The effect of ischemic post-conditioning on hippocampal cell apoptosis following global brain ischemia in rats.

We evaluated the effect of brain ischemic post-conditioning on cell apoptosis in the hippocampus following global brain ischemia in rats. Adult male Sprague-Dawley rats were randomly divided into three groups (n=15/group): sham operation, ischemia/reperfusion (I/R) and ischemic post-conditioning (I PostC). Global brain ischemia was induced by four-vessel occlusion. Ischemic post-conditioning consisted of six cycles of 10s/10s reperfusion/reocclusion at the onset of reperfusion. All rats were sacrificed 24 hours or 72 hours after reperfusion. The hippocampal CA1 regions were analysed using the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labelling (Tunel) staining technique for determining cell apoptosis. Levels of caspase-3 and Bcl-2 were measured by Western blotting. After 72 hours, fewer Tunel-positive brain cells were observed in rats from the I PostC group than in rats from the I/R group (10.3 ± 2.7% versus 40.8 ± 6.2%, p<0.01). After reperfusion at 24 hours and 72 hours, expression of caspase-3 in the I PostC group was significantly decreased (p<0.01) and expression of Bcl-2 in the I PostC group was significantly increased (p<0.01) compared with the I/R group. We conclude that down-regulation of caspase-3 and up-regulation of Bcl-2 by ischemic post-conditioning may underlie the protective effects of post-conditioning.

Neuroprotective effects of ischemic postconditioning on global brain ischemia in rats through upregulation of hippocampal glutamine synthetase.

Brain ischemic postconditioning is the induction of brief periods of ischemia-reperfusion during the early stages following ischemia, and it has been shown to produce neuroprotective effects. The mechanisms underlying these neuroprotective effects are poorly understood. Glutamate excitotoxicity is one cause of postischemic neuronal death. Glutamine synthetase (GS) is an enzyme that is expressed in glial cells and may affect glutamate excitotoxicity. We induced global ischemia in rats and performed postconditioning with 6 cycles of 10 seconds reperfusion and 10 seconds reocclusion before final reperfusion. Hematoxylin and eosin staining revealed extensive neuronal loss (44.0 ± 2.8% cell survival) in the hippocampal CA1 region. Ischemic postconditioning decreased neuronal death (82.0 ± 5.6% cell survival; p<0.05). Western blotting revealed significantly increased GS expression in the hippocampus for the ischemia-reperfusion group over time compared with the sham group (p<0.05). Ischemic postconditioning resulted in significantly increased (p<0.05) GS expression compared with both the sham and ischemia-reperfusion groups, suggesting that upregulation of GS expression after ischemia constitutes a neuroprotective mechanism.

Involvement of Glutamate transporter-1 in neuroprotection against global brain ischemia-reperfusion injury induced by postconditioning in rats.

Ischemic postconditioning refers to several transient reperfusion and ischemia cycles after an ischemic event and before a long duration of reperfusion. The procedure produces neuroprotective effects. The mechanisms underlying these neuroprotective effects are poorly understood. In this study, we found that most neurons in the CA1 region died after 10 minutes of ischemia and is followed by 72 hours of reperfusion. However, brain ischemic postconditioning (six cycles of 10 s/10 s reperfusion/re-occlusion) significantly reduced neuronal death. Significant up-regulation of Glutamate transporter-1 was found after 3, 6, 24, 72 hours of reperfusion. The present study showed that ischemic postconditioning decreases cell death and that upregulation of GLT-1 expression may play an important role on this effect.