Melatonin improves functional outcome via inhibition of matrix metalloproteinases-9 after photothrombotic spinal cord injury in rats.

Abstract

Matrix metalloproteinases (MMPs), especially MMP-2 and MMP-9 play an important role in secondary inflammatory reaction and blood-central nervous system (CNS) barrier disruption after spinal cord injury (SCI). Theoretically, it is expected that early blockade of activation of MMPs can provide neuro-protective effects from secondary tissue damage and improve functional neurological outcomes. The aim of this study was to investigate the expression and the activity of MMP-2 and MMP-9, and to determine the regulatory effect of melatonin on MMP expression and activity after photochemically induced SCI in rats. Female Sprague-Dawley rats weighing between 250 and 300 g (age 8 weeks) received focal ischemia by photothrombosis using Rose Bengal (RB). The injured animals were divided into two groups; one group received 50 mg/kg of melatonin intraperitoneally, starting 1 h after injury and at 12 h intervals for 7 days, while animals in the control group received weight-adjusted doses of a saline vehicle. In each group, the expressions and activities of MMP-2 and MMP-9 were assessed by Western blot and gelatin zymography at various times from 6 h to 3 days. The locomotor function was assessed using the Basso-Beattie-Bresnahan (BBB) scale at 3 days after SCI and then once per week for 4 weeks. The animals were killed at 28 days after the injury, and the histopathology of the lesions was assessed. The expressions and activities of MMP-9 were increased at 6, 24, 48, and 72 h after SCI in the control group. In the melatonin-treated group, the expression of MMP-9 was significantly decreased at 24, 48, and 72 h after SCI compared with the control group, and the activity of MMP-9 was significantly reduced at 72 h after SCI. In contrast, there were no significant changes in the MMP-2 level in both groups during the experimental period. Melatonin treatment following photochemically induced SCI in rats significantly ameliorated the functional deficits. On histopathologic examination, the lesion size in the spinal cord after photothrombotic insult was significantly reduced by melatonin administration. This study showed that the up-regulation of MMP-9 correlated with the secondary damage after SCI in rats. The results of this study suggest that the ability of melatonin to reduce secondary tissue damage is intimately related to the reduction of MMP-9 expression, resulting in functional improvement.