Abstract
Background: Spinal cord injury (SCI) is a severe central nervous system injury for which few efficacious drugs are available. Rosmarinic acid (RA), a water-soluble polyphenolic phytochemical, has antioxidant, anti-inflammatory, and anti-apoptotic properties. However, the effect of RA on SCI is unclear. We investigated the therapeutic effect and underlying mechanism of RA on SCI in vivo and in vitro. Methods: In vivo experiment, The BBB locomotion scale, the inclined plane test, Nissl staining, and spinal cord edema were employed to determine the neuroprotective effects of RA treatment after SCI. Inflammatory and oxidative stress markers were detected by commercial kits and cell apoptosis status was measured by TUNEL staining. A proteomics and bioinformatics approach, together with Western blotting, was used to investigate the effect of RA on the proteome of SCI rats. In vitro experiment, oxidative stress and inflammatory injury were induced by H2O2 and LPS stimulation. Effects of RA on cell viability, apoptosis, inflammatory, and oxidative stress were evaluated. Results: Using a rat model of SCI, we showed that RA improved locomotor recovery after SCI and significantly mitigated neurological deficit, increased neuronal preservation, and reduced apoptosis. Also, RA inhibited activation of microglia and the release of TNF-α, IL-6, and IL-1β and MDA. Moreover, proteomics analyses identified the Nrf2 and NF-κB pathways as targets of RA. Pretreatment with RA increased levels of Nrf2 and HO-1 and reduced those of TLR4 and MyD88 as well as phosphorylation of IkB and subsequent nuclear translocation of NF-κB-p65. Using H2O2- and LPS-induced PC12 cells, we found that RA ameliorated the H2O2-induced decrease in viability and increase in apoptosis and oxidative injury by activating the Nrf2/HO-1 pathway. Also, LPS-induced cytotoxicity and increased apoptosis and inflammatory injury in PC-12 cells were mitigated by RA by inhibiting the TLR4/NF-κB pathway. The Nrf2 inhibitor ML385 weakened the effect of RA on oxidant stress, inflammation and apoptosis in SCI rats, and significantly increased the nuclear translocation of NF-κB. Conclusions: Therefore, the neuroprotective effect on SCI of RA may be due to its antioxidant and anti-inflammatory properties, which are mediated by modulation of the Nrf2/HO-1 and TLR4/NF-κB pathways. Moreover, RA activated Nrf2/HO-1, which amplified its inhibition of the NF-κB pathway.
MP Resulting in Autophagic Cell Death of Microglia through Zinc Changes against Spinal Cord Injury
