Neuroprotection at the initial stage of spinal cord injury (SCI) is one main goal to inhibit delayed network damage triggered by excitotoxicity mediated via hyperactivation of glutamatergic systems. Previous studies have shown positive effects of general inhalation anesthetics, while early neurosurgical operations can attenuate late pathophysiological events. It is interesting to find out if the widely used i.v. anesthetic propofol could protect locomotor spinal networks from excitotoxic insult induced by transient application of kainate (0.1 mM) for 1 h to the rat spinal cord in vitro. This protocol was designed to mimic the clinical onset of SCI followed by intensive care management. Significant neuronal losses were elicited by kainate, with almost halving of motoneuron numbers. Bath-applied propofol (5 µM, 4-8 h) potentiated GABA and depressed NMDA receptor responses together with decreased polysynaptic reflex activity which partially recovered after 24 h. Fictive locomotion evoked by repeated dorsal root stimuli or NMDA and serotonin (5HT) was weaker; however, when applied after kainate, there was no additive depression on synaptic transmission, suggesting that any further deterioration had been arrested. In all spinal areas and especially in the motoneuron pools, significant neuroprotection was observed after propofol administration following kainate. The periodicity of disinhibited bursts was improved by propofol after the kainate insult in line with the good histological preservation. These results suggest propofol could exert a delayed neuroprotective action on spinal network excitotoxic damage in an in vitro SCI model.