TLR4在慢性支气管炎大鼠模型中对炎症损伤与黏液分泌的调控作用

发布时间：2018-07-20 20:00

【摘要】：目的观察TLR4在细菌内毒素(LPS)致慢性支气管炎大鼠模型中对气道炎症损伤与黏液分泌的调控作用。方法 SPF级雄性Wistar大鼠40只,随机分为正常对照组(NS组)、慢性支气管炎组(LPS组)、多粘菌素B干预组(LPS+PMB组)、多粘菌素B自身对照组(PMB组),每组10只。用气管内注入LPS 200μg/200μl及每日LPS溶液雾化吸入1 h的方法建立慢性支气管炎动物模型,3 w后收集支气管肺泡灌洗液(BALF)进行细胞学计数和白细胞分类检测,酶联免疫吸附实验(ELISA)法测定BALF中肿瘤坏死因子(TNF)-α含量,对大鼠肺脏进行病理学检查,肺组织阿先蓝过碘酸雪夫(AB-PAS)阳染面积,用免疫组化法观察黏蛋白Muc5AC、TLR4在支气管肺内的表达及荧光定量RT-PCR Muc5AC mRNA、TLR4 mRNA在肺内的表达。结果 LPS组大鼠BALF细胞总计数、中性粒细胞个数、TNF-α水平。AB-PAS阳染面积、Muc5AC、TLR4蛋白表达量及Muc5AC mRNA、TLR4 mRNA水平与LPS+PMB组比较有显著性差异(均P0.05),NS组和PMB无统计学差异(P0.05)。结论 TLR4介导LPS诱发的慢性呼吸道炎症反应可能导致Muc5AC蛋白与mRNA高表达。多粘菌素B可能通过抑制肺组织TLR4 mRNA及其蛋白的表达而抑制气道黏液高分泌。
[Abstract]:Objective to observe the effect of TLR4 on airway inflammation injury and mucus secretion in rat model of chronic bronchitis induced by bacterial endotoxin (LPS). Methods Forty SPF male Wistar rats were randomly divided into normal control group (NS group), chronic bronchitis group (LPS group), polymyxin B intervention group (LPS PMB group) and polymyxin B self control group (PMB group). Bronchoalveolar lavage fluid (BALF) was collected from bronchoalveolar lavage fluid (BALF) for 3 weeks after endotracheal injection of LPS 200 渭 g / 200 渭 l and daily LPS solution atomization inhalation for 1 h. The content of tumor necrosis factor (TNF)-伪 in BALF was determined by enzyme-linked immunosorbent assay (Elisa). The expression of mucin Muc5AC TLR4 in bronchopulmonary and the expression of Muc5AC mRNA-TLR4 mRNA in lung were observed by immunohistochemical method. Results the total number of BALF cells, the number of neutrophils and the level of TNF- 伪 in LPS group were significantly higher than those in LPS PMB group (P0.05). There was no significant difference between NS group and PMB group in the expression of Muc5AC mRNATLR4 protein and the level of Muc5AC mRNATLR4 mRNA (P0.05). Conclusion TLR4 mediated LPS-induced chronic respiratory inflammation may lead to the overexpression of Muc5AC protein and mRNA. Polymyxin B may inhibit airway mucus hypersecretion by inhibiting the expression of TLR4 mRNA and its protein in lung tissue.
【作者单位】：宜宾市第一人民医院呼吸内科;遵义医学院附属医院呼吸二科;
【基金】：贵州省科技计划课题(黔科合SY字〔2008〕3056)
【分类号】：R562.21;R-332

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