摘要
目的利用网络药理学和细胞实验研究党参(Codonopsis Radix,CR)治疗肠黏膜炎(intestinal mucositis,IM)的潜在作用机制。方法从中药系统药理学数据库(TCMSP)中获取党参的化学成分,UniProt、SwissTargetPrediction数据库获得党参化学成分对应的靶点,在线人类孟德尔遗传数据库(OMIM)、GeneCards数据库获得疾病相关基因,微生信数据分析平台筛选党参和IM的交集靶点,获得党参治疗IM的作用靶点。使用Cytoscape软件绘制化学成分-靶点-疾病交互网络,利用STRING数据库获取蛋白相互作用(PPI)网络图,利用注释、可视化和综合发现数据库(DAVID)进行基因本体(GO)和京都基因和基因组百科全书(KEGG)通路富集分析。使用不同浓度的党参醇提物(0,50,100μg/mL)处理五氟尿嘧啶(5-FU,10μg/mL)刺激的小肠上皮细胞IEC-6,酶联免疫吸附实验(ELISA)检测细胞炎症因子水平,免疫组化检测细胞磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(AKT)蛋白表达,荧光法检测细胞内活性氧(ROS)含量。结果共筛选出党参活性成分21个,对应401个靶点。肠黏膜炎相关靶点2210个,交集靶点197个。GO和KEGG富集分析显示,党参治疗肠黏膜炎主要涉及PI3K/AKT信号通路等。细胞实验结果显示,100μg/mL党参醇提物显著降低细胞炎症因子肿瘤坏死因子-α(TNF-α)和白介素-6(IL-6)的含量(P<0.05),上调IL-4和IL-10的含量(P<0.01),而50μg/mL党参醇提物降低细胞炎症因子TNF-α和IL-6的效果不显著,降低IL-4和IL-10的含量(P<0.05);100μg/mL党参醇提物明显降低细胞ROS水平(P<0.01),下调PI3K和AKT的表达量(P<0.01),50μg/mL党参醇提物降低细胞AKT和PI3K水平(P<0.05),降低ROS的效果不显著。结论党参可能通过调控PI3K/AKT信号通路发挥治疗肠黏膜炎的作用。
Objective To investigate the potential mechanism of Codonopsis Radix(CR)in the treatment of intestinal mucositis(IM)using network pharmacology and verify it in cell experiments.Methods The chemical constituents of Codonopsis Radix were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database,the targets corresponding to the chemical components of Codonopsis Radix were obtained from UniProt and SwissTargetPrediction database,the disease-related genes were obtained from online Mendelian Inheritance in Man(OMIM)and GeneCards database,and the Codonopsis Radix and IM intersection targets were screened by bioinformatics data analysis platform,and the action targets of Codonopsis Radix for IM were obtained.Cytoscape software was used to construct the chemical compositional-target-disease interaction network,the STRING database was used to obtain PPI network,and the Database for Annotation,Visualization and Integrated Discovery(DAVID)database was used for gene ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Small intestinal epithelial cells IEC-6 stimulated by 5-fluorouracil(5-FU,10μg/mL)were treated with different concentrations of Codonopsis Radix alcohol extract(0,50,100μg/mL).Inflammatory cytokines were detected by enzyme-linked immunosorbent assay(ELISA),the expressions of PI3K and AKT were detected by immunohistochemistry,and the reactive oxygen species(ROS)content was detected by fluorescence method.Results A total of 21 active ingredients of Codonopsis Radix were selected,corresponding to 401 targets.There were 2210 targets related to intestinal mucositis and 197 targets of Codonopsis Radix-intestinal mucositis intersection.GO and KEGG enrichment analysis showed that Codonopsis Radix treated intestinal mucositis mainly via PI3K/AKT signaling pathway,etc.The alcohol extract of Codonopsis Radix(100μg/mL)decreased the contents of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)(P<0.05),and upregulated the contents of IL-4 and IL-10(P<0.01).The alcohol extract of Codonopsis Radix(50μg/mL)had no significant effect on the reduction of inflammatory factors(TNF-αand IL-6),and reduced the contents of IL-4 and IL-10(P<0.05).The alcohol extract of Codonopsis Radix(100μg/mL)decreased the level of ROS(P<0.01),and downregulated the expressions of PI3K and AKT(P<0.01).The alcohol extract of Codonopsis Radix(50μg/mL)decreased the levels of AKT and PI3K(P<0.05),but had no significant effect on ROS.Conclusion Codonopsis Radix may play a role in the treatment of intestinal mucositis by regulating PI3K/AKT signaling pathway.
作者
李德运
闫丽秋
王嘉静
周江韬
LI Deyun;YAN Liqiu;WANG Jiajing;ZHOU Jiangtao(Department of Traditional Chinese Medicine,School of Pharmacy,Shanxi Medical University,Taiyuan 030001,China;Department of Medicinal Chemistry,College of Pharmacy,Shanxi Medical University)
出处
《山西医科大学学报》
CAS
2024年第5期580-588,共9页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(81904031)。
关键词
党参
肠黏膜炎
炎症
网络药理学
PI3K/AKT信号通路
Codonopsis Radix
intestinal mucositis
inflammation
network pharmacology
PI3K/AKT signaling pathway
