摘要
Objective This study aimed to analyze the mechanism of action of Huangqi(Astragalus Radix,HQ)-Jinyingzi(Rosae Laevigatae Fructus,JYZ)in the treatment of benign prostatic hyperplasia(BPH)based on network pharmacology and to verify the prediction through animal experimentation.Methods Based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN-TCM)databases,and literature,the active components and related target genes of HQ and JYZ were screened.The BPH target genes were screened based on the DisGeNET and GeneGards databases,and Excel was used to merge and remove duplicates.The Perl language was used to obtain drug-BPH target genes by intersecting shared target genes.A drug-component-target gene network diagram was constructed using Cytoscape software.The drug-BPH intersection target genes were inputted into the STRING database,and the key target genes were selected according to the degree algorithm.The output formed the basis for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses to determine the potential mechanism of HQ and JYZ in BPH treatment.High,medium,and low doses of HQ and JYZ extract were used to intervene in BPH rats,and then the prostate volume,wet weight,and prostate index of the BPH rats were determined.Changes in prostate histopathology and microvessel density(MVD)were evaluated using immunohistochemistry,and the optimal HQ and JYZ extract dose was confirmed.Finally,the optimal dose was used to intervene in a BPH rat model,and AKT1 and VEGF expressions were examined by immunohistochemistry.Results Based on network pharmacology,33 active components and 772 target genes were identified from HQ and JYZ,along with 817 BPH target genes and 112 drug-BPH common target genes.Among them were 10 key target genes,including AKT1,JUN,MAPK1,IL-6,TNF,ESR1,and VEGFA.KEGG enrichment analysis revealed 135 signaling pathways,including PI3K/AKT,I
目的基于网络药理学探讨黄芪-金樱子治疗良性前列腺增生症(BPH)的作用机制及作用靶点的实验验证。方法通过检索TCMSP、BATMAN-TCM数据库以及文献查阅,筛选出黄芪和金樱子的有效活性成分及相关靶基因。检索GeneCards、DisGeNET数据库,使用Excel软件合并去重,得到良性前列腺增生症相关靶基因。使用Perl语言获取“药物-疾病”交集靶基因数据库。使用Cytoscape 3.7.2软件构建“中药-成分-靶基因网络图”。将“药物-疾病”交集靶基因输入到STRING数据库,依据Degree算法筛选出核心靶基因。在此基础上,采用GO富集分析和KEGG通路富集探析黄芪-金樱子治疗BPH可能的作用机制和通路。采用黄芪-金樱子提取液低、中、高剂量干预BPH大鼠模型,检测大鼠前列腺体积、湿重及前列腺指数,免疫组织化学法观测前列腺组织病理形态学及微血管密度(MVD)改变,确定最佳治疗剂量。在此基础上,以最佳剂量干预BPH大鼠模型,免疫组织化学法检测AKT1和VEGF的表达。结果通过网络药理学,共筛选出黄芪和金樱子活性成分33个,相关靶基因772个,良性前列腺增生症靶基因817个,药物-疾病共同靶点112个,其中核心靶点10个,AKT1、JUN、MAPK1、IL-6、TNF、ESR1、VEGFA等基因排名靠前。KEGG富集分析得到135条通路,主要包括PI3K/AKT、IL-17、TNF、p53、MAPK、VEGF、JAK-STAT、NF-κB通路等。动物实验表明,黄芪和金樱子能显著改善BPH大鼠前列腺体积、湿重、前列腺指数及前列腺组织病理形态,降低MVD,还可抑制大鼠前列腺组织中AKT1和VEGF的表达,促进上皮细胞的凋亡,抑制血管新生,进一步验证了网络药理学预测中药作用于BPH多靶点,多途径的可能性。结论黄芪和金樱子可通过多靶点、多途径协同治疗BPH,其可能的作用机制为通过调节AKT1、VEGF蛋白及与细胞凋亡、血管新生、炎症相关的PI3K/AKT、VEGF信号通路来治疗BPH,具有潜在�
基金
We thank for the funding support from the Hunan Provincial Science and Technology Department(No.2020JJ4068 and No.2018SK4012).
