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目的：明确笔管草醇提物（Ethanol extract of Equisetum debile Roxb.,EED）防治高脂血症的核心药效成分，揭示其“成分-靶点-通路”的作用机制，为笔管草防治高脂血症的临床应用与新药研发提供科学依据。方法：采用油酸诱导HepG2细胞建立脂质蓄积模型，通过油红O染色验证EED的降脂活性；运用超高效液相色谱-串联四级杆飞行时间质谱结合PubChem、TCMSP等在线数据库，系统鉴定EED灌胃给药后的大鼠入血成分及代谢产物；借助Swiss Target Prediction数据库筛选入血成分与高脂血症的交集靶点，利用STRING平台构建靶点蛋白互作网络，通过DAVID数据库进行KEGG信号通路富集分析；采用AutoDock Vina软件进行分子对接，结合PyMOL可视化分析潜在活性成分与关键靶点的结合模式及亲和力；为了进一步验证EED对关键基因表达的调控作用，采用RT-qPCR技术检测HepG2细胞在给药前后STAT3、HSP90AA1、EGFR、PTGS2及ESR1等基因的mRNA水平变化。结果：EED可浓度依赖性降低油酸诱导的HepG2细胞内TG含量及脂质堆积（P<0.05）；从大鼠血清中初步鉴定出EED的17个主要入血成分，包括柚皮素、山奈酚等黄酮类成分；网络药理学分析显示，EED核心入血成分柚皮素、山奈酚、金合欢素、槲皮素等可共同作用于STAT3、AKT1、EGFR、HSP90AA1等31个高脂血症相关核心靶点；KEGG富集分析提示上述靶点主要富集于脂质与动脉粥样硬化通路等15条核心通路；分子对接结果证实，PTGS2与棉花素、PTGS2与山奈酚、PTGS2与甘草素，以及ESR1与甘草素的结合自由能均低于-9.0 kcal/mol，结合模式稳定；RT-qPCR检测结果显示，模型组中STAT3、HSP90AA1、EGFR、PTGS2基因的mRNA表达水平上调，而ESR1基因的mRNA表达下调，EED干预后可逆转上述基因的表达异常。结论：EED通过“黄酮类成分集群-核心靶点-脂代谢相关通路”发挥防治高脂血症功效，其核心药效物质为甘草素、山奈酚、棉花素等黄酮类成分，脂质与动脉粥样硬化信号通路可能是其关键作用通路。
Abstract：Objective : To identify the core active constituents of the ethanol extract of(Ethanol extract of Equisetum debile Roxb.,EED) responsible for its therapeutic effects against hyperlipidemia, and to elucidate its mechanism of action via the "constituents-targets-pathways" axis, thereby providing a scientific basis for its clinical application and new drug development. Methods : An in vitro model of lipid accumulation was established using oleic acid-induced HepG2 cells. The lipid-lowering efficacy of EED was verified via Oil Red O staining. Systemic identification of the absorbed constituents and metabolites of EED in rat serum following oral administration was performed using UPLC-Q-TOF-MS/MS, assisted by online databases including PubChem and TCMSP. Potential targets of these serum constituents related to hyperlipidemia were screened using the Swiss Target Prediction database. A protein-protein interaction(PPI) network was constructed using the STRING platform, and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis was conducted via the DAVID database. Molecular docking was performed using AutoDock Vina, and the binding modes and affinities between potential active constituents and key targets were visualized and analyzed with PyMOL. To further verify the regulatory effect of EED on the expression of key genes, the RT-qPCR technique was used to detect the changes in the mRNA levels of genes such as STAT3, HSP90 AA1, EGFR, PTGS2, and ESR1 in HepG2 cells before and after drug administration. Results : EED concentration-dependently reduced both intracellular TG content and lipid droplet accumulation in oleic acid-induced HepG2 cells(P<0. 05). Seventeen major absorbed constituents, primarily flavonoids such as naringenin and kaempferol, were identified in rat serum. Network pharmacology analysis revealed that 31 core hyperlipidemia-related targets, including STAT3, AKT1, EGFR, and HSP90 AA1, were commonly modulated by key serum constituents like naringenin, kaempferol, acacetin, and quercetin. KEGG enrichment analysis indicated that these targets were significantly enriched in 15 core pathways, predominantly the Lipid and Atherosclerosis signaling pathway. Molecular docking confirmed strong binding affinities, with binding free energies below-9. 0 kcal/mol for the complexes of PTGS2 with acacetin, PTGS2 with kaempferol, PTGS2 with liquiritigenin, and ESR1 with liquiritigenin, indicating stable binding modes. RT-qPCR detection results showed that the mRNA expression levels of STAT3, HSP90 AA1, EGFR, and PTGS2 genes were significantly increased in the model group, while the expression of the ESR1 gene was significantly decreased. After EED intervention, the expression changes of these genes were all reversed. Conclusion : EED exerts its anti-hyperlipidemic effect through the synergistic interactions of a "flavonoid cluster-core targets-lipid metabolism-related pathways" network. The core pharmacodynamic substances are flavonoids, including liquiritigenin, kaempferol, and acacetin, with the PTGS2 and ESR1 signaling pathways potentially serving as critical mechanistic routes.
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基本信息:
中图分类号:R285.5
引用信息:
[1]石雪颖,申福葵,魏志豪,等.笔管草醇提物防治高脂血症的潜在药效物质及机制研究[J].赣南医科大学学报,2026,46(05):410-420.
基金信息:
江西省自然科学基金项目（20252BAC200578,20253BAC280096）; 江西省职业早期青年科技人才项目（20252BEJ730276）; 赣州市科技计划校地合作项目（2025XDCE0014,2025XDCE0032）
2025-11-21
2025
2026-05-21
2026-01-31
2026
1
2026-05-22
2026-05-22
2026-05-22