Transgenic expression of human complement regulators reduces skeletal muscle ischaemia/reperfusion injury in mice.
This study aimed to explore the hypothesis that activated complement components contribute significantly to I/R (ischaemia/reperfusion) injury in skeletal muscle. After 50, 70 and 90 min of tourniquet ischaemia and 24 h of reperfusion, viability of the medial gastrocnemius muscle in CBA-C57BL/6 wild-type mice, assessed histochemically by reduction of NBT (Nitro Blue Tetrazolium) dye, was 60, 21 and 8% respectively. Skeletal muscle viability after 70 min of ischaemia and 24 h of reperfusion in transgenic mice expressing a combination of human CD46, CD55 and CD59, all inhibitors of complement activation, was 45% compared with 24% in ischaemic reperfused wild-type mice (P=0.008; n=6 per group). Muscle from sham-treated transgenic mice and wild-type littermates had no significant loss of viability relative to normal contralateral gastrocnemius muscle. A significant reduction in myeloperoxidase activity (a measure of neutrophil infiltration), xanthine oxidase activity (a source of free radicals) and water content (a measure of oedema) was observed in ischaemic reperfused muscle from transgenic mice compared with ischaemic reperfused wild-type muscle (P<0.05). Haematoxylin and eosin-stained histological sections also showed less damage and less apparent leucocyte infiltration in muscles from ischaemic reperfused transgenic mice than those from wild-type animals given the same degree of injury. Muscles from sham-treated transgenic and wild-type controls were almost identical with normal muscle. It is concluded that complement activation contributes to the pathogenesis of I/R injury in murine skeletal muscle, resulting in increased neutrophil infiltration into the injured muscle, increased free radical production and vascular permeability during reperfusion, and a net detrimental effect on muscle viability.
Knight
Kenneth R
KR
Bernard O'Brien Institute of Microsurgery, St. Vincent's Hospital Melbourne, Fitzroy, Victoria 3065, Australia. knightkr@svhm.org.au
Shinkel
Trixie A
TA
Cowan
Peter J
PJ
Romeo-Meeuw
Rosalind
R
d'Apice
Anthony J F
AJ
Morrison
Wayne A
WA
D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D018958:CD55 Antigens / Q000235:genetics; D018957:CD59 Antigens / Q000235:genetics; D003169:Complement Inactivator Proteins / Q000235:genetics* / Q000276:immunology; D004487:Edema / Q000378:metabolism; D006614:Hindlimb; D006801:Humans; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D023421:Models, Animal; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000201:enzymology / Q000276:immunology; D020556:Neutrophil Infiltration; D009195:Peroxidase / Q000032:analysis; D015427:Reperfusion Injury / Q000201:enzymology / Q000276:immunology / Q000517:prevention & control*; D014969:Xanthine Oxidase / Q000032:analysis
D015703:Antigens, CD; C497857:CD46 protein, human; D018958:CD55 Antigens; D018957:CD59 Antigens; D003169:Complement Inactivator Proteins; C497859:Mcp protein, mouse; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D009195:Peroxidase; D014969:Xanthine Oxidase