青藤碱激活Keap1/Nrf2信号通路抑制氧化应激和肺纤维化  

Sinomenine inhibits oxidative stress and pulmonary fibrosis by activating the Keap1/Nrf2 signaling pathway

作　　者：刘理静 钱红 孟庆欣 张翔 贺彬 贺兼斌 魏英民 LIU Lijing;QIAN Hong;MENG Qingxin;ZHANG Xiang;HE Bin;HE Jianbin;WEI Yingmin(School of Medicine,Changsha Social Work College,Changsha 410004,Hunan,China;School of Nursing,Hunan University of Medicine,Huaihua 418000,Hunan,China;Department of Respiratory and Critical Care Medicine,Huaihua Hospital Affiliated to University of South China,First People's Hospital of Huaihua,Huaihua 418000,Hunan,China)

机构地区：[1]长沙民政职业技术学院医学院,长沙410004 [2]湖南医药学院护理学院,怀化418000 [3]南华大学附属怀化医院,怀化市第一人民医院呼吸与危重症医学科,怀化418000

出　　处：《中国临床药理学与治疗学》2023年第9期979-987,共9页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：湖南省教育厅资助科研项目(21C1593);湖南省自然科学基金资助项目(2018JJ2279);长沙民政职业技术学院教授、博士科研项目(2023JB28)。

摘　　要：目的:探讨青藤碱(SIN)对氧化应激损伤、肺纤维化的保护作用及与Kelch样ECH相关蛋白1(Keap1)/核因子E_(2)相关因子2(Nrf2)信号通路的关系。方法:通过过氧化氢(H_(2)O_(2))诱导MRC-5细胞建立氧化应激损伤模型,并予SIN处理,采用CCK-8法检测细胞活力,生化试剂盒检测丙二醛(MDA)含量及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性,Western blot检测Keap1、Nrf2蛋白表达。将30只SD大鼠随机分为对照组、博莱霉素A5(BLM)组和BLM+SIN组,每组包含10只动物,BLM组、BLM+SIN组气管内注射BLM建立大鼠肺纤维化模型,对照组气管内注射等体积9 g/L氯化钠溶液,造模后次日,BLM+SIN组以SIN灌胃,其余两组给予9 g/L氯化钠溶液灌胃,第28天处死大鼠,分离肺组织,通过HE和Masson染色观察肺组织病理学改变,测定肺组织MDA含量及SOD、GSH-Px、CAT活性,Western blot检测Keap1、Nrf2蛋白表达。结果:相对于H_(2)O_(2)组,SIN干预后MRC-5细胞活力增加,MDA含量降低,SOD、GSH-Px和CAT活性升高,下调Keap1表达,促进Nrf2核移位。与BLM组相比,SIN给药降低大鼠肺泡炎、肺纤维化病变和评分及肺组织MDA含量,增加肺组织SOD、GSH-Px和CAT活性,下调肺组织Keap1表达,升高胞核Nrf2水平。结论:SIN可能通过激活Keap1/Nrf2信号通路抑制氧化应激损伤,减轻大鼠肺纤维化。AIM:To explore the protective effects of sinomenine(SIN)on oxidative stress and pulmonary fibrosis and its relationship with the Kelch-like ECH-associated protein 1(Keap1)/nuclear factor erythroid 2-related factor 2(Nrf2)signaling pathway.METHODS:MRC-5 cells were treated with hydrogen peroxide(H_(2)O_(2))to establish the oxidative stress injury model,followed by administration with SIN.Cell viability was detected using the CCK-8 method.The biochemical kits were employed to measure malondialdehyde(MDA)content and superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and catalase(CAT)activities.The protein expression of Keap1 and Nrf2 was examined by western blot.Thirty SD rats were randomly divided into control group,bleomycin A5(BLM)group and BLM+SIN group,with 10 animals in each group.Bleomycin A5 were intratracheally administered to the rats in BLM group and BLM+SIN group to establish the pulmonary fibrosis model.The rats in control group received the same volume of 9 g/L sodium chloride solution.The second day after model construction,the rats in BLM+SIN group were gavaged with SIN,while the rats in the other two groups were treated with 9 g/L sodium chloride solution.On day 28,all rats were sacrificed.Pulmonary tissue was isolated,and HE and Masson staining was performed to observe the pathological changes.The MDA content and SOD,GSH-Px and CAT activities in pulmonary tissue were evaluated.Western blot was used to assay pulmonary tissues Keap1 and Nrf2 protein expression.RESULTS:When compared with H_(2)O_(2) group,SIN treatment increased cell viability,decreased MDA content,elevated SOD,GSH-Px and CAT activities,down-regulated Keap1 expression,and promoted nuclear translocation of Nrf2 in MRC-5 cells.In comparison with BLM group,administration of SIN decreased alveolitis and pulmonary fibrosis pathological changes and scores as well as pulmonary tissue MDA content,enhanced pulmonary tissues SOD,GSH-Px and CAT activities,down-regulated pulmonary tissues Keap1 expression,and raised Nrf2 levels in the nucleus.

关 键 词：青藤碱 肺纤维化 氧化应激 Kelch样ECH相关蛋白1 核因子E2相关因子2 

分 类 号：R285.5[医药卫生—中药学] R392.12[医药卫生—中医学] R563