Network pharmacology-based evaluation of Longzhong Xiaozhong mixture for treating spinal cord injuries in athletes: implications for recovery and fitness enhancement

Abstract

Background: The Longzhong Xiaozhong Zhitong Mixture has demonstrated significant therapeutic efficacy in the clinical management of spinal cord injury, particularly notable in the context of athletic injuries. Despite its clinical success, the precise pharmacological mechanisms and specific targets of this mixture remain undefined. Objective: This study aims to delineate the action mechanism of the Longzhong Xiaozhong Zhitong Mixture in treating spinal cord injuries among athletes through network pharmacology and validate these findings via animal models. Methods: Active components and their targets within the mixture were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the China National Knowledge Infrastructure (CNKI) database. SCI-related genes were sourced from databases such as GeneCards, OMIM, DrugBank, and PharmGKB. The intersection of drug targets with SCI-related genes provided the specific therapeutic targets for the mixture in treating spinal cord injuries. A compound-target regulatory network was constructed using Cytoscape_v3.9.0, while the STRING database facilitated the construction of protein-protein interaction (PPI) networks. Core genes were pinpointed via Cytoscape, with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses performed using R software. Cellular experiments utilized in vitro models of the hypoxia-reoxygenation-induced blood-spinal cord barrier in rats, employing techniques such as immunofluorescence, Western blot (WB), and reverse transcription-polymerase chain reaction (RT-PCR) to validate network pharmacology findings. The study focused on unraveling the molecular biology of the lncRNA MALAT1-ERK/p38MAPK-AQP4 signaling axis in the treatment of SCI. Results: The analysis identified 190 active ingredients and 249 corresponding targets, with 204 common targets found between the mixture's potential targets and SCI-related targets. Subsequent screening highlighted 14 core PPI network targets, including JUN, STAT3, AKT1, FOS, and MAPK3. GO and KEGG analyses implicated the mixture in processes such as chemical stress response and oxidative stress response, with significant pathways including the MAPK signaling pathway, apoptosis, and vascular endothelial growth factor signaling pathway. Conclusion: This investigation elucidates the pharmacological underpinnings of the Longzhong Xiaozhong Zhitong Mixture's efficacy in SCI treatment among athletes, detailing its active compounds, critical targets, and involved signaling pathways. Preliminary animal model validations offer robust evidence for further exploration of its molecular mechanisms, underscoring the mixture's potential in enhancing recovery and fitness in the athletic population following spinal cord injuries.