尽管近年来小檗碱的治疗作用受到了一些关注，但其抗胃癌（SC）的潜在机制仍不清楚。在本研究中，我们旨在利用网络药理学和实验验证方法阐明小檗碱对抗SC作用的机制。使用几个公开数据库来收集小檗碱和SC的靶点。基于小檗碱对抗 SC 的潜在靶点进行蛋白质-蛋白质相互作用 (PPI) 网络、富集分析和分子对接。通过使用列线图和受试者工作特征（ROC）分析来预测目标的潜在临床意义和预后价值。然后测定小檗碱处理的SC细胞的活力和凋亡。此外，还进行了活性氧（ROS）、线粒体膜电位（MMP）和三磷酸腺苷（ATP）测量以及蛋白质印迹分析来验证预测的机制。确定了小檗碱对抗SC的76个潜在靶点。 PPI网络的构建使得能够识别AKT1、TP53、IL6、JUN和MAPK1等枢纽目标。富集分析表明小檗碱参与细胞凋亡、线粒体自噬、ROS代谢过程、AMPK和MAPK信号通路。 hub靶标的表达水平也有助于SC患者的临床预后。分子对接揭示了小檗碱与靶蛋白（包括 AMPK、TP53 和 MAPK1）之间直接相互作用的可能模式。实验结果表明，小檗碱降低 SC 细胞活力，促进细胞凋亡和 ROS 生成，并有助于降低 MMP 和 ATP 水平。 Western blot检测表明小檗碱增加AMPK和TP53的表达，同时降低磷酸化-MAPK3/1的表达。我们利用网络药理学方法阐明了小檗碱对抗SC的潜在作用机制。基于实验方法验证了一些预测的抗 SC 作用机制。我们的研究结果为小檗碱作为细胞凋亡诱导剂和能量代谢调节剂对抗 SC 提供了有意义的基础。但体内实验和临床研究还需进一步开展。此外，有必要深入研究小檗碱的潜在负面影响。2024 AME Publishing Company。版权所有。

Although the therapeutic effects of berberine have received some attention in recent years, its potential mechanisms underlying its action against stomach carcinoma (SC) remain unclear. In this study, we aimed to elucidate the mechanisms underlying the effects of berberine against SC using a network pharmacology and experimental verification approach.Several publicly available databases were used to collect the targets of berberine and SC. Protein-protein interaction (PPI) network, enrichment analyses and molecular docking were performed based on the potential targets of berberine against SC. The potential clinical significance and prognostic value of the targets were predicted by using nomogram and receiver operating characteristic (ROC) analyses. Then the viability and apoptosis of SC cells treated with berberine were determined. Moreover, reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) measurements and western blot assay were carried out to validate the predicted mechanisms.Seventy-six potential targets of berberine against SC were identified. The construction of PPI network enabled the identification of hub targets, such as AKT1, TP53, IL6, JUN and MAPK1. Enrichment analyses showed that berberine was involved in apoptosis, mitophagy, ROS metabolic process, AMPK and MAPK signaling pathway. The expression levels of hub targets also contributed to the clinical prognosis of patients with SC. Molecular docking revealed the possible patterns of direct interaction between berberine and target proteins, including AMPK, TP53 and MAPK1. Experimental results showed that berberine reduced SC cell viability, promoted apoptosis and ROS generation, and contributed to reductions in MMP and ATP levels. Western blot assay demonstrated that berberine increased AMPK and TP53 expression, while decreased phosphorylated-MAPK3/1 expression.We elucidated the potential action mechanisms of berberine against SC using a network pharmacology approach. Some predicted mechanisms underlying the anti-SC effects were verified based on experimental approaches. Our findings provide a meaningful foundation for berberine as a cellular apoptosis-inducing and energy metabolism-regulating agent against SC. However, in vivo experiments and clinical studies need to be further carried out. Moreover, it is necessary to study the potential negative effects of berberine thoroughly.2024 AME Publishing Company. All rights reserved.