槲皮素预防急性加重慢性阻塞性肺疾病的作用及机制研究  

The Protective Effects and Mechanisms of Quercetin on Acute Exacerbation of Chronic Obstructive Pulmonary Disease

作　　者：张鹏 叶陈丽[1] 马婧 曹伟灵[1] ZHANG Peng;YE Chenli;MA Jing;CAO Weiling(Department of Pharmacy,Shenzhen Luohu People's Hospital,Shenzhen,Guangdong 518001,China;The Eighth Hospital,Sun Yat-sen University,Shenzhen,Guangdong 518000,China)

机构地区：[1]深圳市罗湖区人民医院药学部,广东深圳518001 [2]中山大学附属第八医院,广东深圳518000

出　　处：《今日药学》2021年第6期416-421,共6页Pharmacy Today

摘　　要：目的探讨槲皮素对急性加重慢性阻塞性肺疾病(AECOPD)的预防作用及相关机制,为槲皮素的临床应用提供理论依据。方法采用香烟烟雾提取液(CSM)及脂多糖(LPS)联合刺激人支气管上皮BEAS-2B细胞,构建AECOPD体外细胞模型。采用槲皮素(10μmol·L-1)预处理细胞1 h后,加入CSM(4%,v/v)和LPS(10μg·mL-1)继续培养细胞24 h,收集细胞及上清液。利用酶联免疫法检测培养液中白介素-8(IL-8)的含量,硫代巴比妥酸反应物法检测细胞内丙二醛(MDA)的水平,试剂盒检测细胞内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、磷酸二酯酶4(PDE-4)、环磷酸腺苷(cAMP)及蛋白激酶A(PKA)的活性,免疫蛋白印迹法检测细胞中核因子κB(NFκB)及人核因子κB抑制蛋白α(IKβα)的表达水平。结果与阴性对照组相比,CSM+LPS组中IL-8、MDA含量及PDE-4活性显著性上升,抗氧化酶SOD、CAT活性、cAMP水平及PKA活性显著性下调,p65和IKβα的磷酸化水平增高。采用槲皮素预处理细胞后,与CSM+LPS组相比,槲皮素可显著性抑制IL-8释放,提升SOD及CAT酶活性并下调MDA的表达。此外,槲皮素可抑制PDE-4活性,上调cAMP及PKA水平,同时抑制p-p65及p-IKβα的表达。结论在CSM+LPS诱导的BEAS-2B细胞AECOPD模型中,槲皮素通过抑制PDE-4活性,激活cAMP/PKA信号通路,抑制NFκB的活化,抑制IL-8的释放。同时,槲皮素通过上调抗氧化酶活性发挥抗氧化应激作用。本研究结果揭示槲皮素具有预防AECOPD发生发展的潜在药理作用。OBJECTIVE To investigate the preventive effect of quercetin on acute exacerbation of chronic obstructive pulmonary disease(AECOPD) and its related mechanism, in order to provide theoretical basis for the clinical application of quercetin. METHODS Human bronchial epithelial BEAS-2 B cells were stimulated with cigarette smoke extract(CSM,4%,v/v) plus lipopolysaccharide(LPS,10 μg·mL-1) to establish AECOPD cell model.The cells were pretreated with quercetin(10 μmol·L-1) for 1 hour, and then stimulated with CSM plus LPS for 24 hours.Cells and supernatant in culture medium were collected.The content of interleukin-8(IL-8) in culture solution was detected by enzyme-linked immunosorbent assay(ELISA),the level of malondialdehyde(MDA) in cells was detected by thiobarbituric acid reaction method, and the levels of superoxide dismutase(SOD),catalase(CAT),phosphodiesterase-4(PDE-4),cyclic adenosine monophosphate(cAMP) and protein kinase A(PKA) in cells were detected by kit.The expressions of nuclear factor kappa B(NFκB) and human nuclear factor kappa B inhibitor protein α(IKβα) were detected by western blot. RESULTS Compared with the control group, the levels of IL-8,MDA and PDE-4 in CSM+LPS group were significantly increased, the activities of SOD,CAT cAMP and PKA were significantly decreased, and the phosphorylation levels of p65 and IKβα were increased.Compared with CSM+LPS group, quercetin significantly inhibited the release of IL-8,increased the activities of SOD and CAT,and down regulated the expression of MDA.In addition, quercetin could inhibit the activity of PDE-4,up-regulate the levels of cAMP and PKA,and inhibit the expressions of p-p65 and p-IKβα. CONCLUSION In the CSM+LPS-induced AECOPD model of BEAS-2 B cells, quercetin inhibits the release of IL-8 via activating PDE-4/cAMP/PKA and inhibiting NFκB signaling pathway.At the same time, quercetin exerts an anti-oxidative effect by up-regulating the activity of antioxidant enzymes.This results reveal that quercetin has potential pharmacological effects

关 键 词：槲皮素 急性加重慢性阻塞性肺疾病 氧化应激 环磷酸腺苷 核因子ΚB 

分 类 号：R969[医药卫生—药理学]