基于网络药理学和分子对接技术分析芡实-莲子抗乳腺癌的作用机制

Analysis of the Anti-breast Cancer Mechanism of Euryales Semen-Nelumbinis Semen Based on Network Pharmacology and Molecular Docking Technology

摘要:

目的：应用网络药理学与分子对接技术分析芡实-莲子抗乳腺癌的潜在作用机制。方法：从 TCMSP、UniProt等数据库获取芡实、莲子的活性成分及靶点，从GeneCards和OMIM数据库获取乳腺癌相关靶 点，利用Venny 2.1.0获取共同靶点。通过DAVID数据库进行基因本体（GO） 功能和京都基因与基因组百科全 书（KEGG） 通路富集分析，利用STRING和Cytoscape构建蛋白互作（PPI） 网络及“药物-活性成分-疾病靶 点”网络，使用Autodock和PyMOL进行分子对接验证。利用TIMER2.0分析核心靶点基因表达与免疫细胞浸润 的相关性及其预后。结果：筛选出芡实、莲子中5个活性成分为β-胡萝卜素、维生素E、N-去甲荷叶碱、阿诺 新碱、（+） -阿美帕维碱，疾病靶点2 510个，芡实-莲子抗乳腺癌的潜在作用靶点101个。筛选出核心靶点为 雌激素受体1 （ESR1）、雌激素受体2 （ESR2）、雄激素受体（AR）、Rho 相关卷曲螺旋形成蛋白激酶 2（ROCK2）、磷脂酰肌醇-4，5-二磷酸3-激酶催化亚基γ型（PIK3CG）、细胞色素P450 1A2（CYP1A2）、前列 腺素内过氧化物合酶2（PTGS2）、丝裂原活化蛋白激酶8（MAPK8）。GO和KEGG分析表明潜在治疗靶点显著 富集于细胞增殖、凋亡、血管生成及免疫反应等通路。分子对接证实主要活性成分与核心靶点结合良好。免疫 浸润分析表明，芡实-莲子共同靶向的ESR2、PIK3CG和PTGS2低表达以及ESR1高表达均可减少CD8+ T细胞浸 润，缩短生存期；AR和ROCK2低表达以及CYP1A2高表达均可增加CD8+ T细胞浸润，延长生存期。AR高表 达减少静息肥大细胞浸润，延长生存期。且CYP1A2 和MAPK8 高表达均增加CD4+ T 细胞浸润；CYP1A2 和 PTGS2高表达均减少调节性T细胞（Tregs） 浸润。结论：芡实-莲子可能通过多靶点调控肿瘤细胞生物学行为， 重塑肿瘤免疫微环境发挥抗乳腺癌作用。

Abstract:

Abstract： Objective： To analyze the potential anti-breast cancer mechanism of Euryales Semen-Nelumbinis Semen using network pharmacology and molecular docking techniques. Methods：Active ingredients and corresponding targets of Euryales Semen and Nelumbinis Semen were obtained from databases such as Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and Universal Protein Resource （UniProt）. Breast cancer-related targets were retrieved from the Human Gene Database （GeneCards） and Online Mendelian Inheritance in Man （OMIM） databases. Common targets were identified using Venny 2.1.0. Gene Ontology （GO） function and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were performed using the Database for Annotation， Visualization， and Integrated Discovery （DAVID）. A protein-protein interaction （PPI） network and a "drug-active ingredient-disease target" network were constructed using STRING and Cytoscape. Molecular docking validation was conducted using AutoDock and PyMOL. TIMER2.0 was employed to analyze correlations between the expression of core targets， immune cell infiltration， and prognosis. Results： Five active ingredients from Euryales Semen and Nelumbinis Semen were identified： β-carotene， vitamin E， N-Nornuciferine， anonaine， and （+）- amepavine. Among 2 510 breast cancer-related targets， 101 potential targets for the anti-breast cancer effect of Euryales Semen-Nelumbinis Semen were obtained. Key targets were identified as estrogen receptor 1（ ESR1），estrogen receptor 2 （ESR2）， androgen receptor （AR）， Rho-associated coiled-coil containing protein kinase 2 （ROCK2）， phosphoinositide-3-kinase catalytic subunit gamma （PIK3CG）， cytochrome P450 1A2 （CYP1A2）， prostaglandinendoperoxide synthase 2 （PTGS2）， and mitogen-activated protein kinase 8 （MAPK8）. GO and KEGG analyses indicated that the targets were significantly enriched in pathways related to cell proliferation，apoptosis，angiogenesis， and immune responses. Molecular docking confirmed strong binding affinity between the main active ingredients and core targets. Immune infiltration analysis revealed that low expression of ESR2， PIK3CG， and PTGS2， as well as high expression of ESR1，all commonly targeted by Euryales Semen and Nelumbinis Semen，were associated with reduced CD8+ T cell infiltration and shorter survival. In contrast，low expression of AR and ROCK2，along with high expression of CYP1A2，were associated with increased CD8+ T cell infiltration and prolonged survival. Moreover，high expression of AR was associated with reduced resting mast cell infiltration，which was linked to longer survival. Furthermore，high expression of CYP1A2 and MAPK8 both correlated with increased CD4+ T cell infiltration；whereas high expression of CYP1A2 and PTGS2 both correlated with reduced regulatory T cell （Tregs） infiltration. Conclusion：Euryales Semen and Nelumbinis Semen may exert anti-breast cancer effects by regulating tumor cell biological behaviors through multiple targets and remodeling the tumor immune microenvironment.