Synergistic effects of caspase inhibitors and MK-801 in brain injury after transient focal cerebral ischaemia in mice

Abstract

1. Excitotoxic and apoptotic mechanisms have been implicated in the pathophysiology of cerebral ischaemia. Both MK-801, an NMDA receptor antagonist, or peptide inhibitors of the caspase family (z-VAD.FMK and z-DEVD.FMK), protect mouse brain from ischaemic cell damage. In this study, we examined whether these drugs which act via distinct mechanisms, afford even greater neuroprotection when given in combination following 2 h MCA occlusion (filament model) and 18 h reperfusion.

2. Given alone as pretreatment, MK-801 (1, 3 and 5 mg kg⁻¹, but not 0.3 mg kg⁻¹, i.p.) decreased infarct size by 34–75%. When injected 1 h after occlusion and before reperfusion, 3 mg kg⁻¹ reduced injury but not when administered 1 h after reperfusion.

3. Pretreatment with a subthreshold dose of MK-801 (0.3 mg kg⁻¹) plus a subthreshold dose of z-VAD.FMK (27 ng) or z-DEVD (80 ng) significantly decreased infarct size by 29 and 30%, respectively, and enhanced neurological function.

4. Administering a subthreshold dose of z-VAD.FMK (27 ng) or z-DEVD.FMK (80 ng) as pretreatment extended the time window for MK-801 (3 mg kg⁻¹) by 2 h from 1 h before reperfusion to at least 1 h after reperfusion.

5. Pretreating with a subthreshold dose of MK-801 (0.3 mg kg⁻¹) extended the time window for z-DEVD.FMK (480 ng) from 1 h after reperfusion to at least 3 h after reperfusion.

6. We conclude that caspase inhibitors which putatively block apoptotic cell death and inhibit cytokine production and the NMDA antagonist MK-801 act synergistically and prolong their respective therapeutic windows in cerebral ischaemia.