目的 基于血清药物化学、网络药理学及分子动力学模拟探究萆薢分清丸潜在活性成分、治疗慢性肾衰的关键靶点和作用机制。方法 采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱（UHPLC-Q-Orbitrap HRMS）进行萆薢分清丸体内外成分分析，结合色谱峰保留时间、精确相对分子质量、碎片离子、中性丢失等信息进行化合物指认。采用SwissTarget Predection预测药物靶点。基于GeneCards、OMIM、TTD、DisGeNET数据库获取疾病靶点，使用STRING数据库构建关键靶点蛋白相互作用（PPI）网络，利用Cytoscape软件构建核心靶点基因网络图和“成分-靶点-通路”图，通过Metascape数据库进行基因本体（GO）和京都基因和基因组百科全书（KEGG）通路富集分析。基于CB-DOCK2数据库对关键的活性成分和作用靶点进行分子对接验证，对分子对接后结合能小于−8 kcal/mol的配体与蛋白质复合物通过Gromacs 2022进行分子动力学模拟。结果 萆薢分清丸体外定性得到36个化学成分，大鼠ig给药后在血清样本中检测到18个入血成分。其中山柰酚、白杨素、乔松素等为萆薢分清丸发挥治疗慢性肾衰药效的关键成分，蛋白激酶B1（Akt1）、白蛋白（ALB）、表皮生长因子受体（EGFR）、信号转导及转录激活因子3（STAT3）、原癌基因酪氨酸蛋白激酶Src（SRC）等为萆薢分清丸治疗慢性肾衰的主要靶点，涉及到癌症通路、丝裂原活化蛋白激酶（MAPK）信号通路、叉头框转录因子O亚族信号（FoxO）信号通路等。分子对接显示Akt1、ALB、EGFR、STAT3、SRC靶点与药物成分具有较高的亲和力。分子动力学进一步验证大黄素与ALB、大黄素与EGFR、乔松素与ALB和山柰酚与STAT3靶点结合稳定。结论 萆薢分清丸可能通过山柰酚、大黄素、乔松素等关键药效成分，作用于Akt1、STAT3、EGFR等核心靶点，调控MAPK、FoxO、缺氧诱导因子‑1（HIF-1）等信号通路等发挥治疗慢性肾衰作用。

Objective To explores the potential active ingredients of Bixie Fenqing Pills, and the key targets for its treatment of chronic renal failure based on serum pharmacology, network pharmacology and molecular dynamics simulation. Methods In vitro and in vivo chemical profiling of Bixie Fenqing Pills was characterized using UHPLC-Q-Orbitrap HRMS. Compound identification was performed by integrating retention times, accurate molecular masses, fragment ions, and neutral loss patterns. Drug targets were predicted using SwissTargetPrediction. Disease-related targets were retrieved from GeneCards, OMIM, TTD, and DisGeNET databases. PPI networks of key targets were constructed using the STRING database. Cytoscape was utilized to visualize core target gene networks and “component–target–pathway” relationships. GO and KEGG pathway enrichment analyses were carried out through the Metascape platform. Molecular docking was performed to verify the binding affinities between key active compounds and core targets using CB-Dock2. Molecular dynamics simulations were conducted using GROMACS 2022 for ligand–protein complexes with docking energies below −8 kcal/mol. Results The external qualitative analysis of the Bixie Fenqing Pills yielded 36 chemical components. After intragastric administration to rats, 18 components that entered the bloodstream were detected in the serum samples. Among them, kaempferol, chrysin, and pinostrobin were the key components of Bixie Fenqing Pills that exert therapeutic effects on chronic renal failure. Akt1, ALB, EGFR, STAT3, and SRC were the main targets of Bixie Fenqing Pills in treatment of chronic renal failure, involving cancer pathways, MAPK signaling pathways, and FoxO signaling pathways, etc. Molecular docking shows that the Akt1, ALB, EGFR, STAT3, and SRC targets had a high affinity with the drug components. Molecular dynamics further validates the stable binding of emodin with ALB, emodin with EGFR, pinostrobin with ALB, and kaempferol with STAT3 targets. Conclusion Kaempferol, emodin, and pinostrobin are considered as the key bioactive components of Bixie Fenqing Pills, which may exert therapeutic effects against chronic renal failure by acting on the core targets (Akt1, STAT3, EGFR, etc.) and regulating critical pathways such as MAPK, FoxO, and HIF-1 signaling pathways.

R287.3

河南省高等学校重点科研项目（24A360025）