【摘要】 目的：基于网络药理学和实验验证相结合的方法探讨丹参-红花药对协同心肌保护的作用机制。方法：通过网络药理学构建丹参-红花药对“活性成分-作用靶点-疾病”蛋白网络探讨其调控规律；借助分子对接技术验证成分与潜在靶标的结合能力；采用H9c2糖氧剥夺细胞模型评价活性成分的药效，结合联合指数（combination index, CI）评价成分间的协同效应；运用western blot检测靶标蛋白的表达。结果：网络药理学分析表明丹参-红花含有5条核心通路和8个核心靶点发挥心肌保护作用，其中5个核心靶点富集于HIF-1信号通路；丹参-红花主要通过调控HIF-1信号通路的靶基因进而调节血管张力；活性成分杨梅素、丹酚酸B、隐丹参酮能协同激活HIF-1通路的上游基因STAT3磷酸化。细胞实验结果表明，杨梅素、丹酚酸B、隐丹参酮具有显著的协同心肌保护作用。分子对接验证了杨梅素、丹酚酸B、隐丹参酮与STAT3靶点的强结合力。Western blot进一步表明，活性成分协同上调STAT3蛋白的表达。结论：药效学分析表明丹参-红花活性成分通过多靶点、多途径协同发挥心肌保护作用。本研究为阐释中药配伍的内涵提供方法学参考。更多还原

【Abstract】 Objective: To reveal the molecular mechanism underlying the compatibility of Salvia miltiorrhiza Bge(S. miltiorrhiza, Dan Shen) and C. tinctorius L.(C. tinctorius, Hong Hua) as an herb pair through network pharmacology and subsequent experimental validation.Methods: Network pharmacology was applied to construct an active ingredient-efficacy target-disease protein network to reveal the unique regulation pattern of S. miltiorrhiza and C. tinctorius as herb pair.Molecular docking was used to verify the binding of the components of these herbs and their potential targets. An H9c2 glucose hypoxia model was used to evaluate the efficacy of the components and their synergistic effects, which were evaluated using the combination index. Western blot was performed to detect the protein expression of these targets.Results: Network pharmacology analysis revealed 5 pathways and 8 core targets of S. miltiorrhiza and C. tinctorius in myocardial protection. Five of the core targets were enriched in the hypoxia-inducible factor-1(HIF-1) signaling pathway. S. miltiorrhiza-C. tinctorius achieved vascular tone mainly by regulating the target genes of the HIF-1 pathway. As an upstream gene of the HIF-1 pathway, STAT3 can be activated by the active ingredients cryptotanshinone(Ctan), salvianolic acid B(Sal. B), and myricetin(Myric). Cell experiments revealed that Myric, Sal. B, and Ctan also exhibited synergistic myocardial protective activity. Molecular docking verified the strong binding of Myric, Sal. B, and Ctan to STAT3.Western blot further showed that the active ingredients synergistically upregulated the protein expression of STAT3.Conclusion: The pharmacodynamic transmission analysis revealed that the active ingredients of S. miltiorrhiza and C. tinctorius can synergistically resist ischemia through various targets and pathways.This study provides a methodological reference for interpreting traditional Chinese medicine compatibility.更多还原