Lonicerae japonicae flos (LJF) is widely used for the treatment of inflammation-related diseases in traditional Chinese medicine (TCM). To clarify the anti-inflammatory mechanism of LJF, 29 compounds with high content in LJF were selected for network pharmacology. Then, a comprehensive network pharmacology strategy was implemented, which involved compound-inflammation-target construction, protein-protein interaction (PPI) network analysis, and enrichment analysis. Finally, molecular docking and in vitro experiments were performed to verify the anti-inflammatory activity and targets of the key compound. As a result, 279 inflammation-associated proteins were identified, which are mainly involved in the AGE/RAGE signaling pathway in diabetic complications, the HIF-1 signaling pathway, the PI3K-AKT signaling pathway, and EGFR tyrosine kinase inhibitor resistance. A total of 12 compounds were linked to more than 35 targets, including apigenin, kaempferol, quercetin, luteolin, and ferulic acid. The results of molecular docking showed that AKT has the most binding activity, exhibiting certain binding activity with 10 compounds, including vanillic acid, protocatechuic acid, secologanic acid, quercetin, and luteolin; the results of qRT-PCR and WB confirmed that two key compounds, secologanic acid and luteolin, could significantly decrease the secretion of TNF-α and the AKT expression of RAW264.7 murine macrophages stimulated by LPS (lipopolysaccharide). These results demonstrate that the comprehensive strategy can serve as a universal method to illustrate the anti-inflammatory mechanisms of traditional Chinese medicine by identifying the pathways or targets.

中文翻译：

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基于网络药理学的金银花抗炎作用潜在靶点鉴定

金银花(LJF) 在中医 (TCM) 中广泛用于治疗炎症相关疾病。为阐明LJF的抗炎机制，选取29个LJF中含量较高的化合物进行网络药理学。然后，实施了综合网络药理学策略，包括化合物-炎症-靶点构建、蛋白质-蛋白质相互作用（PPI）网络分析和富集分析。最后，分子对接和体外进行了实验以验证关键化合物的抗炎活性和靶点。结果鉴定出279个炎症相关蛋白，主要参与糖尿病并发症的AGE/RAGE信号通路、HIF-1信号通路、PI3K-AKT信号通路和EGFR酪氨酸激酶抑制剂耐药。共有 12 种化合物与超过 35 个靶标相关联，包括芹菜素、山柰酚、槲皮素、木犀草素和阿魏酸。分子对接结果表明AKT的结合活性最强，与香草酸、原儿茶酸、西洛木酸、槲皮素、木犀草素等10种化合物具有一定的结合活性；qRT-PCR 和 WB 的结果证实了两种关键化合物，西洛木酸和木犀草素，α和 LPS（脂多糖）刺激的 RAW264.7 鼠巨噬细胞的 AKT 表达。这些结果表明，综合策略可以作为一种通用方法，通过识别途径或靶点来说明中药的抗炎机制。