和解抗敏煎对呼吸道合胞病毒感染诱发哮喘小鼠TLR3及NF-κB/IRF3信号通路调控的研究

Study on the Regulation of Conciliatory Antiallergic Decoction on TLR3 and NF-κB/IRF3 Signaling Pathway in Asthmatic Mice Induced by Respiratory Syncytial Virus

目的探讨和解抗敏煎对呼吸道合胞病毒(respiratory syncytial virus,RSV)感染诱发哮喘小鼠TLR3及NF-κB/IRF3信号通路的调控作用。方法将BALB/c小鼠随机分为对照组,模型组,地塞米松组(0.5 mg·kg^-1),和解抗敏煎低、中、高剂量组(17.5,35.0,70.0 g·kg^-1),每组10只。RSV诱发合并卵白蛋白致敏建立哮喘小鼠模型。末次激发后24 h,乙酰甲胆碱吸入测定气道相对阻力;麻醉小鼠,制备肺泡灌洗液,检测炎症细胞数(巨噬细胞、嗜酸性粒细胞、淋巴细胞、中性粒细胞)和IgE、CD31、IL-4、TNF-α、MPO含量。取肺组织石蜡包埋切片行HE染色观察肺组织炎症浸润,免疫组化染色观察TLR3表达。qRT-PCR检测肺组织TLR3、NF-κB、IRF3 mRNA的表达。结果与模型组相比,和解抗敏煎各剂量组能有效降低哮喘小鼠气道高反应性,改善肺组织损伤和炎症细胞浸润,降低肺泡灌洗液中巨噬细胞、嗜酸性粒细胞、淋巴细胞、中性粒细胞数量和IgE、CD31、IL-4、TNF-α、MPO的含量,并有效降低肺组织TLR3、NF-κB、IRF3的表达。结论和解抗敏煎对病毒诱发的支气管哮喘具有治疗效果,且可能与介导TLR3及NF-κB/IRF3信号通路有关。

OBJECTIVE To explore the regulation effect of conciliatory antiallergic decoction on TLR3 and NF-κB/IRF3 signaling pathway in allergic asthma mice induced by respiratory syncytial virus(RSV). METHODS BALB/c mice were randomly divided into control group, model group, dexamethasone group(0.5 mg·kg^-1), conciliatory antiallergic decoction low, medium and high dose groups(17.5, 35.0, and 70.0 g·kg^-1), with 10 mice in each group. Asthma model was established by RVS combined with ovalbumin sensitization. Twenty-four hours after the last challenge, airway resistance was examined via mice exposing to vaporized methacholine. Mice was anesthetized and broncho-alveolar lavage fluid was prepared. Then the inflammatory cells including macrophages, eosinophils, lymphocytes and neutrophils were counted, the contents of IgE, CD31, IL-4, TNF-α and MPO were detected. Lung tissues injury were evaluated with HE staining, TLR3 expression in lung tissues was determined with immunohistochemical staining, and the mRNA expression of TLR3, NF-κB and IRF3 in lung tissues were detected with qRT-PCR. RESULTS Compared with the model group, the airway resistance in all dose of conciliatory antiallergic decoction groups were significantly inhibited, the lung tissues injury and inflammatory cell infiltration were also attenuated, the counts of macrophages, eosinophils, lymphocytes and neutrophils were significantly decreased, the contents of IgE, CD31, IL-4, TNF-α and MPO in broncho-alveolar lavage fluid were also decreased. The expression of TLR3, NF-κB and IRF3 in lung tissues were also decreased. CONCLUSION Conciliatory antiallergic decoction has treatment effect against asthma induced by virus, this may related to the regulation of TLR3/NF-κB/IRF3 signaling pathway.