摘要
目的:建立大鼠慢性阻塞性肺疾病(COPD)模型,探讨葛根素是否能够通过调控PTEN诱导假定激酶1(PINK1)-parkin信号通路介导的线粒体自噬,发挥肺保护作用。方法:气管滴注脂多糖+烟熏法建立COPD大鼠模型,建模成功后随机分为模型组、低剂量(50 mg/kg)葛根素组、中剂量(100 mg/kg)葛根素组、高剂量(200 mg/kg)葛根素组和3-甲基腺嘌呤(3-MA;10 mg/kg)组,每组15只;另取15只大鼠作为对照组。连续给药8周后进行大鼠肺功能检测;ELISA法检测大鼠肺泡灌洗液中TNF-α、IL-1β和IL-6水平;HE染色观察大鼠肺组织病理变化;TUNEL染色观察大鼠肺泡上皮细胞凋亡情况;免疫组化法检测肺组织中BECN1和LC3B蛋白表达;Western blot法检测肺组织中PINK1、parkin、Bax和Bcl-2蛋白表达。结果:与对照组比较,模型组大鼠肺组织病变明显,肺泡腔严重充血,炎症细胞大量浸润,肺泡间隔与肺泡壁增厚,病理评分显著升高(P<0.05),用力肺活量(FVC)、0.1 s用力呼气容积(FEV0.1)和呼气流量峰值(PEF)水平均显著下降(P<0.05),TNF-α、IL-1β和IL-6水平,肺泡上皮细胞凋亡率,肺组织中BECN1、LC3B、Bax、PINK1和parkin表达均显著升高(P<0.05),Bcl-2表达显著降低(P<0.05);与模型组比较,各剂量葛根素组和3-MA组大鼠肺组织病理评分均显著降低(P<0.05),FVC、FEV0.1和PEF水平均显著升高(P<0.05),TNF-α、IL-1β和IL-6水平,肺泡上皮细胞凋亡率,肺组织中BECN1、LC3B、Bax、PINK1和parkin表达均显著降低(P<0.05),Bcl-2表达显著升高(P<0.05),且具有剂量依赖性(P<0.05);与高剂量葛根素组比较,3-MA组大鼠上述指标无显著差异(P>0.05)。结论:葛根素可能通过调控PINK1-parkin信号通路介导的线粒体自噬,抑制肺泡上皮细胞凋亡和肺部炎症反应,从而减轻COPD大鼠肺损伤。
AIM:To establish a rat model of chronic obstructive pulmonary disease(COPD),and to investigate whether puerarin can exert lung protective effect on this model through PTEN-induced putative kinase 1(PINK1)-parkin signaling pathway.METHODS:A COPD rat model was established by intratracheal dripping of lipopolysaccharide+fumigation method.After successful modeling,the rats were randomly divided into model group,low-dose(50 mg/kg),medium-dose(100 mg/kg)and high-dose(200 mg/kg)puerarin groups,and 3-methyladenine(3-MA;10 mg/kg)group,with 15 rats in each group.Another 15 normal rats served as control group.The lung function of the rats was measured after 8 weeks of treatments.The levels of TNF-α,IL-1βand IL-6 in alveolar lavage fluid were detected by ELISA.The pathological changes of the lung tissue were observed by HE staining.The apoptosis of alveolar epithelial cells was observed by TUNEL staining.The protein expression of BECN1 and LC3B in the lung tissue was detected by immunohistochemistry.The protein levels of PINK1,parkin,Bax and Bcl-2 in the lung tissue were detected by Western blot.RESULTS:The lung tissue lesions in model group were observed,with severe congestion in the alveolar cavity,massive infiltration of inflammatory cells,thickening of the alveolar septum and alveolar wall,increased pathological scores(P<0.05),decreased forced vital capacity(FVC),force expiratory volume in 0.1 s(FEV0.1)and peak expiratory flow(PEF)(P<0.05),increased TNF-α,IL-1βand IL-6 levels,alveolar epithelial cell apoptosis rate,and BECN1,LC3B,Bax,PINK1 and parkin expression in the lung tissue,while Bcl-2 expression was significantly decreased(P<0.05).Compared with model group,the lung histopathological scores of rats in puerarin groups and 3-MA group were significantly decreased(P<0.05),FVC,FEV0.1and PEF levels were significantly increased(P<0.05),TNF-α,IL-1βand IL-6 levels,alveolar epithelial cell apoptosis rate,and BECN1,LC3B,Bax,PINK1 and parkin expression in the lung tissue were significantly decreased(P<0.05),while Bcl-2 e
作者
潘晓杰
范嘉铨
夏光琴
PAN Xiao-jie;FAN Jia-quan;XIA Guang-qin(Department of Respiratory and Critical Care Medicine,Jinyang Hospital Affiliated to Guizhou Medical University,Guiyang 550023,China;Department of Respiratory and Critical Care Medicine,The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2022年第10期1833-1839,共7页
Chinese Journal of Pathophysiology
基金
贵州省卫健委科学技术基金项目(gzwjk-2020-1-056号)。
