Network Pharmacology and Rat Caudal-disc Puncture Validation of Yi Qi Huo Xue Decoction For Lumbar Disc Degeneration

In this study, network pharmacology, molecular docking, molecular dynamics simulation, and experimental validation were used to investigate the pharmacological basis and potential molecular mechanisms of Yi Qi Huo Xue Decoction (YQHXD) in lumbar disc degeneration (LDD). A total of 678 active compound-related targets of YQHXD and 296 potential LDD-associated therapeutic targets were identified. Protein-protein interaction network analysis and topological analysis identified several candidate targets, including AKT1, IL6, TNF, BCL2, and STAT3. Enrichment analyses using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes indicated that the candidate targets were linked to functions including protein binding and ATP binding, with pathway enrichment that included PI3K-AKT signaling. Molecular docking predicted interactions between selected active compounds, such as quercetin, Tetraneurin A, and isorhamnetin, and the core targets. Molecular dynamics simulations further suggested that the lowest-energy complexes remained stable. Seventy male Sprague-Dawley rats were randomly assigned to the control, sham-operated, model, positive control, and low-, medium-, and high-dose YQHXD groups. After the establishment of a coccygeal intervertebral disc puncture model in the model, positive control, and YQHXD groups, rats were administered YQHXD or celecoxib by oral gavage for 8 weeks, whereas the control and sham-operated groups received normal saline. Histopathological evaluation showed that YQHXD treatment was associated with reduced intervertebral disc degeneration in this model. Enzyme-linked immunosorbent assay, polymerase chain reaction, and western blot analyses showed changes in the mRNA and protein expression levels of selected targets, including AKT1, BCL2, and STAT3. These findings suggest that YQHXD may exert preclinical protective effects in LDD through multi-component and multi-target regulation involving inflammation- and apoptosis-related mechanisms. This study provides preliminary preclinical evidence supporting further investigation of YQHXD for LDD.