目的 基于网络药理学、转录组学与动物实验探究小儿风热清合剂（口服液）治疗急性肺炎的作用机制。方法 通过网络药理学筛选小儿风热清合剂（口服液）的活性成分及作用靶点，与急性肺炎相关靶点取交集，构建蛋白质相互作用（PPI）网络，并进行基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路富集分析，通过分子对接验证活性成分与核心靶点的结合能力。采用脂多糖（LPS）气管滴注法建立小鼠急性肺损伤模型，通过苏木精–伊红（HE）染色观察肺组织病理学改变，利用实时荧光定量PCR（qRT-PCR）检测肺组织中白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）mRNA的表达水平。通过转录组测序（RNA-seq）分析小儿风热清合剂（口服液）干预后的差异表达基因（DEGs）与核心信号通路，与网络药理学预测靶点交叉验证。结果 网络药理学筛选获得148个小儿风热清合剂（口服液）治疗急性肺炎的交集靶点，PPI网络分析显示信号转导及转录激活蛋白3（STAT3）、p53肿瘤抑制蛋白（TP53）、热休克蛋白90α家族A类成员1（HSP90AA1）等为核心靶点，富集于晚期糖基化终末产物–受体（AGE-RAGE）信号通路、白细胞介素-17（IL-17）通路和丝裂原活化蛋白激酶（MAPK）等信号通路。分子对接结果显示，小儿风热清合剂（口服液）中的青黛酮、乔松素、黄芩素、β-胡萝卜素、菜豆素及圣草酚等6种活性成分与丝裂原活化蛋白激酶10（MAPK10）有良好的结合能力。动物实验显示，小儿风热清合剂（口服液）可明显减轻LPS诱导的急性肺损伤小鼠肺泡壁增厚、炎症细胞浸润及肺泡出血等病理损伤，并显著下调肺组织中IL-6、IL-1β、TNF-α mRNA的表达水平（P＜0.01、0.001）。转录组学联合网络药理学筛选出MAPK10/c-Jun氨基末端激酶3（JNK3）为潜在关键靶点。Western blotting结果显示小儿风热清合剂（口服液）可以降低肺组织JNK、p-JNK蛋白表达水平（P＜0.05、0.01）。结论 小儿风热清合剂（口服液）可能通过上述活性成分作用于MAPK10/JNK3关键靶点，抑制JNK信号通路活化，减少促炎因子表达，从而改善LPS诱导的小鼠急性肺炎肺组织损伤。

Objective To investigate potential mechanism of Xiao’er Fengre Qing Mixture (Oral Liquid) on acute pneumonia by integrating network pharmacology, transcriptomics, and animal experiments. Methods Network pharmacology was used to screen the active components and corresponding targets of Xiao’er Fengre Qing Mixture (Oral Liquid), and the overlapping targets with acute pneumonia-related genes were obtained. A PPI network was constructed, followed by GO and KEGG enrichment analysis. Molecular docking was employed to verify the binding affinity between active components and the core target. An LPS-induced acute lung injury mice model was established by intratracheal instillation. Lung pathological changes were observed by HE staining. The mRNA expression levels of IL-6, IL-1β, and TNF-α in lung tissues were detected using qRT-PCR. RNA-seq was performed to analyze the DEGs and core signaling pathways following Xiao’er Fengre Qing Mixture (Oral Liquid) intervention. The DEGs were cross-validated with network pharmacology-predicted targets to screen for core targets. Results Network pharmacology identified 148 overlapping targets of Xiao’er Fengre Qing Mixture (Oral Liquid) for pneumonia treatment. PPI network analysis revealed that STAT3, TP53, and HSP90AA1 were core targets. These targets were mainly enriched in the AGE-RAGE, IL-17, and MAPK signaling pathways. Molecular docking confirmed that flavonoid components of Xiao’er Fengre Qing Mixture (Oral Liquid), including qingdainone, pinocembrin, baicalein, β-carotene, phaseol and eriodictyol, exhibited favorable predicted binding to MAPK10. Xiao’er Fengre Qing Mixture (Oral Liquid) significantly alleviated LPS-induced lung pathological damage in mice, including alveolar wall thickening, inflammatory cell infiltration, and alveolar hemorrhage, and significantly reduce expression of IL-6, IL-1β, and TNF-α mRNA (P < 0.01, 0.001). Integration of transcriptomic data with network pharmacology identified MAPK10/JNK3 as a potential key target. Western blotting results showed that Xiao’er Fengre Qing Mixture (Oral Liquid) reduced the protein expression levels of JNK and p-JNK in lung tissue (P < 0.05, 0.01). Conclusion Xiao’er Fengre Qing Mixture (Oral Liquid) may act through the above-mentioned active components on the key target MAPK10/JNK3 to inhibit JNK signaling pathway activation, reduce pro-inflammatory cytokine expression, and thereby ameliorate LPS-induced lung tissue injury in mice with acute pneumonia.

R286.4

广西大学高层次人才科研启动基金项目（ZX01080033424004）；中央引导地方科技发展资金项目（264Z2501G）