目的 采用网络药理学结合分子对接及体内验证，探究鸦胆子Brucea javanica治疗结直肠癌的活性成分、关键作用靶点及潜在的分子机制。方法 从公共数据库中检索并收集鸦胆子的活性成分和对应的靶点以及结直肠癌相关的靶点，通过网络药理学分析出鸦胆子治疗结直肠癌的活性成分、成分-疾病的交集靶点以及可能的信号通路。通过分子对接预测活性成分与靶点蛋白的结合能力。通过体内实验进一步验证鸦胆子活性成分治疗结直肠癌的作用和作用机制。结果 在满足筛选条件的15个成分中共筛出鸦胆子苦醇、木犀草素、鸦胆子苷B、β-谷甾醇4个鸦胆子生物活性成分及32个鸦胆子治疗结直肠癌的作用靶点，表皮因子生长受体（epidermal growth factor receptor，EGFR）、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）及细胞周期蛋白D1（cyclin D1）是鸦胆子治疗结直肠癌的关键靶点，EGFR/磷脂酰肌醇3-激酶（phosphatidylinositol-3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）信号通路可能在鸦胆子治疗结直肠癌中发挥重要作用。分子对接结果表示4个生物活性成分均可较好结合，其中鸦胆子苦醇与3个关键靶点的平均结合能负值最大，结合最稳定，可能是鸦胆子抗结直肠癌最有效的活性成分。动物实验结果表明，与对照组比较，鸦胆子苦醇明显抑制裸鼠体内移植瘤的体积和质量（P＜0.01），下调肿瘤组织中EGFR、PI3K和Akt的蛋白表达水平（P＜0.01），下调与G₁/G₀期细胞阻滞相关的cyclin D1和细胞周期蛋白依赖性激酶4（cyclin-dependent kinase 4，CDK4）的表达（P＜0.01），上调与肿瘤细胞凋亡相关的cleaved Caspase-3表达及B淋巴细胞瘤-2相关X蛋白（B-cell lymphoma-2 associated X protein，Bax）/B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）值（P＜0.01），下调与迁移和侵袭相关的蛋白基质金属蛋白酶9（matrix metalloproteinase 9，MMP9）、MMP7表达（P＜0.01）；鸦胆子苦醇组裸鼠肝功能、肾功能及血液指标均未见显著性差异。结论 鸦胆子能够通过多靶点、多通路治疗结直肠癌，其主要活性成分为鸦胆子苦醇，其机制可能与抑制EGFR/PI3K/Akt信号通路从而引起G₁/G₀期阻滞，抑制增殖、侵袭和迁移，促进细胞凋亡有关。

Objective To explore the active components, key targets and potential mechanisms of Brucea javanica in treatment of colorectal cancer by network pharmacology combined with molecular docking and in vivo verification. Methods The active components, corresponding targets and colorectal cancer-related targets of B. javanica were retrieved and collected from the public database. The active components, component-disease intersection targets and possible signal pathways of B. javanica in treatment of colorectal cancer were analyzed through network pharmacology. Binding ability of active components to target protein was predicted by molecular docking. The effect and mechanism of B. javanica active components in treatment of colorectal cancer were further verified by in vivo experiments. Results Among the 15 components that met the screening conditions, four bioactive components (brusatol, luteolin, bruceoside B, β-sitosterol) and 32 therapeutic targets of B. javanica were screened. The epidermal growth factor receptor (EGFR), cystein-asparate protease-3 (Caspase-3) and cyclin D1 were the key targets of B. javanica in the treatment of colorectal cancer. EGFR/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway may played an important role of B. javanica in treatment of colorectal cancer. The results of molecular docking showed that all four bioactive components could bind well, and average binding energy of brusatol with three key targets was the most negative and the binding was the most stable, which may be the most effective active component of B. javanica against colorectal cancer. The results of animal experiments showed that compared with control group, brusatol significantly inhibited the volume and weight of transplanted tumor in nude mice (P < 0.01), down-regulated the protein expression levels of EGFR, PI3K and Akt in tumor tissues (P < 0.01), down-regulated cyclin D1 and cyclin-dependent kinase 4 (CDK4) expressions (P < 0.01), up-regulated the expressions of tumor cell apoptosis related protein such as cleaved Caspase-3 and B-cell lymphoma-2 associated X protein (Bax)/B-cell lymphoma-2 (Bcl-2) (P < 0.01), down-regulated the expressions of migration and invasion related proteins such as matrix metalloproteinase 9 (MMP9) and MMP7 (P < 0.01); There was no significant difference in liver function, renal function and blood indexes of nude mice in brusatol group. Conclusion B. javanica can treat colorectal cancer through multiple targets and channels, and its main active component is bruceol. Its mechanism may be related to inhibiting EGFR/PI3K/Akt signaling pathway, thus causing G₁/G₀ phase block, inhibiting proliferation, invasion and migration, and promoting cell apoptosis.

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吉林省发展和改革委员会项目（2014G073）；吉林省直厅局项目（2019SCZT040）；吉林省科技厅基础处项目（20200201589JC）