In vitro evidence for a bacterial pathogenesis of equine laminitis.
Utilizing an in vitro laminitis explant model, we have investigated how bacterial broth cultures and purified bacterial proteases activate matrix metalloproteinases (MMPs) and alter structural integrity of cultured equine lamellar hoof explants. Four Gram-positive Streptococcus spp. and three Gram-negative bacteria all induced a dose-dependent activation of MMP-2 and MMP-9 and caused lamellar explants to separate. MMP activation was deemed to have occurred if a specific MMP inhibitor, batimastat, blocked MMP activity and prevented lamellar separation. Thermolysin and streptococcal pyrogenic exotoxin B (SpeB) both separated explants dose-dependently but only thermolysin was inhibitable by batimastat or induced MMP activation equivalent to that seen with bacterial broths. Additionally, thermolysin and broth MMP activation appeared to be cell dependent as MMP activation did not occur in isolation. These results suggest the rapid increase in streptococcal species in the caecum and colon observed in parallel with carbohydrate induced equine laminitis may directly cause laminitis via production of exotoxin(s) capable of activating resident MMPs within the lamellar structure. Once activated, these MMPs can degrade key components of the basement membrane (BM) hemidesmosome complex, ultimately separating the BM from the epidermal basal cells resulting in the characteristic laminitis histopathology of hoof lamellae. While many different causative agents have been evaluated in the past, the results of this study provide a unifying aetiological mechanism for the development of carbohydrate induced equine laminitis.
Department of Companion Animal Sciences, School of Veterinary Science, The University of Queensland, St. Lucia, Qld 4072, Australia.
B A
BA
Mungall
M
M
Kyaw-Tanner
C C
CC
Pollitt
D000818:Animals; D001426:Bacterial Proteins; D003546:Cysteine Endopeptidases / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004789:Enzyme Activation; D005534:Foot Diseases / Q000201:enzymology / Q000382:microbiology / Q000473:pathology / Q000662:veterinary*; D006090:Gram-Negative Bacteria / Q000472:pathogenicity; D006651:Histocytochemistry / Q000662:veterinary; D006724:Hoof and Claw / Q000201:enzymology / Q000382:microbiology* / Q000473:pathology; D006734:Horse Diseases / Q000201:enzymology / Q000382:microbiology* / Q000473:pathology; D006736:Horses; D007794:Lameness, Animal / Q000201:enzymology / Q000382:microbiology* / Q000473:pathology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D016985:Streptococcus bovis / Q000472:pathogenicity*; D013820:Thermolysin / Q000494:pharmacology; D013876:Thiophenes / Q000494:pharmacology
D001426:Bacterial Proteins; D011480:Protease Inhibitors; D013876:Thiophenes; D010649:Phenylalanine; C080985:batimastat; D003546:Cysteine Endopeptidases; C042477:streptopain; D020778:Matrix Metalloproteinase 2; D013820:Thermolysin; D020780:Matrix Metalloproteinase 9