Proteolytic enzymes in experimental spinal cord injury.
Experimental spinal cord injury was produced in rats by dropping a 10 g weight from 30 cm upon dura-invested exposed spinal cord. Proteolytic activities at neutral (pH 7.6) and acid (pH 5.5 and 3.6) pH were determined in whole homogenate and the cytosolic fraction of the lesion (lumbar) and cervical control segments. The enzyme activity was monitored by SDS-PAGE analysis of the extent of substrate myelin basic protein (MBP) degradation. Activities (neutral and cathepsin B-like) in the sham-operated spinal cord were lower than those of cervical autologous control at 24 h after injury. The increase in neutral proteinase activity was progressive and greater in the lesion than the autologous control. A 61.5% +/- 3.5 loss of MBP was observed at 2 h following injury and increased at 24 h (78.2% +/- 3.4). The loss of MBP coincided with the appearance of several low molecular weight peptides. The cathepsin B-like and cathepsin D activities were also increased in the lesion but to a lesser extent than the neutral proteinase. The neutral proteinase and cathepsin B-like activity were inhibited by leupeptin and not by pepstatin while the converse obtained for cathepsin D activity. The release of neutral proteolytic activity which is nonlysosomal in origin suggests a novel hypothesis for the mechanism of traumatic axon-myelin injury.
D000818:Animals; D002384:Catalysis; D002401:Cathepsin B; D002402:Cathepsin D / Q000378:metabolism; D002403:Cathepsins / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D004676:Myelin Basic Protein / Q000378:metabolism; D051381:Rats; D013116:Spinal Cord / Q000201:enzymology*; D013119:Spinal Cord Injuries / Q000201:enzymology*
D004676:Myelin Basic Protein; D002403:Cathepsins; D010450:Endopeptidases; D002401:Cathepsin B; D002402:Cathepsin D