目的 整合网络药理学、分子对接及体外实验探讨桑枝活性成分桑黄素治疗胃癌的作用机制。方法 通过网络药理学筛选桑枝潜在靶点与胃癌疾病靶点，取交集获得共同靶点；采用蛋白-蛋白相互作用（protein-protein interaction，PPI）网络拓扑分析、基因本体论（gene ontology，GO）与京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析、GEO数据集筛选核心靶点。通过分子对接与分子动力学模拟验证桑枝活性成分桑黄素与磷脂酰肌醇3-激酶调节亚基1（phosphatidylinositol 3-kinase regulatory subunit 1，PIK3R1）的结合能力及复合物稳定性。体外实验以100、200、300、400 μmol/L桑黄素单独或联合磷脂酰肌醇3激酶（phosphatidylinositol 3-kinase，PI3K）激动剂740Y-P干预人胃腺癌AGS细胞，采用CCK-8法、平板克隆形成实验检测细胞增殖能力，流式细胞术检测细胞凋亡率与细胞周期分布，Western blotting检测PI3K/蛋白激酶B（protein kinase B，Akt）通路、凋亡及周期相关蛋白表达。结果 共获得桑枝潜在靶点178个、胃癌相关靶点13 100个，交集靶点159个；富集分析显示关键通路为PI3K/Akt通路。桑黄素与PIK3R1结合亲和力良好，PIK3R1在胃癌组织中显著高表达，复合物结构稳定。体外实验结果显示，桑黄素可剂量相关性抑制AGS细胞增殖，诱导G₀/G₁期阻滞并促进细胞凋亡（P＜0.05、0.01）；同时下调PI3K/Akt通路、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、周期蛋白D1（cyclin D1，CCND1）、细胞周期蛋白依赖性激酶4（cyclin-dependent kinase 4，CDK4）、CDK6蛋白表达（P＜0.05、0.01），上调Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、p21蛋白表达（P＜0.01）；与740Y-P联用后可显著逆转桑黄素的抑增殖、促凋亡及周期阻滞作用（P＜0.05、0.01）。结论 桑黄素可通过靶向并稳定结合PIK3R1，抑制PI3K/Akt通路活化，进而诱导胃癌细胞周期阻滞、促进凋亡并抑制增殖，发挥抗胃癌活性。

Objective To explore the mechanism of morin, an active ingredient of Sangzhi (Mori Ramulus), against gastric cancer based on network pharmacology, molecular docking and in vitro experiments. Methods Potential targets of Mori Ramulus and gastric cancer-related targets were screened by network pharmacology, and the intersection of targets was obtained. Protein-protein interaction (PPI) network topological analysis, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, and GEO dataset validation were used to screen core targets. Molecular docking and molecular dynamics simulation were applied to verify the binding ability of morin (the active ingredient of Mori Ramulus) to phosphatidylinositol 3-kinase regulatory subunit 1 (PIK3R1) and the stability of the formed complex. In in vitro experiments, human gastric adenocarcinoma AGS cells were treated with 100, 200, 300, 400 μmol/L morin alone or combined with phosphatidylinositol 3-kinase (PI3K) agonist 740Y-P. Cell proliferation ability was detected by CCK-8 assay and plate clone formation assay. Cell apoptosis rate and cell cycle distribution were detected by flow cytometry. Expressions of PI3K/protein kinase B (Akt) pathway, apoptosis and cycle related proteins were detected by Western blotting. Results A total of 178 potential targets of Mori Ramulus, 13 100 gastric cancer-related targets, and 159 intersecting targets were identified. Enrichment analysis showed that the key pathway was the PI3K/Akt pathway. Morin had strong binding affinity with PIK3R1, was significantly highly expressed in gastric cancer tissues, and the constructed morin-PIK3R1 complex maintained excellent stability. The in vitro experiment results showed that morin could inhibit AGS cells proliferation in a dose-dependent manner, induce G₀/G₁ phase arrest, and promote cell apoptosis (P < 0.05, 0.01). Meanwhile, morin downregulated the expressions of PI3K/Akt pathway, B-cell lymphoma-2 (Bcl-2), cyclin D1 (CCND1), cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase 6 (CDK6) protein (P < 0.05, 0.01), and upregulated the expressions of Bcl-2 associated X protein (Bax) and p21 (P < 0.01). Combined treatment with 740Y-P significantly reversed the effects of morin on proliferation, apoptosis and cell cycle (P < 0.05, 0.01). Conclusion Morin can exert anti-gastric cancer activity by targeting and stably binding to PIK3R1, inhibiting the activation of PI3K/Akt pathway, inducing cell cycle arrest, promoting apoptosis, and inhibiting proliferation in gastric cancer cells.

R285.5

国家自然科学基金资助项目（81703001）；河北省自然科学基金资助项目（H2026406050）；河北省泛血管疾病重点实验室开放项目（FXGJBKFKT2504）；河北省高等学校科学技术研究项目（QN2026257）；承德医学院大学生创新创业训练计划项目（2025007，2024107）