目的 基于系统药理学和转录组学技术结合分子对接及实验验证，探讨金振口服液（Jinzhen Oral Liquid，JZ）改善下呼吸道感染（lower respiratory tract infections，LRTIs）相关炎症及免疫紊乱的作用机制。方法 基于TCMSP数据库筛选JZ活性成分，运用SysDT算法预测作用靶点。构建药材-成分-靶点网络筛选核心靶标，并通过Metascape平台进行基因本体（gene ontology，GO）及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析。采用AutoDock软件对关键活性成分与枢纽靶点进行分子对接。整合LRTIs患者外周血单核细胞的差异表达基因（differentially expressed genes，DEGs）与JZ预测靶点，筛选共有靶标并进行功能分析。构建脂多糖联合γ干扰素诱导的人单核细胞炎症模型，给予0.67、1.34、2.68 mg/mL JZ干预后进行转录组测序，采用qRT-PCR和流式细胞术验证核心靶点。结果 获得34个活性成分及210个潜在作用靶点，其中83个免疫相关靶标显著富集于肿瘤坏死因子（tumor necrosis factor，TNF）、白细胞介素-17（interleukin-17，IL-17）信号通路（P＜0.05）。294个LRTIs相关DEGs与JZ靶点进行交集，获得11个关键基因，涉及病毒感染、TNF及IL-17信号通路（P＜0.05）。单核细胞炎症模型成功复现66%临床相关DEGs。低、中、高剂量JZ干预分别纠正76.4%、83.1%和88.7%的异常基因表达，并参与调控TNF、核因子-κB（nuclear factor-κB，NF-κB）等通路。qRT-PCR结果显示，JZ呈剂量相关性地下调TNF、IL-6、趋化因子配体2 mRNA表达（P＜0.05、0.01、0.001），显著上调基质金属蛋白酶9和SMAD3 mRNA表达（P＜0.01、0.001）。流式细胞术检测结果证实JZ干预可降低CD86⁺ M1型巨噬细胞比例及TNF-α分泌（P＜0.05、0.01）。结论 JZ通过多靶点协同调控TNF、NF-κB及IL-17信号通路，缓解炎症症状并重塑免疫稳态，从而发挥抗LRTIs作用。

Objective To investigate the mechanism of Jinzhen Oral Liquid (金振口服液, JZ) on ameliorating inflammation and immune dysregulation related to lower respiratory tract infections (LRTIs) using an integrated approach combining systems pharmacology, transcriptomics, molecular docking and experimental validation. Methods Active components of JZ were screened using TCMSP database. Potential targets were predicted using SysDT algorithm. A herb-component-target network was constructed to identify core targets, followed by gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses using Metascape. Molecular docking was performed to key active components and herb targets using AutoDock software. Differentially expressed genes (DEGs) in peripheral blood mononuclear cells from LRTIs patients were integrated with JZ-predicted targets to identify overlapping targets for functional analysis. A model of human monocyte inflammation induced by lipopolysaccharide and γ interferon was constructed, and transcriptome sequencing was performed after the intervention of JZ (0.67, 1.34, 2.68 mg/mL). The expressions of core targets was verified using qRT-PCR and flow cytometry. Results A total of 34 active components and 210 potential targets were identified, of which 83 immune-related targets were significantly enriched in the tumor necrosis factor (TNF) and interleukin-17 (IL-17) signaling pathways (P < 0.05). A total of 294 LRTIs-related DEGs were intersected with JZ targets to obtain 11 key genes, involved in viral infection, TNF and IL-17 signaling pathways (P < 0.05). The monocyte inflammation model successfully recapitulated 66% of clinical DEGs. Low-, medium-, and high-doses of JZ restored 76.4%, 83.1% and 88.7% of dysregulated gene expression, respectively, modulating pathways including TNF and nuclear factor-κB (NF-κB). qRT-PCR demonstrated that JZ dose-dependently down-regulated TNF, IL-6 and chemokine ligand 2 mRNA expressions (P < 0.05, 0.01, 0.001), and significantly up-regulated the mRNA expressions of matrix metalloproteinase 9 and SMAD3 (P < 0.01, 0.001). Flow cytometry confirmed JZ reduced CD86⁺ M1 macrophage proportion and TNF-α secretion (P < 0.05, 0.01). Conclusion JZ exerts anti-LRTIs effects by synergistically regulating TNF, NF-κB and IL-17 signaling pathways through multiple targets to alleviate inflammatory symptoms and remodel immune homeostasis.

R285.5

国家工信部产业基础再造和制造业高质量发展专项（TC2308068）