福多司坦对慢阻肺模型大鼠肺脏及肺血管组织形态和纤维化的影响

Effects of fudosteine on lung and pulmonary vascular histomorphology and fibrosis in chronic obstructive pulmonary disease model rats

目的探究福多司坦对慢阻肺(COPD)模型大鼠肺脏及肺血管组织形态和肺纤维化的影响及可能机制。方法选择18只大鼠并随机分为对照组、模型组、福多司坦组三组。模型组、福多司坦组建立COPD模型(烟熏+脂多糖气管滴注),福多司坦组在烟熏前给予50 kg/mg福多司坦灌胃,模型组给予等量生理盐水。4周后检测各组大鼠肺功能及肺湿重/干重比(W/D),透射电镜观察大鼠肺组织病理变化,维多利亚蓝+VG染色观察肺血管形态变化,免疫荧光染色激光共聚焦显微镜下观察肺纤维化蛋白(ɑ-SMA,E-Cadherin)表达,ELISA检测肺泡灌洗液(BALF)中氧化应激指标(SOD,GSH-PX)和炎症因子(IL-25、IL-17A、TNF-ɑ)水平,Western blot检测肺组织中TGF-β1、Smad3蛋白表达量。结果与对照组比较,模型组、福多司坦组肺组织病理损伤、肺血管炎症浸润及重构、肺纤维化较为明显,大鼠气道流速、跨肺压、W/D、肺纤维化蛋白ɑ-SMA、E-Cadherin表达、BALF中GSH-PX、IL-25、IL-17A、TNF-ɑ含量及肺组织匀浆中TGF-β1、Smad3蛋白表达量升高,SOD、潮气量下降(P<0.05);与模型组比较,福多司坦组肺组织病理损伤、肺血管炎症浸润及重构、肺纤维化程度减轻,大鼠气道流速、跨肺压、W/D、肺纤维化蛋白ɑ-SMA、E-Cadherin表达、BALF中GSH-PX、IL-25、IL-17A、TNF-ɑ含量及肺组织中TGF-β1、Smad3蛋白表达量降低,SOD、潮气量增加(P<0.05)。结论福多司坦能改善COPD大鼠肺部病理、肺血管重塑及纤维化,这可能与其能抑制TGF-β1/Smad通路,减轻氧化应激及炎症损伤有关。

Objective To investigate the effect of fudosteine on lung and pulmonary vascular histomorphology and pulmonary fibrosis in chronic obstructive pulmonary disease(COPD)model rats and its possible mechanism.Methods Eighteen rats were randomly divided into a control group,a model group and a fudosteine group.COPD model was established in the model group and the fudosteine group(smudgingand tracheal drip of lipopolysaccharide).Rats in fudosteine group were given 50 kg/mg fudosteine by gavage before smudging,and rats in model group were given the same amount of saline as fudosteine group.Four weeks later,the pulmonary function and lung wet/dry weight ratio(W/D)of rats in each group were measured.The pathological changes of rat lung tissues were observed under transmission electron microscope,the morphological changes of pulmonary vessels were observed by Victoria blue and Van Gieson collagen fiber staining,the expression of pulmonary fibrosis protein(ɑ-SMA,E-Cadherin)was observed under confocal laser scanning microscope by immunofluorescence staining,the levels of oxidative stress indicators(SOD,GSH-PX)and inflammatory factors(IL-25,IL-17 A,TNF-α)in bronchoalveolar lavage fluid(BALF)were detected by ELISA,and the expression levels of TGF-β1 and Smad3 proteins in lung tissues were detected by Western blot.Results Compared with the control group,the pathological injury of lung tissue,pulmonary vascular inflammatory infiltration and remodeling,and pulmonary fibrosis were more obvious in the model group and fudosteine group.The airway flow velocity,transpulmonary pressure,W/D,pulmonary fibrosis proteinɑ-SMA and E-Cadherin expression,GSH-PX,IL-25,IL-17 A and TNF-αcontents in BALF,and TGF-β1 and Smad3 protein expression in lung homogenates were increased in those two groups;however,the SOD and tidal volume were decreased(P<0.05).Compared with the model group,the pathological injury of lung tissue,pulmonary vascular inflammatory infiltration and remodeling,and the degree of pulmonary fibrosis were alleviated in fudosteine group.The airway flow velocity,transpulmonary pressure,W/D,pulmonary fibrosis protein-SMA,and E-Cadherin expression,GSH-PX IL-25,IL-17A,and TNF-αcontents in BALF and TGF-β1 and Smad 3 protein expression in lung tissue were decreased,SOD AND the tidal volume were increased in the fudosteine group(P<0.05).Conclusions Fudosteine could improve pulmonary pathology,pulmonary vascular remodeling and fibrosis in COPD rats,which may be related to the inhibition of TGF-β1/Smad pathway and the alleviation of oxidative stress and inflammatory injury.