TNF-α对哮喘小鼠肺泡灌洗液中白三烯含量的影响

摘要

目的研究肿瘤坏死因子(TNF)-α对哮喘小鼠支气管肺泡灌洗液(BALF)中白三烯D4(LTD4)和E4(LTE4)表达的影响,从而探讨其在哮喘发病过程中作用机制。方法48只雌性BALB/c小鼠随机均分为6组：正常对照(A)组,哮喘模型(B)组,TNF-α(0.1μg/kg)腹腔注射给药(C1)组, TNF-α(0.4μg/kg)腹腔注射给药(C2)组,TNF-α(0.1μg/kg)雾化吸入给药(D1)组,TNF-α(0.4μg/kg)雾化吸入给药(D2)组。建立哮喘小鼠模型,苏木精-伊红(HE)染色观察肺组织病理变化,收集BALF 进行炎性细胞计数,酶联免疫疫吸附(ELISA)法检测其 LTD4和 LTE4水平。结果①HE染色显示B、C、D组气道炎症明显改变,C、D组BALF中炎性细胞计数、嗜酸性粒细胞( EOS)明显高于 B 组( P <0.05)；于 ELISA法结果显示BALF中LTD4和LTE4水平：　　B、C、D组高于A组(P<0.01),C、D组明显高于B组(P<0.05),C2组明显高于C1组(P<0.05),D2组明显高于D1组(P<0.05),但C1组和D1组之间、C2和D2组之间比较,差异无统计学意义。结论TNF-α可引起肺内炎性介质LTD4和LTE4产生增加,同时引起EOS和其他炎性细胞在气道和肺组织局部募集、浸润,维持和加重了哮喘病理改变,这可能是哮喘炎症状态持续存在的发生机制之一。%Objective To investigate the expression of leukotriene D4 and leukotriene E4 in the bronchoalveolar lavage fluid(BALF) in asthma mice model and to explore the role of tumor necrosis factor α(TNF-α) played in asthma pathogenesis. Methods Forty-eight BALB/c mice were randomly assigned into normal control(A group), asthma(B group), inhaled low dose TNF-α(C1 group),inhaled high dose TNF-α(C2 group),injected low dose TNF-α(D1 group)and injected high dose TNF-α groups(D2 group)(n=8 each). The mice were sensitized with ovalbumin to establish the asthmatic model. The histological changes were evaluated by hematoxylin and eosin stai-ning, BALF were collected, the total cell and the cell differentials were counted, and the levels of leukotriene D4 and leukotriene E4 were measured by enzyme-linked immunosorbent assay. SPSS 17.0 software was used to analyze the data. Results ① Typical inflammatory changes were shown in asthma group and TNF-α intervention groups. The total cells, EOS amounts of TNF-αintervention group were significantly higher than those of asthma group. ②The expression of LTD4 and LTE4 in BALF:B group,C group and D group were significantly higher than that in A group(P<0.01), C group and D group were significantly higher than B group(P<0.05), C2 group were signifi-cantly higher than C1 group(P<0.05), D2 group were significantly higher than D1 group(P<0.05), but no sta-tistical significance were obtained between C1 group and D1 group, C2 group and D2 group. Conclusion TNF-αcan significantly increase the expression of LTD4 and LTE4 in BALF in asthmatic mice, promote asthma attack and deteriorate airway inflammation. It may be one of the mechanism of TNF-α involved in asthma.