组织因子对卵清蛋白诱导的小鼠哮喘气道高反应性和炎症的作用

Effects of tissue factor on ovalbumin-induced airway hyperrespon⁃siveness and inflammation in mice with asthma

目的探讨组织因子(TF)对卵清蛋白(OVA)诱导的小鼠气道高反应性(AHR)和炎症的影响。方法选取2020年7月至12月本院哮喘患儿22例,健康对照20例,免疫印迹法和qPCR检测外周血单个核细胞(PBMCs)中TF的表达,酶联免疫法测定(ELISA)血浆中TF表达水平。小鼠随机分为阴性对照(NC)组、OVA组和OVA+shTF组,通过腹腔注射卵清蛋白(OVA)诱导哮喘模型。测量肺阻力指数(RI)和动态顺应性(Cdyn)以确定气道高反应性(AHR),进行苏木精和伊红(HE)染色并评分以显示组织病理学变化,分别测量支气管肺泡灌洗液(BALF)的细胞计数,通过ELISA分析BALF细胞因子白细胞介素(IL)-4、IL-5、IL-13和肿瘤坏死因子α(TNF-α)以及血清中免疫球蛋白E(IgE)水平。结果与健康对照者相比,哮喘患儿PBMCs中TF蛋白和mRNA表达水平升高(P<0.05),血浆中TF蛋白水平也升高(P<0.05)。同样,与NC组小鼠相比,TF表达在OVA诱导的哮喘小鼠中升高,而shTF处理后的哮喘小鼠中TF表达降低(P<0.05)。shTF可逆转OVA诱导的小鼠AHR(P<0.05),减轻病理性炎症浸润,降低血清IgE水平及BALF中细胞因子IL-4、IL-5、IL-13和TNF-α的表达(P<0.05)。此外,shTF降低了OVA诱导的哮喘小鼠BALF中WBC、中性粒细胞、单核细胞和嗜酸性粒细胞的数量(P<0.05)。结论shTF可改善过敏性哮喘小鼠模型的肺功能,减少浸润性炎症细胞。

This study was performed to investigate the effect of tissue factor(TF)on airway hyperresponsiveness(AHR)and inflammation induced by ovalbumin(OVA)in mice.From July to December 2020,total of 22 children with asthma in our hospital and 20 healthy controls were selected.Western blot and qPCR were used to detect the expression of TF in peripheral blood mononuclear cells(PBMCs),and enzyme linked immunosorbent assay(ELISA)was used to detect the expression of TF in plasma.Mice were randomly divided into negative control(NC)group,OVA group and OVA+shTF group,and lung resistance index(RI)and dynamic compliance(Cdyn)were measured to determine airway hyperresponsiveness(AHR),while hematoxylin and eosin(HE)staining and scoring were performed to show histopathological changes.The cell counts of bronchoalveolar lavage fluid(BALF)were measured respectively.The levels of interleukin(IL)-4,IL-5,IL-13,tumor necrosis factor-α(TNF-α)and serum immunoglobulin E(IgE)from BALF were determined by ELISA.The results showed that compared with healthy controls,the expression levels of TF protein and mRNA in PBMCs of children with asthma increased(P<0.05),and the level of TF protein in plasma also increased(P<0.05).Similarly,compared with the NC group mice,TF expression was increased in OVA-induced asthmatic mice,while TF expression was decreased in asthmatic mice after shTF treatment(P<0.05).shTF reversed OVA-induced AHR in mice(P<0.05),reduced pathological inflammatory infiltration,reduced serum IgE levels and the expression of cytokines IL-4,IL-5,IL-13 and TNF-αin BALF(P<0.05).In addition,shTF reduced the number of WBC,neutrophils,monocytes and eosinophils in BALF of OVA-induced asthmatic mice(P<0.05).Taken together,shTF can improve lung function in mouse models of allergic asthma and reduce infiltrating inflammatory cells.