摘要
目的基于网络药理学联合分子对接技术探讨肥大细胞(Mast cells,MCs)脱颗粒抑制剂—色苷酸钠(Sodium Cromolyn,SCG)预防低氧性急性肺动脉高压(Acute Pulmonary Hypertension,APH)的作用机制。方法(1)从Swiss Target Predicition、TargetNet、UniProt、OMIM、GeneCards、NCBI gene数据库中获取色苷酸(Cromolyn)治疗APH的靶基因,使用Venny 2.1.0在线工具绘制Venn图并获取交集基因。通过String数据库构建蛋白质相互作用(protein-protein interaction,PPI)网络。使用Cytoscape软件对PPI网络进行拓扑分析,使用DAVID数据库做基因本体(Gene Ontology,GO)功能分析、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,并筛选出相关信号通路,使用Autodock Vina软件进行分子对接分析。(2)给予SD大鼠SCG预处理(100 mg/kg,ig,Bid,3 d),利用低压舱(模拟海拔7000 m,12 h)构建急性低氧动物模型。将SD大鼠随机分为低氧对照组(HC组,12只)和低氧SCG处理组(HSCG组,12只)。将低压舱海拔由7000 m降至3500 m,应用颈外静脉插管术监测肺动脉压(Pulmonary arterial pressure,PAP),通过蛋白质印迹(Western Blot,WB)实验检测FcεRⅠ、SYK、mTOR蛋白和p-mTOR(Ser2448)蛋白在SD大鼠肺组织中的表达水平。结果(1)基于网络药理学分析方法筛选出FcεRⅠ蛋白和mTOR信号通路及核心蛋白FcεRⅠ、SYK、mTOR。FcεRⅠ、SYK、mTOR分别与Cromolyn对接,均具有良好的结合活性,构象较稳定。(2)SD大鼠mPAP结果:与HC组比较,HSCG组的mPAP下降(P<0.05)。(3)肺组织WB结果:与HC组比较,HSCG组SD大鼠肺组织中的FcεRⅠ、SYK、mTOR蛋白和p-mTOR(Ser2448)蛋白表达均下降(P<0.05),p-mTOR(Ser2448)/mTOR亦下降(P<0.05)。结论经SCG预处理,可减弱急性低氧应激SD大鼠肺组织MCs中的FcεRⅠ、SYK、mTOR蛋白和p-mTOR(Ser2448)蛋白表达、降低p-mTOR(Ser2448)/mTOR,致FcεRⅠ、mTOR信号通路的激活水平降低,进而抑制肺组织中MCs的活化脱颗粒过程,减少收�
Objective To explore the action mechanism of mast cells(MCs)degranulation inhibitor sodium cromolyn(SCG)in preventing acute pulmonary hypertension(APH)combined network pharmacology with molecular docking technology.Methods(1)The target genes of cromolyn for APH were obtained from Swiss Target,TargetNet,UniProt,OMIM,GeneCards and NCBI gene databases.Venny 2.1 online tool was adopted to draw the Venn diagram and obtain intersection genes.A protein-protein interaction(PPI)network was constructed through String database.Topological analysis of PPI network was carried out by using software Cytoscape.Gene Ontology(GO)function analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis was carried out by using DAVID database,and relevant signal pathways were screened out.Molecular docking analysis was carried out by using software Autodock Vina.(2)SD rats were given SCG(100 mg/kg,ig,Bid,3 d)pre-treatment.The acute hypoxia animal model was replicated by using hypobaric chamber(simulated altitude of 7,000 m,12 h).SD rats were randomly divided into hypoxia control group(HC group,12 rats)and hypoxia SCG treatment group(HSCG group,12 rats).The elevation of hypoxic chamber was lowered from 7,000 m to 3,500 m.The pulmonary arterial pressure(PAP)was monitored by external carotid vein intubation.The expression levels of FcεRⅠ,SYK,mTOR,p-mTOR(ser 2448)in lung tissue of SD rats were detected by Western Blot(WB)assay.Results(1)FcεR I and mTOR signaling pathways as well as core proteins including FcεR I,SYK and mTOR were selected on the basis of network pharmacological analysis.FcεR I,SYK and mTOR were docked with cromolyn respectively,and all showed good binding activity and stable conformation.(2)mPAP results of SD rats:mPAP decreased in HSCG group compared with HC group(P<0.05).(3)WB results of lung tissue:Compared with HC group,the protein expression of FcεRⅠ,SYK,mTOR and p-mTOR(ser2448)in lung tissue of SD rats in HSCG group decreased(P<0.05),and p-mTOR(ser2448)/mTOR also declined(P<0.05).Conclusion A
作者
史玉玲
张长荣
范勇
李海霞
杨国娟
马爽
刘杰
SHI Yuling;ZHANG Changrong;FAN Yong;LI Haixia;YANG Guojuan;MA Shuang;LIU Jie(Medical Department of Qinghai University,Xining 810001,China;Qinghai University Affiliated Hospital,Xining 810001,China)
出处
《中国高原医学与生物学杂志》
CAS
2024年第3期156-166,共11页
Journal of Chinese High Altitude Medicine & Biology
基金
青海省科技厅应用基础研究项目(2020-ZJ-772)。
关键词
色苷酸钠
急性肺动脉高压
低氧
肥大细胞
网络药理学
分子对接
sodium cromolyn(SCG)
acute pulmonary hypertension(APH)
hypoxia
mast cells(MCs)
network pharmacology
moleculardocking
