目的 探讨仙鹤草抑制肝癌细胞（HCC）的作用及相关机制。方法 TCMSP、UniProt、GeneCards、OMIM、TTD及STRING数据库筛选仙鹤草作用于肝癌的潜在核心靶点；利用UALCAN数据库分析筛选在肝癌组织与正常肝组织中mRNA及蛋白水平高表达的核心靶点；通过Kaplan-Meier Plotter数据库评估其与肝癌患者总体生存期（OS）的相关性，确定预后不良相关的高表达核心靶点。HPA数据库验证所筛选靶点蛋白在正常和肝癌组织中的差异蛋白质表达水平；同时利用UALCAN和Kaplan-Meier Plotter数据库对所筛选靶点在肝癌不同临床分期中的表达水平及其与无进展生存期（PFS）、无复发生存期（RFS）、疾病特异性生存期（DSS）的关系进行验证；进一步通过AutoDock等软件将仙鹤草活性成分与靶点蛋白进行分子对接，利用Pymol软件对结果进行可视化分析。通过体外细胞实验检测仙鹤草对HepG2细胞的增殖、凋亡和周期分布的影响，Westem blotting和Real-time PCR检测MAPK3、CDK2、CDK4、分泌型磷蛋白1（SPP1）在mRNA和蛋白水平的表达变化。结果 生物信息学分析筛选出MAPK3、CDK2、CDK4、SPP1为核心靶点。这些靶点在肝癌组织中mRNA及蛋白表达均显著高于正常组织（P＜0.05），且与患者不良预后显著相关（HR>1，P＜0.05）。分期及生存分析进一步证实，上述靶点在肝癌临床分期中呈持续高表达。CDK2、CDK4、SPP1高表达PFS、RFS及DSS均显著负相关（P＜0.05），MAPK3高表达则与疾病特异性生存期缩短显著相关（P＜0.05）。分子对接分析表明，仙鹤草活性成分与MAPK3、CDK2、CDK4、SPP1靶点均有较强的结合潜力。细胞实验显示，仙鹤草干预后HepG2细胞发生G0/G1和G2/M 期阻滞，集落形成率下降，凋亡率增加（P＜0.05）；MAPK3、CDK2、CDK4、SPP1的mRNA表达均降低；细胞外信号调节激酶1（ERK1）、磷酸化ERK1（p-ERK1）、CDK2、SPP1及CDK4（低剂量组）蛋白水平表达下降，高剂量组磷酸化CDK2（p-CDK2）蛋白亦下降（均P＜0.05）。结论 仙鹤草通过下调MAPK3、CDK2、CDK4、SPP1表达，抑制肝癌细胞增殖并诱导凋亡。

Objective To investigate the effect and mechanism by which agrimonia pilosa inhibits hepatocellular carcinoma (HCC). Methods Potential core targets of agrimonia pilosa active components against HCC were screened using TCMSP, UniProt, GeneCards, OMIM, TTD, and STRING databases. Core targets with elevated mRNA and protein expression levels in HCC tissue versus normal liver tissue were identified via the UALCAN database. Their correlation with overall survival (OS) in HCC patients was assessed using the Kaplan-Meier Plotter database to determine high-expression targets associated with poor prognosis. Differential protein expression of the selected targets between normal and HCC tissue was validated using the HPA database. Meanwhile, their expression levels across different clinical stages of HCC and their associations with progression-free survival (PFS), recurrence-free survival (RFS), and disease-specific survival (DSS) were further verified with UALCAN and Kaplan-Meier Plotter. Molecular docking between agrimonia pilosa active components and target proteins was performed using software such as AutoDock, and the result  were visualized by PyMOL. Cell experiments in vitro were conducted to evaluate the effects of agrimonia pilosa treatment on proliferation, apoptosis, cell cycle distribution, and the mRNA and protein expression levels of MAPK3, CDK2, CDK4, and secreted phosphoprotein 1（SPP1）in HepG2 cells were detecting by Western blotting and Real-time PCR.Results Bioinformatics analysis identified MAPK3, CDK2, CDK4, and SPP1 as core targets. The mRNA and protein expression of these targets was significantly higher in HCC tissue than those in normal tissue (P<0.05), and their high expression was significantly associated with poor prognosis (HR>1, P<0.05). Stage and survival analyses further confirmed that these targets were persistently overexpressed across the clinical stages of HCC. High expression of CDK2, CDK4, and SPP1 was significantly negatively correlated with PFS, RFS, and DSS (P<0.05), while high MAPK3 expression was significantly correlated with shortened DSS (P<0.05). Molecular docking analysis indicated that the active components of agrimonia pilosa possessed strong binding potential to MAPK3, CDK2, CDK4, and SPP1. Cell experiments showed that agrimonia pilosa treatment induced G0/G1 and G2/M phase arrest, decreased the colony formation rate, and increased the apoptosis rate in HepG2 cells (P<0.05). The mRNA expression of MAPK3, CDK2, CDK4, and SPP1 decreased. The protein expression level of extracellular signal-regulated kinase 1（ERK1）, phosphorylated ERK1（p-ERK1）, CDK2, SPP1, and CDK4 (in the low-dose group) was downregulated, and phosphorylated CDK2（p-CDK2）protein in the high-dose group also decreased (P<0.05). Conclusion Agrimonia pilosa inhibits HCC cell proliferation and induces apoptosis by downregulating the expression of MAPK3, CDK2, CDK4, and SPP1.