫吸附试验检测BALF,脾组织匀浆上清的IL-4、IFN-γ和血清抗原特异的抗体验证哮喘动物模型。结果随着抗原浓度的升高,肺部病理改变呈明显的变态反应炎症;高剂量组小鼠支气管肺泡灌洗液中的细胞总数为(210.75±78.80)×106/L、巨噬细胞数为(48.65±3.27)%、EOS数为(24.37±4.28)%、IL-4、血清抗原特异性IgE抗体、IgG1抗体明显高于空白对照组(P<0.05);支气管肺泡灌洗液﹑脾组织匀浆上清的IFN-γ较空白对照组下降(P<0.05)。结论葎草花粉成功构建小鼠变态反应气道炎症动物模型。

Objective To establish the allergic trachitis model of mice with Humulus pollen allergen infusion.Methods Humulus pollen allergen infusion was prepared.Forty clean and female BALB/c mice were randomly divided into blank control group sensitized with PBS ＋ aluminium hydroxide,low dose group sensitized with 100 μg Humulus pollen allergen infusion ＋ aluminium hydroxide,medium dose group sensitized with 200 μg Humulus pollen allergen infusion ＋ aluminium hydroxide,and high dose group sensitized with 300 μg Humulus pollen allergen infusion ＋ aluminium hydroxide,10 mice in each group.After lung tissue samples from mice were stained with haematoxylin and eosin（HE）,inflammation and mucus secretion were observed,total number of cells in alveoli lavage solution and their composition were calculated,BALF,IL-4,IFN-γ,IgE and IgG1 in homogenate supernatant of splenocytes were assayed by enzyme-linked immunoassay（ELISA）.Results Allergic inflammation aggravated with the increasing concentration of allergen.The total number of cells,macrophages,and EOS in alveoli lavage solution was（210.75 ± 78.80） × 106 /L,48.65% ± 3.27%,and 24.37% ± 4.28%,respectively.The serum levels of IL-4,IgG1 and IgE were significantly higher in low,medium and high dose groups than in blank control group（P 0.05）.However,the IFN-γ level in alveoli lavage solution and homogenate supernatant of splenocytes was lower in low,medium and high dose groups than in blank control group（P 0.05）.Conclusion Humulus pollen allergen infusion can be used in establishing the allergic trachitis model of mice.