基于网络药理学探讨构树治疗特异性皮炎的作用机制

Mechanism of Broussonetia papyriferain Treating Atopic Dermatitis Based on Network Pharmacology

摘要

目的 通过网络药理学、分子对接技术及实验验证，探讨构树（Broussonetia papyrifera, BP）治疗特异性皮炎（atopic dermatitis，AD）的作用靶点及机制。方法 建立 2，4-二硝基氯苯（2，4-dichloronitrobenzene，DNCB）诱导的小鼠 AD 模型，分为正常对照组（Nor）、模型组（Mod）、地塞米松组（DEX）、构树提取物低剂量组（L-BP）和高剂量组（H-BP），连续给药 12 天。对 AD 小鼠进行皮损情况观察、皮炎评分及病理染色，确定 BP 治疗 AD 的药效。利用 HERB、TCMSP 和 SwissTargetPrediction 数据库预测 BP 的活性成分及其作用靶点，并结合 GeneCards、DisGeNET 和 OMIM 数据库筛选出与 AD 相关的靶点。通过 Venny 制作韦恩图，筛选出 BP 与 AD 的共同靶点。利用 STRING 和 Cytoscape 软件进行蛋白质相互作用（protein-protein interaction，PPI）网络分析，以 degree 值排前 10 的靶点作为核心靶点。通过基因本体论（Gene Ontology，GO）功能和京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）通路富集分析 BP 治疗 AD 的可能作用机制，并使用 AutoDock 软件评估 AD 的活性成分和核心靶点之间的结合亲和力。最后，以 Western blot 实验等验证网络药理学预测的结果。结果 体内实验结果显示，与模型组相比，BP 能显著提高小鼠的表皮厚度，降低皮炎评分，改善 AD 症状。经网络药理学分析，从 BP 的 115 个靶点和 AD 的 1 815 个靶点中筛选出 47 个交集靶点。PPI 网络拓扑分析确定了 10 个核心靶点，包括ESR1、AKT1、PTGS2、HIF1A、NF-κB1等。分子对接结果显示，BP 活性成分与这些核心靶点结合能力较好。GO 功能富集结果显示，有 74 个分子功能、22 个细胞成分、167 个生物过程。KEGG 通路富集分析显示，PI3K-AKT、NF-κB 信号通路可能是 BP 治疗 AD 的重要途径。验证结果显示，BP 可通过降低 PI3K、IKB、p65、AKT 的磷酸化水平，抑制 PI3K/AKT 和 NF-κB 信号通路的激活。结论 BP 可通过调节 PI3K/AKT 和 NF-κB 信号通路发挥治疗 AD 的作用，为 BP 治疗 AD 的后续深入研究和潜在临床应用提供参考。

Abstract

Objective This study aimed to explore the therapeutic targets and mechanisms of Broussonetia papyrifera(BP) in the treatment of atopic dermatitis (AD) through network pharmacology, molecular docking, and animal experiments. Methods An AD mouse model was established through 2,4-dinitrochlorobenzene (DNCB) induction. The mice were divided into five groups: normal control group (Nor), model group (Mod), dexamethasone group (DEX), low-dose BP extract group (L-BP), and high-dose BP extract group (H-BP), and treated for 12 consecutive days. The therapeutic effects of BP on AD were evaluated by observing the skin damage, scoring the dermatitis, and conducting histopathological staining. The active components of BP and their targets were predicted using the HERB, TCMSP, and SwissTargetPrediction databases. Targets related to AD were screened using the GeneCards, DisGeNET, and OMIM databases. The common targets of BP and AD were identified by Venny and visualized using a Venn diagram. Protein-protein interaction (PPI) network analysis was performed using STRING and Cytoscape software, and the top 10 targets with the highest degree values were identified as core targets. The possible mechanism of BP in treating AD was analyzed using Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. The binding affinity between the active components of BP and the core targets was assessed using AutoDock software. Finally, Western blot experiments were conducted to verify the predictions made by network pharmacology. Results The in vivoexperimental results showed that, compared with the model group, BP significantly increased the epidermal thickness of DNCB-induced AD mice, reduced the dermatitis score, and improved AD symptoms. Network pharmacology analysis identified 47 common targets from 115 BP targets and 1 815 AD targets. PPI network topology analysis determined 10 core targets, includingESR1, AKT1, PTGS2, HIF1A, and NF-κB1. Molecular docking results indicated that the active components of BP had good binding affinity with these core targets. GO function enrichment analysis revealed 74 molecular functions, 22 cellular components, and 167 biological processes. KEGG pathway enrichment analysis suggested that the PI3K-AKT and the NF-κB signaling pathway might be important pathways for BP in treating AD. Animal experiments demonstrated that BP could inhibit the activation of the PI3K/AKT and NF-κB signaling pathways by reducing the phosphorylation levels of PI3K, IKB, p65, and AKT. Conclusion BP exerts its therapeutic effects on AD by modulating the PI3K/AKT and NF-κB signaling pathways, which provides a scientific basis for further in-depth research and potential clinical application of BP in the treatment of AD.

关键词

网络药理学 / 分子对接 / 特异性皮炎 / 构树 / PI3K-AKT 信号通路 / 作用机制

Key words

network pharmacology / molecular docking / atopic dermatitis / Broussonetia papyrifera / PI3K-AKT signaling pathway / mechanism

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基金资助

国家级大学生创新创业训练计划项目(202310573012)