| 摘要: |
| 目的：采用网络药理学、分子对接技术及细胞实验等方法，系统探究花生衣中金圣草黄素通过磷脂酰肌醇3-激酶-蛋白激酶B-哺乳动物雷帕霉素靶蛋白（phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin，PI3K-AKT-mTOR)通路抑制肝癌的作用机制。方法：利用传统中药系统药理学（ Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform，TCMSP)数据库筛选出花生衣的活性成分，预测肝癌相关靶点，通过基因本体（gene ontology，GO)功能与京都基因和基因组数据库（Kyoto Encyclopedia of Genes and Genomes，KEGG)通路富集分析评估关键靶点涉及的生物过程与信号通路。采用分子对接解析金圣草黄素与PI3K、AKT、mTOR、固醇调节元件结合蛋白1（sterol regulatory element-binding protein 1，SREBP-1)、脂肪酸合酶（ fatty acid synthase，FASN)和硬脂酰辅酶A去饱和酶1（stearoyl-CoA desaturase 1，SCD1)之间的相互作用。通过细胞计数试剂盒-8（Cell Counting Kit-8，CCK-8)进行细胞增殖实验、克隆形成实验及蛋白质印迹（Western blot）分析，验证金圣草黄素对肝细胞癌细胞系G2细胞（hepatocellular carcinoma G2，HepG2）增殖及脂肪酸合成的抑制作用。结果：网络药理学分析筛选出了花生衣的关键靶点，并揭示了PI3K-AKT-mTOR信号通路在金圣草黄素抗肝癌作用中的核心调控地位。GO功能富集分析显示，金圣草黄素主要涉及细胞凋亡、外源性刺激应答及类固醇激素反应等生物过程。KEGG通路富集分析进一步证实PI3K-AKT通路在其抗肝癌中的关键作用。分子对接结果表明，金圣草黄素可与丝氨酸/苏氨酸激酶1、mTOR、SREBP-1、FASN和SCD1形成稳定结合，其结合自由能均＜-5 kcal/mol，揭示其具有较高的结合亲和力。细胞实验结果显示，金圣草黄素能够显著抑制HepG2细胞的增殖，并通过下调磷酸化AKT（p-AKT)、mTOR、SREBP-1、FASN和SCD1的蛋白表达，抑制肝癌细胞的脂肪酸合成，进一步验证了其在PI3K-AKT-mTOR信号通路中的调控作用。结论：金圣草黄素通过调控PI3K-AKT-mTOR信号通路，抑制肝癌细胞的增殖与脂质代谢，揭示了其作为花生衣中潜在抗肝癌成分的药理机制。 |
| 关键词: 肝癌 金圣草黄素 网络药理学 分子对接 PI3K-AKT-mTOR通路 脂肪酸合成 |
| DOI： |
|
|
| Nvestigating the Mechanism of Calycosin in Inhibiting Hepatocellular Carcinoma Cell Proliferation and Lipid Metabolism Based on Network Pharmacology and Experimental Studies |
| ZHANG Chenbin,YIN Xin,SU Bowu |
| (Hunan Normal University，Changsha 410018，Hunan，China) |
| Abstract: |
| Objective：To investigate the mechanism of action of chrysoeriol，an active component from peanut skin，in inhibiting liver cancer through the phosphatidylinositol 3-kinase （PI3K)/protein kinase B （AKT)/mechanistic target of rapamycin （mTOR) pathway based on network pharmacology，molecular docking，and cell experiments.Methods：Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database was used to obtain the active components of peanut skin，and the targets associated with liver cancer were predicted.The gene ontology （GO) functional enrichment analysis and the Kyoto Encyclopedia of Genes and Genomes （KEGG) pathway enrichment analysis were performed to identify the biological processes and signaling pathways involving the key targets.Molecular docking was used to analyze the interaction between chrysoeriol and PI3K/AKT/mTOR/sterol regulatory element-binding protein 1 （SREBP-1)/fatty acid synthase （FASN)/stearoyl-CoA desaturase 1 （SCD1).CCK-8 assay，colony formation assay，and Western blot were used to validate the inhibitory effect of chrysoeriol on HepG2 cell proliferation and fatty acid synthesis.Results：The key targets of peanut skin were obtained by network pharmacology，which also revealed the core regulatory role of the PI3K-AKT-mTOR signaling pathway in the effect of chrysoeriol against liver cancer.The GO functional enrichment analysis showed that chrysoeriol was mainly involved in the biological processes such as cell apoptosis，response to external stimuli，and steroid hormone responses，and the KEGG pathway enrichment analysis further confirmed the key role of the PI3K-AKT pathway in the treatment of liver cancer.Molecular docking showed stable binding of chrysoeriol to serine/threonine kinase 1，mTOR，SREBP-1，FASN，and SCD1，with a binding free energy of <-5 kcal/mol，suggesting that chrysoeriol had high affinity.Cell experiments showed that chrysoeriol significantly inhibited HepG2 cell proliferation and played a regulatory role in the PI3K-AKT-mTOR signaling pathway by downregulating the protein expression of phosphorylated AKT，mTOR，SREBP-1，FASN，and SCD1 and inhibiting fatty acid synthesis in hepatoma cells.Conclusion：Chrysoeriol inhibits hepatoma cell proliferation and lipid metabolism by regulating the PI3K-AKT-mTOR signaling pathway，which reveals the pharmacological mechanism of chrysoeriol as a potential therapeutic agent against liver cancer derived from peanut skin. |
| Key words: liver cancer chrysoeriol network pharmacology molecular docking phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin pathway fatty acid synthesis |
