香青兰提取物对博来霉素所致肺纤维化大鼠的干预作用及机制研究  

Intervention effect and mechanism of Dracocephalum moldavica L.extract on bleomycin-induced pulmonary fibrosis in rats

作　　者：孙小玉 陈丽 高瑞芳 于慧 靳敏[1] SUN Xiaoyu;CHEN Li;GAO Ruifang;YU Hui;JIN Min(Inner Mongolia University of Science and Technology Baotou Medical Colleague a.School of Public Health;Colleague of Basic Medicine and Forensic Medicine,Inner Mongolia Autonomous Region,Baotou,014000,China;Laboratory,Yuhua District Center for Disease Control and Prevention,Shijiazhuang,Hebei 050023,China)

机构地区：[1]内蒙古科技大学包头医学院公共卫生学院,内蒙古包头014000 [2]内蒙古科技大学包头医学院基础医学与法医学院,内蒙古包头014000 [3]石家庄市裕华区疾病预防控制中心检验科,河北石家庄050023

出　　处：《环境与职业医学》2024年第12期1384-1392,共9页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金地区项目(82260092);2022年度包头医学院创新团队发展计划项目(bycxtd-11)。

摘　　要：[背景]环境污染和特定职业暴露加剧了肺纤维化问题,肺纤维化发病机制复杂且缺乏有效治疗药物。香青兰提取物可通过抗炎抗焦亡途径缓解肺纤维化,但其对肺纤维化的防治机制尚不明确。[目的]结合网络药理学和动物实验分析香青兰提取物抗肺纤维化的作用靶点及潜在机制。[方法]借助中国知网、中药系统药理学数据库与分析平台(TCMSP)检索香青兰提取物的活性成分,利用Gene Cards、DisGeNET获取肺纤维化相关疾病靶点,采用相互作用基因库检索工具(STRING)数据库和Cytoscape软件构建蛋白相互作用网络图(PPI),将预测的潜在靶点通过注释、可视化、集成发现数据库(DAVID)进行基因本体论(GO)及京都基因与基因组百科全书(KEGG)通路富集分析,并采用分子对接进行验证。将32只大鼠随机分为对照组、模型组、香青兰提取物低剂量组(100 mg·kg^(-1))、香青兰提取物高剂量组(400 mg·kg^(-1)),每组8只。除对照组外,采用博来霉素(5 mg·kg^(-1))气管滴注法构建肺纤维化大鼠模型,对照组滴注等量的生理盐水。建模后,香青兰提取物剂量组大鼠灌胃相应剂量的药物,对照组和模型组分别灌胃给予等体积生理盐水,每天1次。连续28 d后处死,收集肺组织。通过病理切片染色观察大鼠肺组织病理学变化,免疫印迹法(WB)检测肺组织中与纤维化相关蛋白I型胶原蛋白(Col-I)、α-平滑肌肌动蛋白(α-SMA)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)蛋白表达水平,实时荧光定量PCR(RT-qPCR)检测肺组织中α-SMA、Col-I mRNA水平,酶联免疫吸附试验(ELISA)检测大鼠肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1β(IL^(-1)β)水平。[结果]通过数据库筛选共获得香青兰提取物378个有效成分,肺纤维化相关靶点1 611个;筛选获得香青兰提取物抗肺纤维化潜在靶点574个。以Degree值为标准筛选出关键靶点为白蛋白(ALB)、肿[Background]Exposures to environmental pollution and specific occupational hazards exacerbate pulmonary fibrosis which has a complex pathogenesis and lacks effective therapeutic drugs.The extract from Dracocephalum moldavica L.can alleviate pulmonary fibrosis through anti-inflammatory and anti-pyroptosis pathways,but its mechanism of prevention and treatment for pulmonary fibrosis remains unclear.[Objective]To elucidate the targets and potential mechanism underlying the anti-pulmonary fibrosis efficacy of Dracocephalum moldavica L.extract by employing an amalgamation of network pharmacology and empirical verification.[Methods]The chemical composition of the extract of Dracocephalum moldavica L.was retrieved with the help of China National Knowledge Infrastructure(CNKI)and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).The disease targets related to pulmonary fibrosis were inquired using Gene Cards and DisGeNET.A protein-protein interaction(PPI)was constructed using the Search Tool for the Retrieval of Interacting Genes(STRING)database and Cytoscape software.The predicted potential targets were analyzed by the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses through the Database for Annotation,Visualization and Integrated Discovery(DAVID)and validated by molecular docking.Thirty-two rats were randomly divided into a control group,a model group,a low-dose group of Dracocephalum moldavica L.extract(100 mg·kg^(-1)),and a highdose group of Dracocephalum moldavica L.extract(400 mg·kg^(-1)),with eight rats in each group.A rat model of pulmonary fibrosis was constructed using bleomycin(5 mg·kg^(-1))intratracheal instillation,and an equal volume of saline was instilled into the control group.After modelling,400 and 100 mg·kg^(-1) of Dracocephalum moldavica L.extract were given the high-dose and low-dose groups by gavage,and an equal volume of saline was given by gavage to the control group and the model group,once per day,for consecu

关 键 词：肺纤维化 香青兰提取物 磷脂酰肌醇3-激酶/蛋白激酶B 炎症 网络药理学 动物实验验证 

分 类 号：R13[医药卫生—劳动卫生]