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目的:研究特异性免疫治疗(SIT)对哮喘小鼠自然杀伤T(NKT)细胞的影响。方法:24只BALB/C小鼠随机分为对照组(A组)、哮喘模型组(B组)、哮喘免疫治疗(SIT)组(C组),各8只。通过屋尘螨提取液(HDM)诱导建立哮喘小鼠模型并进行SIT治疗。检测各组小鼠的气道反应性、支气管肺泡灌洗液(BAlF)细胞计数及分类、ELISA检测IL-4、IFN-γ以及应用流式细胞仪检测NKT细胞数目,通过RT-PCR方法检测T-bet和GATA-3mRNA表达水平;HE染色观察小鼠肺组织的改变。结果:与B组相比,C组气道反应性明显下降(P<0.01);BALF中细胞总数及嗜酸性粒细胞(EOS)数显著减少(P<0.01);血清IL-4分泌显著降低(P<0.01),IFN-γ显著升高(P<0.01);NKT细胞数及其成熟型比例明显升高(P<0.05);T-betmRNA表达水平明显升高(P<0.01),且与NKT细胞数及其成熟型比例呈正相关性,GATA-3mRNA表达水平明显降低(P<0.05),且与NKT细胞数及其成熟型比例呈负相关性。B组肺部管腔周围炎性细胞聚集,组织上皮损伤,组织水肿,而C组肺部变应性炎症明显减轻。C组其他各项指标接近A组。结论:哮喘的发生可能与NKT细胞失调相关,通过改变NKT细胞数目及其成熟型比例来调节GATA-3/T-bet的表达可能是SIT治疗哮喘的作用机制之一。
Objective: To investigate the effect of specific immunotherapy (SIT) on natural killer T (NKT) cells in asthmatic mice. Methods: Twenty-four BALB / C mice were randomly divided into control group (A group), asthma model group (B group) and asthma immunization (SIT group) (C group). The mouse model of asthma was induced by house dust mite extract (HDM) and treated with SIT. The airway responsiveness, bronchoalveolar lavage fluid (BAlF) cell count and classification, IL-4 and IFN-γ by ELISA and NKT cell number by flow cytometry were detected by RT-PCR T-bet and GATA-3mRNA expression levels were observed; HE staining was used to observe the changes of lung tissue in mice. Results: Compared with group B, the airway reactivity of group C was significantly decreased (P <0.01); the number of total cells and eosinophils (EOS) in BALF were significantly decreased (P <0.01); the level of IL-4 secretion was significantly decreased (P <0.01), while the level of IFN-γ was significantly increased (P <0.01). The number of NKT cells and its mature type were significantly increased (P <0.05), and the expression of T-bet mRNA was significantly increased It was positively correlated with the number of NKT cells and its mature type. The expression level of GATA-3 mRNA was significantly decreased (P <0.05), and negatively correlated with the number of NKT cells and the percentage of mature type. In Group B, the inflammatory cells around the luminal cavity were aggregated, the epithelial tissue was damaged and the tissue was edematous. The allergic inflammation in the lung of Group C was significantly reduced. Other indicators of group C are close to group A. CONCLUSIONS: The development of asthma may be related to the imbalance of NKT cells. Regulation of GATA-3 / T-bet expression by changing the number of NKT cells and their mature proportion may be one of the mechanisms of SIT in treating asthma.