The demonstration of increased calpain activity, immunostaining, and expression at the gene (mRNA) and protein levels concomitant with ultrastructural degeneration and loss of axon and myelin proteins in lesioned cord have implicated a pivotal role for calpain in tissue destruction in spinal cord injury (SCI). Calpain, stimulated by free radicals, also mediates apoptotic cell death. These findings suggested that the use of calpain and lipid peroxidation drugs as therapeutic agents would protect cells and maintain the axon-myelin structural unit by preventing protein degradation. In order to examine this hypothesis, we treated SCI animals with calpain inhibitors (calpeptin) and/or methlprednisolone (MP), and antiinflammatory and free-radical inhibitor. SCI (40 g/cm) was induced by weight-drop, and 1 mg calpeptin or 165 mg MP/kg were given intravenously (i.v.) for 24 hours. Untreated injured animals receiving vehicle served as controls. Lesion 68-kDa and 200-kDa neurofilament proteins (NFPs) were analyzed by sodium dodecylsulfate polyarcylamide gel electrophoresis (SDS-PAGE) and chemiluminescence, and the extent of protein loss was quantitated. Loss of protein in the lesion of untreated cord amounted to 47% compared to sham control, while that for calpeptin- or MP-treated rats was 25-30%. Combination treatment with calpeptin and MP was slightly more effective in preventing NFP degradation, compared to either when used alone. Apopotic cell death in SCI as characterized by internucleosomal DNA fragmentation was also reduced following treatment with the inhibitors. The inhibition of cytoskeletal protein degradation suggests that calpain and free-radical inhibitors may rescue cells and preserve and maintain membrane structure by preventing protein breakdown, preserving motor function, and being neuroprotective.