鼠曲草提取物对COPD模型大鼠气道炎症的抑制作用及对NF-κB通路的影响

Inhibitory effect of Gnaphalium affine extract on inflammation and influence on NF-κB signaling pathway in COPD rats

目的:研究鼠曲草提取物对慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD)模型大鼠气道炎症的影响及作用机制。方法:大鼠随机分为正常组,模型组,鼠曲草提取物低、中、高剂量组(0.75,1.5,3 g·kg^(-1))和地塞米松组(阳性对照药,27 mg·kg^(-1)),每组9只。除正常组外,其余各组大鼠采用香烟烟熏联合气管内滴注脂多糖建立COPD大鼠模型。造模15 d后,各给药组大鼠灌胃相应药物,正常组和模型组大鼠灌胃等体积0.9%氯化钠溶液,每日1次,连续14 d。末次给药后,检测各组大鼠肺功能;HE染色观察肺组织病理学变化;酶联免疫法检测肺泡灌洗液、血清中IL-6、IL-8、IL-17、IL-1β、TNF-α、TGF-β含量;RT-PCR法检测右肺组织中NF-κB mRNA的相对表达量;Western blot法检测右肺组织中NF-κB、IKKβ、IκBα及其p-IκBα蛋白的相对表达量。结果:不同剂量的鼠曲草提取物干预后,均可显著提高大鼠肺功能参数FEV0.3、FVC、FEV0.3/FVC,明显降低肺泡灌洗液中IL-8、IL-17、TNF-α及血清中IL-1β、IL-6、IL-8、IL-17、TNF-α、TGF-β含量,改善大鼠肺组织病理变化,同时显著降低p-IκB、IKK蛋白及NF-κB mRNA和蛋白相对表达量。结论:鼠曲草能够改善COPD模型大鼠的肺功能,减轻肺组织病理损伤,抑制炎症反应,其作用可能与抑制NF-κB信号通路有关。

OBJECTIVE To study the effect and mechanism of Gnaphalium affine extract on inflammation in chronic obstructive pulmonary disease(COPD) rats.METHODS Rats were randomly divided into normal group, model group, low, medium and high dose of Gnaphalium affine extract groups(0.75, 1.5, 3 g·kg^(-1)) and dexamethasone group(positive control drug, 27 mg·kg^(-1)) with 9 rats in each group. Except the normal group, the rats in the other groups were established the COPD model by using cigarette smoking combined with intratracheal lipopolysaccharide. After 15 days of modeling, the rats in each administration group were given the corresponding drug, and the rats in the normal group and the model group were given the same volume of 0.9% NaCl solution once a day for 14 consecutive days. After the last drug administration, the pulmonary function of each group was detected;HE staining was used to observe the pathological changes of lung tissue;the enzyme-linked immunoassay was used to detect the contents of IL-6, IL-8, IL-17, IL-1β, TNF-α and TGF-β in alveolar lavage fluid and serum;real-time PCR method was used to detect the relative expression of NF-κB mRNA in the right lung tissue;Western blot was used to detect the relative expression of NF-κB, IKKβ, IκBα and its phosphorylation(p-IκBα) protein in the right lung tissue.RESULTS The pulmonary function parameters FEV0.3, FVC and FEV0.3/FVC of the rats in each administration group of Gnaphalium affine extract were significantly increased;the levels of IL-8, IL-17 and TNF-α in the bronchoalveolar lavage fluid were significantly reduced, and the levels of IL-1β, IL-6, IL-8, IL-17, TNF-α and TGF-β were significantly reduced;the pathological changes of lung tissue in rats were improved. At the same time, the relative expression levels of p-IκB, IKK protein and NF-κB mRNA and protein in the lung tissues of each administration group of Gnaphalium affine extract were significantly reduced.CONCLUSION Gnaphalium affine extract can effectively improve the lung function of COPD rats, reduce the pathological damage of lung tissue, and reduce the inflammation. Its effects may be related to the inhibition of NF-κB signaling pathway.