Discovery of Potential Mechanism of Zhenzhu Tongluo Wan for the Treatment of Peripheral Neuropathy Based on the Strategy of"Network Pharmacology,Mass Spectrometry Component Identification and Molecular Docking"
Objective This study aimed to systematically investigate the pharmacological substances and potential mechanisms of Zhenzhu Tongluo Wan in the treatment of peripheral nerve disorders.Methods Based on the TCMSP database and related literature,the active ingredients of Zhenzhu Tongluo Wan were determined.Target proteins for these active ingredients and those associated with peripheral neuropathy were predicted using the TCMSP,SwissTarget Prediction,GeneCard,OMIM,and DisGeNET databases.The intersection of these two sets of proteins was once considered a key target for the treatment of peripheral neuropathy.A protein-protein interaction(PPI)network of the intersection targets was constructed using the STRING database and analyzed using Cytoscape 3.9.1 software.Key targets obtained from the network were subjected to GO enrichment analysis and pathway annotation analysis using the DAVID database.The UHPLC-Q-Orbitrap HRMS technique was used to characterize the major chemical components of Zhenzhu Tongluo Wan.In addition,molecular docking,animal and cell experiments were performed to explore the mechanisms underlying the treatment of peripheral neuropathy with Zhenzhu Tongluo Wan.Results A total of 147 potential active ingredients and 333 intersection targets of Zhenzhu Tongluo Wan were screened.GO function and KEGG pathway analysis revealed that the major pathways involved include MAPK signaling pathway,NF-κB signaling pathway,PI3K-Akt signaling pathway,MAPK cascade and inflammatory response,among others.Based on high-resolution mass spectrometry analysis,33 major active ingredients were identified from Zhenzhu Tongluo Wan,focusing on those compounds with a Dgree value greater than the median.Molecular docking results further demonstrate good binding interactions between compounds such as luteolin,quercetin,tryptophan,arginine and kaempferol found inZhenzhu Tongluo Wan and key targets.Western blotting experiments showed that luteolin,quercetin and tryptophan could significantly inhibit the expression of MAPK8,P65 and TNF proteins in inflammatory cells.Animal experiments showed that Zhenzhu Tongluo Wan significantly improved hyperalgesia in rats with diabetic peripheral neuropathy,which may be related to the modulation of the expression of MAPK8,P65 and TNF proteins in the dorsal root ganglion.Conclusion Zhenzhu tongluo pills may play a role in the treatment of peripheral neuropathy by acting on the targets of MAPK8,P65 and TNF.This study lays a foundation for further research on the mechanism and pharmacodynamic substances of Zhenzhu Tongluo Wan in the treatment of peripheral neuropathy.
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