ObjectiveEpimedium (EPI) is a common Chinese herb with neuroprotective effects against a variety of central nervous system disorders, especially spinal cord injury (SCI). In this study, we performed network pharmacology and molecular docking analyses to reveal the mechanism underlying EPI treatment of SCI, then validated its efficacy using animal models.MethodsThe active ingredients and targets of EPI were screened by Traditional Chinese Medicine Systems Pharmacology (TCMSP) and their targets annotated on the UniProt platform. SCI-related targets were searched from OMIM, TTD, and GeneCards databases. We employed the STRING platform to construct a protein–protein interaction (PPI) network then visualized the results using Cytoscape (3.8.2) software. We also subjected key EPI targets to ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, then docked the main active ingredients with the key targets. Finally, we established an SCI rat model to evaluate efficacy of EPI in treating SCI and validate the effects of different biofunctional modules predicted by network pharmacology.ResultsA total of 133 EPI targets were associated with SCI. GO terms and KEGG pathway enrichment results showed that EPI’s effect in treating SCI was significantly associated with inflammatory response, oxidative stress and the PI3K/AKT signaling pathway. Molecular docking results indicated that EPI’s active ingredients have a high affinity for the key targets. Results from animal experiments revealed that EPI not only markedly improved Basso, Beattie, and Bresnahan scores in SCI rats, but also significantly improved p-PI3K/PI3K and p-AKT/AKT ratio. Moreover, EPI treatment not only mediated a significant decrease in malondialdehyde (MDA) but also increased both superoxide dismutase (SOD), and glutathione (GSH). However, this phenomenon was successfully reversed by LY294002, a PI3K inhibitor.ConclusionEPI improves behavioral performance in SCI rats through anti-oxidative stress, which may be mediated by activation of the PI3K/AKT signaling pathway.

Background: Spinal cord injury (SCI) is a type of trauma to the nervous system that can lead to severe dysfunction of the motor and sensory systems. The inhibitory microenvironment is the basis of secondary injury after SCI, and ferroptosis may aggravate secondary SCI. In this study, the role of ferroptosis inhibitor UAMC-3203 in the recovery of SCI was explored by examining the tissue structure, biochemical indexes, and motor function of rats after SCI. Methods: A model of SCI was established using Alice's method. The protective effect of UAMC-3203 on spinal cord neurons and the recovery of motor function were studied by Basso, Beattie, Bresnahan (BBB) score, incline plate test, Hematoxylin-eosin (HE) staining and Luxol Fast Blue (LFB) staining. Immunofluorescence and Elisa analysis were used to study the effect of UAMC-3203 on the microenvironment. Western blot analysis was used to measure the level of ferroptosis markers. Reactive oxygen species (ROS) and malondialdehyde (MDA) were detected to reflect the level of oxidation products. Results: We found that in the UAMC-3203 group, the loss and demyelination of neurons was reduced, the activation and proliferation of astrocytes and microglia and the secretion of related inflammatory cytokines was restrained, and motor function recovery was faster and better compared with those in the SCI group. Conclusions: UAMC-3203 has played an active role in the recovery process of SCI, providing evidence that post-SCI microenvironment imbalances are associated with ferroptosis.

Network pharmacology integrated with experimental validation to explore the therapeutic role and potential mechanism of Epimedium for spinal cord injury.

Icariin (ICA) is the main active component of the traditional Chinese medicine Epimedium, which has been confirmed to play a neuroprotective role in spinal cord injury (SCI) models. Astrocyte activation is a key link in “inflammatory storms” after SCI, and eventually form glial scar to inhibit neurological recovery. Studies have shown that icariin can effectively inhibit inflammatory response and promote the recovery of nerve function after SCI, but the specific mechanism is still unclear. This study applied network pharmacology and in vivo experimental verification to explore the molecular mechanism of ICA in the treatment of SCI. A total of 37 targets were screened out, mainly enriched in response to hypoxia, response to abiotic stimulus, regulation of cellular response to stress, and the TGF-beta signaling pathway etc. In the animal experiments, the Basso, Beattie and Bresnahan scores and Hematoxylin Eosin staining revealed that ICA could effectively improve the behavioral and histological recovery of SCI rats. Then ICA was confirmed to reduce astrocyte GFAP, Vimentin, and the expression of Ki-67 after SCI. Furthermore, ICA could inhibit the TGF-beta signaling pathway. ICA could also inhibit the expression of Yap protein. However, when Yap protein was activated, ICA inhibited the TGF-beta signaling pathway and reactive astrocyte activation was reversed. To sum up, the mechanism of ICA treatment for SCI may involve the inhibition of reactive astrocyte activation by inhibiting the expression of Yap protein and down-regulating the TGF-beta signaling pathway.