基于生物信息学的杆努尽烟治疗慢性支气管炎大鼠的分子机制  

Mechanisms of Lysionotus pauciflorus Maxim.for the treatment of chronic bronchitis based on bioinformatics

作　　者：胡子怡 冯静 杨向前 王茜 赵新越 吴宁 钟曦 吴昌学[2] HU Ziyi;FENG Jing;YANG Xiangqian;WANG Qian;ZHAO Xinyue;WU Ning;ZHONG Xi;WU Changxue(Chemical and Biochemical Laboratory,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;Key Laboratory of Medical Biology,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院化学与生物化学实验室,贵州贵阳550025 [2]贵州医科大学医学分子生物学重点实验室,贵州贵阳550004

出　　处：《贵州医科大学学报》2024年第4期489-500,共12页Journal of Guizhou Medical University

基　　金：国家自然科学基金(8206776);贵州省科技计划项目(黔科合LH字〔2020〕1Y388);贵州省大学生创新创业计划项目(202010660005,S202210660094,S202310660047)。

摘　　要：目的基于生物信息学探讨杆努尽烟治疗慢性支气管炎(CB)大鼠的分子机制。方法采用Linpinski规则筛选出杆努尽烟活性成分,UniProt预测潜在靶点,取Gene Cards与疾病靶点交集,String创建蛋白互作网络并创建“成分-靶点-通路”网络,基因本体数据库(GO)和京都基因与基因组百科全书(KEGG)富集分析交集靶点,AutoDock模拟活性成分-核心靶点的对接模式;30只大鼠随机均分为空白组(0.9%NaCl)、模型组(0.9%NaCl)、阳性药物(1g/kg桂龙咳喘宁)组及低剂量(2 g/kg)、高剂量(8 g/kg)杆努尽烟组,除空白组外其余组大鼠进行CB造模,造模结束后麻醉处死各组大鼠1只,取肺组织制作切片采用苏木精-伊红染色法(HE)判断造模成功与否;再按上述各组处理灌胃给药,1次/d、持续28 d,干预结束后麻醉处死余下各组大鼠,取肺组织和心脏取血,采用Western blot和反转录-聚合酶链反应(RT-PCR)检测肺组织中蛋白激酶B(AKT1)、过氧化物酶体增殖物激活受体γ(PPARγ)及激活核因子-κB p65(NF-κB p65)蛋白和mRNA表达,采用酶联免疫吸附测定(ELISA)法检测各组大鼠血清肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)及干扰素γ(INF-γ)水平。结果杆努尽烟活性成分18个交集靶点92个,核心靶点AKT1、PPARγ等,GO-KEGG富集主要涉及免疫和炎症相关的磷脂酰肌醇3-酶d/蛋白激酶B(PI3K/Akt)、丝裂原活化蛋白激酶(MAPK)信号通路,分子对接示杆努尽烟活性成分与AKT1、PPARγ结合稳定;与空白组比较,模型组大鼠肺组织PPARγ蛋白及mRNA表达下降,NF-κB p65、AKT1蛋白及mRNA表达上升(P<0.05);与模型组比较,阳性药物组、低剂量杆努尽烟组、高剂量杆努尽烟组大鼠肺组织PPARγ蛋白及mRNA表达量均有不同程度的上升,NF-κB p65、AKT1蛋白及mRNA表达量均有一定程度的下降,高剂量杆努尽烟组大鼠肺组织表达差异最为明显(P<0.05);模型组和低剂量杆努尽烟组大鼠血清TNF-α、IL-1β及IObjective To explore the molecular mechanisms of Lysionotus pauciflorus Maxim.in treating chronic bronchitis(CB)in rats based on bioinformatics.Methods Active components of Lysionotus pauciflorus Maxim.were screened using the Lipinski rule.Potential targets were predicted with UniProt,intersected with disease targets from Gene Cards,and a protein interaction network was created with string to establish a"Component-Target-Pathway"network.Intersection targets were enriched and analyzed using the Gene Ontology Database and the Kyoto Encyclopedia of Genes and Genomes(GO-KEGG),and the docking mode of active components with core targets was simulated with AutoDock.Thirty rats were randomly divided into five groups:control(0.9%NaCl),model(0.9%NaCl),positive drug(1g/kg Guilong cough and asthma relief),low-dose(2 g/kg),and high-dose(8 g/kg)Lysionotus pauciflorus Maxim.groups.After CB modeling,lung tissues were collected for HE staining to judge the success of the modeling;gastric administration was then performed once a day for 28 days.Western blot and RT-PCR were used to detect the expression of proteins and mRNAs such as protein kinase B(AKT1),peroxisome proliferator-activated receptor γ(PPARγ),and nuclear factor-κB(NF-κB p65)in lung tissues;ELISA was used to detect serum levels of tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),and interferonγ(INF-γ).Results Lysionotus pauciflorus Maxim.had 18 active components with 92 intersecting targets,including core targets like AKT1 and PPARγ.GO-KEGG enrichment mainly involved immune and inflammation-related phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt),mitogen-activated protein kinase(MAPK)signaling pathways.Compared with blank group,PPARγprotein and mRNA expression in lung tissue of model group decreased,while NF-κB p65 and AKT1 protein and mRNA expression increased(P<0.05).Compared with model group,PPARγprotein and mRNA expression in lung tissue of rats in positive drug group,low-Lysionotus pauciflorus group and high-Lysionotus pauciflorus group we

关 键 词：杆努尽烟 慢性支气管炎 生信分析 体内实验 肺组织 PI3K-AKT信号通路 

分 类 号：R562.2[医药卫生—呼吸系统]