RESUMEN
Salvianolic acid A (Sal A), a bioactive compound isolated from the Chinese medicinal herb Danshen, is used for the prevention and treatment of cardiovascular diseases. However, the protective function of Sal A on preserving the role of blood-spinal cord barrier (BSCB) after spinal cord injury (SCI) is unclear. The present study investigated the effects and mechanisms of Sal A (2.5, 5, 10mg/kg, i.p.) on BSCB permeability at different time-points after compressive SCI in rats. Compared to the SCI group, treatment with Sal A decreased the content of the Evans blue in the spinal cord tissue at 24h post-SCI. The expression levels of tight junction proteins and HO-1 were remarkably increased, and that of p-caveolin-1 protein was greatly decreased after SCI Sal A. The effect of Sal A on the expression level of ZO-1, occluding, and p-caveolin-1 after SCI was blocked by the HO-1 inhibitor, zinc protoporphyrin IX (ZnPP). Also, Sal A inhibited the level of apoptosis-related proteins and improved the motor function until 21days after SCI. In addition, Sal A significantly increased the expression of microRNA-101 (miR-101) in the RBMECs under hypoxia. AntagomiR-101 markedly increased the RBMECs permeability and the expression of the Cul3 protein by targeting with 3'-UTR of its mRNA. The expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and HO-1 was significantly increased after agomiR-101 treatment. Therefore, Sal A could improve the recovery of neurological function after SCI, which could be correlated with the repair of BSCB integrity by the miR-101/Cul3/Nrf2/HO-1 signaling pathway.
Asunto(s)
Ácidos Cafeicos/farmacología , Permeabilidad Capilar/efectos de los fármacos , Fármacos Cardiovasculares/farmacología , Lactatos/farmacología , Traumatismos de la Médula Espinal/tratamiento farmacológico , Médula Espinal/irrigación sanguínea , Médula Espinal/efectos de los fármacos , Animales , Permeabilidad Capilar/fisiología , Caveolina 1/metabolismo , Proteínas Cullin/metabolismo , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Hemo Oxigenasa (Desciclizante)/metabolismo , Hipoxia/tratamiento farmacológico , Hipoxia/metabolismo , Masculino , MicroARNs/metabolismo , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , Factor 2 Relacionado con NF-E2/metabolismo , Fármacos Neuroprotectores/farmacología , Distribución Aleatoria , Ratas Sprague-Dawley , Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/metabolismo , Uniones Estrechas/efectos de los fármacos , Uniones Estrechas/metabolismo , Proteína de la Zonula Occludens-1/metabolismoRESUMEN
This study was performed to investigate the effect of bone marrow stromal cells (BMSCs) combined with green tea polyphenols (GTPs) on the blood-spinal cord barrier (BSCB) permeability after spinal cord injury (SCI) in the rat model. In the model of SCI rats, we found that the water content and the BSCB permeability were decreased by BMSCs and GTPs treatment, and their combination had a synergistic effect. Further, the motor function of rats was also greatly improved by BMSCs and GTPs administration. After treated by the combination of BMSCs and GTPs, SCI rats showed the up-regulated expression of tight junction (TJ) associated proteins claudin-5, occludin and ZO-1 by Western blot, which was more remarkable than that in the single treatment. The increased expression levels of claudin-5, occludin, and ZO-1 were the most obvious in the spinal cord microvessels using immunohistochemistry assay. This led to the conclusion that the combination of BMSCs and GTPs could decrease the BSCB permeability by up-regulating protein expression levels of claudin-5, occludin, and ZO-1. In addition, after BMSCs and GTPs administration, the results of Western blot and enzyme-linked immunosorbent assay (ELISA) revealed a significant decrease in protein expression level and the activation of nuclear factor-кB (NF-кB) p65. Our results indicated that combination of BMSCs and GTPs could improve motor function after SCI, which might be correlated with improvements in BSCB integrity, and that NF-кB might be involved in the modulating process.
Asunto(s)
Permeabilidad Capilar , Trasplante de Células Madre Mesenquimatosas , Extractos Vegetales/farmacología , Polifenoles/farmacología , Compresión de la Médula Espinal/terapia , Médula Espinal/irrigación sanguínea , Animales , Células Cultivadas , Claudina-1/genética , Claudina-1/metabolismo , Masculino , FN-kappa B/metabolismo , Ocludina/genética , Ocludina/metabolismo , Extractos Vegetales/uso terapéutico , Polifenoles/uso terapéutico , Ratas , Ratas Sprague-Dawley , Médula Espinal/efectos de los fármacos , Médula Espinal/metabolismo , Compresión de la Médula Espinal/tratamiento farmacológico , Té/química , Regulación hacia Arriba , Proteína de la Zonula Occludens-1/genética , Proteína de la Zonula Occludens-1/metabolismoRESUMEN
Responsible for orchestrating cellular energy production, mitochondria are central to the maintenance of life and the gatekeepers of cell death. Its morphology is dynamic and controlled by continual and balanced fission and fusion events. In this study, we analyzed the mitochondrial dynamics and functions after spinal cord injury in rats and further to discuss the mechanisms of the mitochondria regulated cell injury during SCI. Using adult rat spinal cord injury model, it was found that the absolute number of mitochondria per area was significantly less and the individual mitochondrial cross-sectional area was significantly greater in the neurons of rats in SCI group than in the sham-operated group at 3h and 6h after SCI, and the reverse pattern at 12h and 24h after SCI. The results from Western blot and RT-PCR assays showed that the protein and mRNA levels of mitochondrial fusion-related genes (Mfn1 and Mfn2) decreased and fission-related genes (Drp1 and Fis1) increased at 3h and 6h after SCI. At 12h and 24h after SCI the reverse pattern of Mfn1, Mfn2, Drp1 and Fis1 expression was found. Taken together the results of the present study showed the mitochondrial tendency of elongation and fusion in the injured spinal cord at 3h and 6h after SCI, and the tendency of mitochondrial fission at 12h and 24h after SCI in our SCI models of rat. These findings have important implications for our understanding of the mechanisms of mitochondrial dynamics and functions after SCI injury. And mitochondrial fusion may potentially be used as a target for improving spinal cord function in the first 6h after SCI. Mitochondrial fusion may be inhibited at 12-24h after SCI for improving functional outcomes following SCI.
Asunto(s)
Mitocondrias/patología , Dinámicas Mitocondriales/fisiología , Traumatismos de la Médula Espinal/patología , Traumatismos de la Médula Espinal/fisiopatología , Animales , Western Blotting , Masculino , Microscopía Electrónica de Transmisión , Proteínas Mitocondriales/análisis , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The current study was performed to investigate the effect of baicalin (BC) on spinal cord injury (SCI) in rat. BC (10, 30 and 100mg/kg, i.p., respectively) was administered to rats immediately and every 24h following SCI. The BC therapy (100mg/kg) dramatically decreased (1) the water content of spinal cord tissue (by dry-wet weight method), (2) the permeability of blood-spinal cord barrier (measured by Evans blue), (3) oxidant stress (malondialdehyde values and glutathione levels evaluation), (4) proinflammatory cytokines expression (tumor necrosis factor-α and NF-κB) (5) and apoptosis (measured by Bax, Bcl-2 and Caspase-3 expression). And the treatment with BC also significantly improved the recovery of limb function (evaluated by motor recovery score). Taken together, our results clearly indicate that BC possesses potent anti-inflammatory and anti-apoptotic properties, attenuates the SCI and is a new promising therapeutic agent for human SCI in the future.