目的 基于网络药理学和体外实验探讨白藜芦醇抗横纹肌肉瘤的分子作用机制。方法 综合运用网络药理学、分子对接及分子动力学模拟筛选并验证白藜芦醇抗横纹肌肉瘤的核心靶点；使用基因表达综合数据库（GEO）数据集对核心靶点进行差异表达、生存分析和基因集富集分析（GSEA）通路富集分析。体外培养人横纹肌肉瘤A-204和RD细胞株，采用MTT法、流式细胞术检测不同浓度白藜芦醇对细胞活力和凋亡的影响，Western blotting检测肿瘤蛋白53（p53）及其磷酸化形式（p-p53）的表达水平。结果 网络药理学分析显示，肿瘤抑制因子p53（TP53）、蛋白激酶B1（Akt1）、信号转导及转录激活因子3（STAT3）、连环蛋白β-1（CTNNB1）、转录因子Jun（JUN）为核心靶点，分子对接及动力学模拟表明白藜芦醇与TP53结合稳定；转录组分析显示，TP53在横纹肌肉瘤组织中异常表达且与患者生存预后相关，同时p53信号通路显著富集。体外实验表明，白藜芦醇呈剂量相关性抑制横纹肌肉瘤细胞增殖并诱导凋亡；Western blotting结果显示白藜芦醇未改变总p53蛋白水平，但显著上调p-p53表达（P＜0.01、0.001）。结论 白藜芦醇主要通过激活p53磷酸化发挥抗横纹肌肉瘤作用，为横纹肌肉瘤的新型治疗策略提供实验依据。

Objective To explore the molecular mechanism of resveratrol in resisting rhabdomyosarcoma based on network pharmacology and in vitro experiments. Methods Network pharmacology, molecular docking, and molecular dynamics simulation were comprehensively employed to screen and validate the core targets of resveratrol against rhabdomyosarcoma. Based on GEO datasets, differential expression, survival analysis, and Gene Set Enrichment Analysis (GSEA) pathway enrichment were performed on the core targets. Human RMS A-204 and RD cell lines were cultured in vitro. MTT assay and flow cytometry were used to detect the effects of different concentrations of resveratrol on cell viability and apoptosis. Western blotting was used to detect the expression levels of Tumor protein 53 (p53) and its phosphorylated form (p-p53). Results Network pharmacology analysis revealed that tumor suppressor factor p53 (TP53), protein kinase B1 (Akt1), signal transducer and activator of transcription 3 (STAT3), catenin β-1 (CTNNB1), and transcription factor Jun (JUN) were the core targets. Molecular docking and kinetic simulation indicated that resveratrol bound stably to TP53. Transcriptome analysis showed that TP53 was abnormally expressed in rhabdomyosarcoma tissues and was related to the patient's survival prognosis. In vitro experiments demonstrated that resveratrol inhibited the proliferation of rhabdomyosarcoma cells in a dose-dependent manner and induced apoptosis. Western blotting results showed that resveratrol did not change the total p53 protein level, but significantly upregulated p-p53 expression (P < 0.01, 0.001). Conclusion Resveratrol exerts its anti-rhabdomyosarcoma effect mainly by activating p53 phosphorylation, providing an experimental basis for novel therapeutic strategies for rhabdomyosarcoma.

R285.5

孝感市自然科学计划项目（XGKJ2025010021）