Cell death in spinal cord injury (SCI) requires de novo protein synthesis. Calpain inhibitor E-64-d provides neuroprotection in SCI lesion and penumbra.
Degradation of cytoskeletal proteins by calpain, a Ca(2+)-dependent cysteine protease, may promote neuronal apoptosis in the lesion and surrounding areas following spinal cord injury (SCI). Clinically relevant moderate (40 g-cm force) SCI in rats was induced at T12 by a standardized weight-drop method. Internucleosomal DNA fragmentation or apoptosis in the lesion was inhibited by 24-h treatment of SCI rats with cycloheximide (1 mg/kg), indicating a requirement for de novo protein synthesis in this process. To prove an involvement of calpain activity in mediation of apoptosis in SCI, we treated SCI rats with a cell-permeable calpain inhibitor E-64-d (1 mg/kg). Following 24-h treatment, a 5-cm-long spinal cord section centered at the lesion was collected, and divided equally into five segments (1 cm each) to determine calpain activity, as shown by degradation of the 68-kD neurofilament protein (NFP), and apoptosis as indicated by internucleosomal DNA fragmentation. Neurodegeneration propagated from the site of injury to neighboring rostral and caudal regions. Both calpain activity and apoptosis were readily detectable in the lesion, and moderately so in neighboring areas of untreated SCI rats, whereas these were almost undetectable in E-64-d-treated SCI rats, and absent in sham animals. Results indicate that apoptosis in the SCI lesion and penumbra is prominently associated with calpain activity and is inhibited by the calpain inhibitor E-64-d providing neuroprotective benefit.
Ray
S K
SK
Department of Neurology, Medical University of South Carolina, Charleston, South Carolina 29425, USA.
Matzelle
D D
DD
Wilford
G G
GG
Hogan
E L
EL
Banik
N L
NL
D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D002154:Calpain / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D003513:Cycloheximide / Q000627:therapeutic use; D005260:Female; D006023:Glycoproteins / Q000494:pharmacology* / Q000627:therapeutic use; D007930:Leucine / Q000031:analogs & derivatives* / Q000494:pharmacology* / Q000627:therapeutic use; D011480:Protease Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D011500:Protein Synthesis Inhibitors / Q000627:therapeutic use; D051381:Rats; D017207:Rats, Sprague-Dawley; D013119:Spinal Cord Injuries / Q000188:drug therapy / Q000378:metabolism*
D006023:Glycoproteins; D011480:Protease Inhibitors; D011500:Protein Synthesis Inhibitors; C039304:calpain inhibitors; D003513:Cycloheximide; D002154:Calpain; D007930:Leucine; C108192:aloxistatin