Proteasome alteration and delayed neuronal death in hippocampal CA1 and dentate gyrus regions following transient cerebral ischemia

BACKGROUND:Proteasome dysfunction has been reported to induce abnormal protein aggregation and cell death. OBJECTIVE:To investigate the effect of proteasome changes on delayed neuronal death in CA1 and dentate gyrus(DG) regions of the rat hippocampus following transient cerebral ischemia. DESIGN,TIME AND SETTING:A randomized,controlled animal experiment.The study was performed at the Department of Biochemistry and Molecular Biology,Norman Bethune Medical College of Jilin University,from September 2006 to May 2008. MATERIALS:Rabbit anti-19S S10B polyclonal antibody was purchased from Bioreagents,USA; propidium iodide and fluorescently-labeled goat anti-rabbit IgG were purchased from Jackson Immunoresearch,USA;hematoxylin and eosin staining solution was purchased from Sigma,USA; LSM 510 confocal microscope was purchased from Zeiss,Germany. METHODS:A total of 40 healthy Wistar rats,male,4 months old,were randomly divided into sham surgery group(n = 8) and model group(n = 32).Ischemic models were established in the model group by transient clamping of the bilateral carotid arteries and decreased blood pressure. After 20 minutes of global ischemia,the clamp was removed to allow blood flow for 30 minutes,4,24, and 72 hours,respectively,with 8 rats at each time point.The bilateral carotid arteries were not ligated in the sham surgery group. MAIN OUTCOME MEASURES:Neuronal death in the CA1 and DG regions was observed by hematoxylin-eosin staining.Proteasome expression in CA1 and DG region neurons was detected by immunohistochemistry. RESULTS:Hematoxylin-eosin staining showed neuronal death in the CA1 region alone at 72 hours of reperfusion following ischemia.In comparison to the sham surgery group,a significant decrease in proteasome expression was observed,by immunohistochemistry,in the CA1 and DG regions in the model group,following 30 minutes,4,24,and 72 hours of reperfusion(P<0.01).After 72 hours of reperfusion following ischemia,proteasome expression had almost completely disappeared in the CA1 region.In contrast,neurons of the DG region showed minimized proteasome expression at 24 hours,with a slight increase at 72 hours(P<0.01). CONCLUSION:The alteration of proteasome following ischemia/reperfusion in the neurons of hippocampal CA1 and DG regions reduces the ability of cells to degrade abnormal protein,which may be an important factor resulting in delayed neuronal death following transient cerebral ischemia.