Sheng-ji Hua-yu Formula promotes diabetic ulcer healing via regulating the cAMP/PKA/CREB signaling pathway.

Ethnopharmacological Relevance: Diabetic Ulcers (DUs) represent a common and severely debilitating complication associated with diabetes. Chinese Herbal Medicine (CHM) provides advantages in the treatment and healing of DUs. Sheng-ji Hua-yu Formula (SJHY), a topical CHM prescription with a long history of clinical application, has been proven to effectively accelerate healing processes in patients with DUs and exhibits a definite therapeutic effect while ensuring safety. However, the specific mechanisms underlying the SJHY treatment require further investigation.

Aim Of The Study: The purpose of this study was to investigate the underlying mechanisms through which SJHY can be utilized in the treatment of DUs.

Materials And Methods: Characterization of the chemical constituents of SJHY were analyzed using Liquid chromatography-tandem mass spectrometry (LC-MS/MS). Based on the identified targets, we then conducted a network pharmacology study, the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment and Gene Ontology (GO) analyses for compound-target prediction and analysis. Following treatment with SJHY, RNA-seq analysis was performed to investigate differentially expressed genes (DEGs). Using network pharmacology, protein-protein interaction (PPI) network, KEGG and GO, we explored the core signaling pathways and targets. These findings were subsequently validated through immunoblotting and immunofluorescence. Lipinski's Rule of Five and molecular docking were used to identify the compounds that interacted with the core pathways. The interaction between the core receptors and SJHY components was further investigated using molecular dynamics simulations.

Results: A total of 202 compounds in SJHY and 2881 specific targets for these compounds were discovered. Analysis using KEGG and GO highlighted the potential importance of the cAMP pathway in the therapeutic effects of SJHY. Between the SJHY-treated group and the model group, there were 476 DEGs, with 311 genes being up-regulated and 165 genes being down-regulated. Network pharmacology, PPI networks, KEGG pathways and GO annotations, all indicated that the cAMP signaling pathway was a critical route through which SJHY exerts its effects on DUs. By applying Lipinski's Rule of Five and molecular docking, four active compounds in SJHY that interacted with the core pathways were identified. Molecular dynamics simulations (MDs) further demonstrated a robust binding affinity between the SJHY components and the core target.

Conclusion: Our findings offer an in-depth insight into the treatment of DUs in herbal intervention. By enriched the multifaceted components and numerous targets of SJHY, it suggests that a focused approach on the cAMP signaling pathway could represent a groundbreaking therapeutic solution for treating DUs using CHM.