Mechanism of Qingre Huatan Tongfu formula in treating ischemic stroke based on network pharmacology

Objective: To explore the potential mechanism of Qingre Huatan Tongfu formula (HTF) in the treatment of ischemic stroke (IS) based on network pharmacology and animal experiments.Methods: The active ingredients of HTF were screened using TCMSP and BATMAN databases and the potential targets of these active ingredients were predicted.Disease-related targets were screened in GeneCards, OMIM, TTD, and Drugbank databases.The protein interaction relationship was obtained using the String database.Key targets underwent GO and KEGG enrichment analysis using Metascape.The animal model was constructed for experimental verification.Neural function scores, 2, 3, 5-triphenyltetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, Nissl staining, terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining and immunohistochemistry were used to verify the network pharmacological enrichment results.Results: By conducting network pharmacological analysis, the main active ingredients were predicted to include apigenin, quercetin, kaempferol, and β-sitosterol.These ingredients acted on potential targets such as serine/threonine kinase 1 (AKT1), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), transformation-associated protein 53(TP53)and other potential targets.The key pathways involved included lipid and atherosclerosis, MAPK signaling pathway, PI3K-Akt signaling pathway, TNF signaling pathway, IL-17 signaling pathway, and hypoxia-inducible factor-1 (HIF-1)signaling pathway, etc.The experimental results showed that HTF could significantly reduce the nerve function score and the volume of cerebral infarction in cerebral ischemia-reperfusion rats (P< 0.05), improve the survival of neurons(P< 0.05), and significantly reduce the cell apoptosis(P< 0.05).Conclusion: Based on network pharmacology and animal experiments, it is found that HTF can play a role in the treatment of IS through multiple components acting on multiple targets, involving various pathways such as anti-inflammation and anti-apoptosis.