目的::探究牛耳枫‑辣蓼（Daphniphyllum calycinum‑Polygonum hydropiper, DCPH）对胃黏膜损伤的保护作用及机制。 方法::网络药理学获取DCPH干预胃黏膜损伤的活性成分及主要靶点，构建PPI网络，进行GO和KEGG富集分析筛选可能涉及的生物过程和信号通路。利用乙醇刺激GES‑1细胞构建胃黏膜损伤的体外模型，通过检测细胞活力、活性氧（ROS）含量、TNF‑α和IL‑1β表达、线粒体膜电位水平以及TLR4和NF‑κB蛋白表达，验证DCPH对胃黏膜损伤的保护作用及机制。 结果::通过网络药理学筛选出DCPH干预胃黏膜损伤的潜在活性成分44种，核心靶点TNF、BCL2、EGFR、TLR4等29个，涉及PI3K‑Akt、HIF‑1、NF‑κB和ROS等多个生物过程和信号通路。体外实验结果显示，与正常组相比，乙醇能明显抑制GES‑1细胞活力（P<0.001）、提高ROS含量（P<0.001）、诱导炎性因子TNF‑α和IL‑1β mRNA表达（P<0.001），明显降低细胞内线粒体膜电位，并明显上调TLR4和NF‑κB蛋白表达（P<0.01）。与模型组比较，DCPH明显提高乙醇诱导的GES‑1细胞活力（P<0.05），改善细胞形态，同时降低细胞内ROS水平（P<0.05）、抑制TNF‑α和IL‑1β表达（P<0.05）、恢复线粒体膜电位水平，明显抑制TLR4和NF‑κB蛋白表达（P<0.05）。 结论::DCPH通过下调TLR4/NF‑κB信号通路表达，降低细胞ROS含量和抑制炎症因子表达，恢复细胞线粒体膜电位水平，减轻GES‑1细胞损伤，从而发挥抗胃黏膜损伤作用。

Objective::To explore the protective effects and mechanisms of Daphniphyllum calycinum‑Polygonum hydropiper （DCPH） on gastric mucosal injury. Methods::The active components and main targets of DCPH in the intervention of gastric mucosal injury were obtained from network pharmacology， PPI network was constructed， and GO and KEGG enrichment analysis were carried out to screen possible biological processes and signaling pathways. The model of gastric mucosal injury in vitro was established by stimulating GES‑1 cells with ethanol， and the protective effects and mechanism of DCPH on GES‑1 cells were verified by detecting cell viability， reactive oxygen species （ROS） content， TNF‑α and IL‑1β expression， mitochondrial membrane potential level， and TLR4 and NF‑κB protein expressions. Results::Through network pharmacology， 44 potential active components of DCPH in the intervention of gastric ulcer/gastritis were screened out， including 29 core targets such as TNF， BCL2， EGFR and TLR4， which involved many biological processes and signaling pathways， such as PI3K‑Akt， HIF‑1， NF‑κB and ROS. The in vitro experimental results showed that compared to the normal group， ethanol could significantly inhibit the activity of GES‑1 cells （P<0.001）， increase the ROS content （P<0.001）， induce the expression of inflammatory factors TNF‑α and IL‑1β mRNA （P<0.001）， significantly reduce the intracellular mitochondrial membrane potential， and significantly up‑regulate TLR4 and NF‑κB protein expressions. Compared to the model group， DCPH significantly increased the vitality of GES‑1 cells induced by ethanol （P<0.05）， improved the cell morphology， decreased the intracellular ROS level （P<0.05）， inhibited the expressions of TNF‑α and IL‑1β （P<0.05）， restored the mitochondrial membrane potential level， and significantly inhibited the expressions of TLR4 and NF‑κB proteins （P<0.05）. Conclusion::DCPH alleviates the damage of GES‑1 cells and plays an anti‑gastric mucosal injury role by down‑regulating the expression of TLR4/NF‑κB signaling pathway， reducing the ROS content， inhibiting the expression of inflammatory factors， and restoring the level of mitochondrial membrane potential.

国家自然科学基金（82260920，82474456）；大学生创新创业训练计划项目（S202411810030）