Increased muscle proteolysis after local trauma mainly reflects macrophage-associated lysosomal proteolysis.
Rat gastrocnemius showed increased protein degradation (+75-115%) at 48 h after traumatic injury. Injured muscle showed increased cathepsin B activity (+327%) and mRNA encoding cathepsin B (+670%), cathepsin L (+298%), cathepsin H (+159%), and cathepsin C (+268%). In in situ hybridization, cathepsin B mRNA localized to the mononuclear cell infiltrate in injured muscle, and only background levels of hybridization were observed either over muscle cells in injured tissue or in uninjured muscle. Immunogold/electron microscopy showed specific staining for cathepsin B only in lysosome-like structures in cells of the mononuclear cell infiltrate in injured muscle. Muscle cells were uniformly negative in the immunocytochemistry. Matrix metalloproteinase-9 (granulocyte-macrophage gelatinase) mRNA and activity were not present in uninjured muscle but were expressed after trauma. There was no activation of the ATP-ubiquitin-proteasome-dependent proteolytic pathway in injured muscle, by contrast to diverse forms of muscle wasting where the activity of this system and the expression of genes encoding ubiquitin and proteasome elements rise. These results suggest that proteolytic systems of the muscle cells remain unstimulated after local injury and that lysosomal enzymes of the inflammatory infiltrated cells are likely to be the major participant in protein catabolism associated with local trauma.
Farges
Marie-Chantal
MC
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, T6G 2P5, Canada.
Balcerzak
Denis
D
Fisher
Brian D
BD
Attaix
Didier
D
Béchet
Daniel
D
Ferrara
Marc
M
Baracos
Vickie E
VE
D000818:Animals; D003288:Contusions / Q000378:metabolism* / Q000473:pathology; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism; D018093:Gelatinases / Q000378:metabolism; D015870:Gene Expression; D017403:In Situ Hybridization; D066298:In Vitro Techniques; D008247:Lysosomes / Q000201:enzymology*; D008264:Macrophages / Q000201:enzymology*; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D018482:Muscle, Skeletal / Q000293:injuries* / Q000378:metabolism* / Q000473:pathology; D010447:Peptide Hydrolases / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D012333:RNA, Messenger; D010450:Endopeptidases; D010447:Peptide Hydrolases; D018093:Gelatinases; D020780:Matrix Metalloproteinase 9