蒲公英总黄酮对烟草烟雾致慢性阻塞性肺疾病的抗氧化保护作用及机制  

Antioxidant Protective Effect and Mechanism of Total Flavonoids from Dandelion on Cigarette Smoke-Induced Chronic Obstructive Pulmonary Disease

作　　者：王超 陈芳[1] 胡凯淞 安一鸣 郑文学 张嘉琪 陈楷[2] 王国强[1] 郑敬彤[1] 刘金平[3] 王放[1] Wang Chao;Chen Fang;Hu Kaisong;An Yiming;Zheng Wenxue;Zhang Jiaqi;Chen Kai;Wang Guoqiang;Zheng Jingtong;Liu Jinping;Wang Fang(College of Basic Medical Sciences,Jilin University,Changchun 130021;Hospital of Stomatology,Jilin University,Changchun 130021;School of Pharmaceutical Sciences,Jilin University,Changchun 130021)

机构地区：[1]吉林大学基础医学院,长春130021 [2]吉林大学白求恩口腔医学院,长春130021 [3]吉林大学药学院,长春130021

出　　处：《国际老年医学杂志》2023年第3期257-264,共8页International Journal of Geriatrics

基　　金：吉林省预算内基本建设基金(20119C030);吉林省预算内基本建设基金(2022C036);吉林省科技发展计划基金(20210502005ZP)。

摘　　要：目的探讨蒲公英总黄酮对烟草烟雾(CS)致慢性阻塞性肺疾病(COPD)模型小鼠的影响,并阐明其相关作用机制。方法首先,基于DPPH和ABTS方法检测蒲公英茎叶总黄酮提取物和根总黄酮提取物的体外抗氧化能力。其次,将60只雌性SPF级BALB/c小鼠随机分为空白对照(Con)组、CS组、CS-根总黄酮低剂量(Root-LD)组、CS-茎叶总黄酮低剂量(Leaf-LD)组、CS-根总黄酮高剂量(Root-HD)组和CS-茎叶总黄酮高剂量(Leaf-HD)组,COPD模型制备采用被动吸烟法,每次4根香烟,每天累积烟熏时长为1 h,持续造模12周。治疗组小鼠在每天烟熏前2 h予以灌胃给药治疗,Root-LD组剂量为2.25 g/kg,Root-HD组为4.50 g/kg,Leaf-LD组为2.25 g/kg,Leaf-HD组为4.50 g/kg。最后,基于小鼠肺功能仪检测各组的肺功能相关指标,包括气道阻力(Raw)、动态肺顺应性(Cdyn)及第0.1秒用力呼气容积(FEV0.1)等指标;基于肺组织的HE染色结果比较各组小鼠肺部的病理变化;使用试剂盒检测各组肺组织中丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽(GSH)的水平;使用RT-PCR和免疫组织化学方法分别检测核因子红细胞2相关因子2(Nrf2)信号通路相关基因和蛋白的表达。结果蒲公英茎叶总黄酮和根总黄酮均具有良好的抗氧化能力,DPPH方法的EC50值分别为54.88μg/mL、123.50μg/mL,ABTS方法的EC50值分别为229.41μg/mL、559.07μg/mL。蒲公英总黄酮治疗后可以显著恢复COPD小鼠的体质量(P<0.05);蒲公英的茎叶总黄酮提取物能显著改善COPD小鼠的Cdyn、FEV0.1及Raw等肺功能指标(P<0.05),且茎叶总黄酮提取物的效果要优于根部总黄酮提取物;HE染色结果表明CS诱导的COPD小鼠肺部病理变化符合临床慢性气道炎症的病理表现,并且蒲公英总黄酮能显著抑制CS诱导的COPD小鼠模型的炎症反应;蒲公英总黄酮可以上调抗氧化基因Nrf2和SOD1的mRNA水平;免疫组织化学结果表明蒲公英总黄酮可以调节Nrf2信号通路的相关Objective To investigate the effects of total flavonoids from Dandelion on mice with chronic obstructive pulmonary disease(COPD)induced by cigarette smoke(CS)and to elucidate the mechanisms involved.Methods The antioxidant capacity of total flavonoid extracts from Dandelion stems-and-leaves,and roots were examined using DPPH and ABTS.Sixty female SPF-grade BALB/c mice were randomly divided into a blank control(Con)group,CS group,CS-root total flavonoid low dose(Root-LD)group,CS-leaf total flavonoid low dose(Leaf-LD)group,CS-root total flavonoid high dose(Root-HD)group,and CS-leaf total flavonoid high dose(Leaf-HD)group.COPD model was prepared by passive smoking method,with 4 cigarettes each time,and the cumulative duration of smoking was one hour per day for 12 weeks.The mice in the treatment group were treated by gavage two hours before smoking each day at a dose of 2.25 g/kg for Root-LD group,4.5 g/kg for Root-HD group,2.25 g/kg for Leaf-LD group and 4.5 g/kg for Leaf-HD group.The lung function-related indices were examined,including airway resistance(Raw),dynamic compliance(Cdyn),and forced expiratory volume in 100 ms(FEV0.1).The pathological changes of the lung tissues were examined by HE staining.The levels of malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione(GSH)in the lung tissues of mice were measured using kits.The expression levels of genes and proteins related to Nrf2 signaling pathway were detected using RT-qPCR and immunohistochemical staining,respectively.Results Both total flavonoids from Dandelion stems-and-leaves and total flavonoids from roots showed good antioxidant capacity with EC50 values of 54.88μg/mL and 123.50μg/mL for DPPH,and 229.41μg/mL and 559.07μg/mL for ABTS.The total flavonoids from Dandelion significantly restored body weight of COPD mice(P<0.05).The total flavonoid from Dandelion leaves significantly improved lung function indices including Cdyn,FEV0.1 and Raw in COPD mice,the differences were significant(P<0.05),and the effect of the total flavonoid from leaves

关 键 词：蒲公英总黄酮 慢性阻塞性肺疾病 抗氧化 肺功能 核因子红细胞2相关因子2 

分 类 号：R285.5[医药卫生—中药学]