Table6_Investigation of the Multi-Target Mechanism of Guanxin-Shutong Capsule in Cerebrovascular Diseases: A Systems Pharmacology and Experimental Assessment.XLSX

Guanxin-Shutong capsule (GXSTC), a combination of Mongolian medicines and traditional herbs, has been clinically proven to be effective in treating cerebrovascular diseases (CBVDs). However, the underlying pharmacological mechanisms of GXSTC in CBVDs remain largely unknown. In this study, a combination of systems pharmacology and experimental assessment approach was used to investigate the bioactive components, core targets, and possible mechanisms of GXSTC in the treatment of CBVDs. A total of 15 main components within GXSTC were identified using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD) and a literature research. Fifty-five common genes were obtained by matching 252 potential genes of GXSTC with 462 CBVD-related genes. Seven core components in GXSTC and 12 core genes of GXSTC on CBVDs were further determined using the protein-protein interaction (PPI) and component-target-pathway (C-T-P) network analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis results predicted that the molecular mechanisms of GXSTC on CBVDs were mainly associated with the regulation of the vascular endothelial function, inflammatory response, and neuronal apoptosis. Molecular docking results suggested that almost all of core component-targets have an excellent binding activity (affinity < −5 kcal/mol). More importantly, in middle cerebral artery occlusion (MCAO) -injured rats, GXSTC significantly improved the neurological function, reduced the infarct volume, and decreased the percentage of impaired neurons in a dose-dependent manner. Western blotting results indicated that GXSTC markedly upregulated the expression of vascular endothelial growth factor A (VEGFA) and endothelial nitric oxide synthase (eNOS), while downregulating the expression of cyclooxygenase-2 (COX-2) and transcription factor AP-1 (c-Jun) in MCAO-injured rats. These findings confirmed our prediction that GXSTC exerts a multi-target synergetic mechanism in CBVDs by maintaining vascular endothelial function, inhibiting neuronal apoptosis and inflammatory processes. The results of this study may provide a theoretical basis for GXSTC research and the clinical application of GXSTC in CBVDs.

关馨-舒通胶囊（GXSTC），由蒙古族药物及传统草药组成，经临床验证，对脑血管疾病（CBVDs）具有显著疗效。然而，GXSTC在CBVDs治疗中的潜在药理学机制尚不明确。本研究采用系统药理学与实验评估相结合的方法，探究GXSTC的生物活性成分、核心靶点和可能的药理作用机制。通过高效液相色谱联用二极管阵列检测器（HPLC-DAD）和文献研究，共鉴定出GXSTC中的15种主要成分。通过将GXSTC的252种潜在基因与462种CBVD相关基因进行匹配，获得55种常见基因。利用蛋白质-蛋白质相互作用（PPI）和成分-靶点-通路（C-T-P）网络分析，进一步确定了GXSTC中的7个核心成分和12个CBVDs相关的核心基因。基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析结果表明，GXSTC对CBVDs的分子机制主要与血管内皮功能调节、炎症反应和神经元凋亡有关。分子对接结果显示，几乎所有核心成分-靶点均表现出优异的亲和力（亲和力< -5 kcal/mol）。更重要的是，在中脑动脉闭塞（MCAO）损伤的大鼠模型中，GXSTC显著改善了神经功能，剂量依赖性地减少了梗死体积和受损神经元百分比。蛋白质印迹分析结果显示，GXSTC在MCAO损伤大鼠中显著上调了血管内皮生长因子A（VEGFA）和内皮型一氧化氮合酶（eNOS）的表达，同时下调了环氧合酶-2（COX-2）和转录因子AP-1（c-Jun）的表达。这些发现证实了我们的预测，即GXSTC通过维持血管内皮功能、抑制神经元凋亡和炎症过程，在CBVDs中发挥多靶点协同作用。本研究结果为GXSTC的研究及其在CBVDs临床应用提供了理论依据。