Matrix metalloproteinases in necrotising enterocolitis.
Elevated cytokines, especially TNF-alpha, have been implicated in the pathogenesis of necrotising enterocolitis (NEC). We have previously shown that TNF-alpha drives the production of matrix degrading enzymes, the matrix metalloproteinases (MMPs), in the gut wall. In this study we have therefore investigated the role of MMPs in the pathogenesis of NEC in neonates. Nine newborn infant nonnecrotic resected bowels with confirmed NEC were studied and 8 newborn infants with neonatal bowel obstructions were used as controls. Immunostaining was used to identify the numbers of monocytes, macrophages, neutrophils, and T cells in the tissue. We used quantitative, competitive RT-PCR to analyze the number of TNF-alpha, IFN-gamma, MMP, and TIMP mRNA transcripts and western blotting to analyze MMP and TIMP protein production. Double labeling (immunostaining and in situ hybridization) was used to identify the phenotype of MMP mRNA expressing cells. We found increased numbers of monocytes, macrophages, and neutrophils in NEC tissue compared with controls. The number of T cells was unexpectedly low in NEC as was the number of IFN-gamma transcripts in comparison with the control samples. Increased numbers of transcripts for TNF-alpha were detected in NEC tissue, as was mRNA expression and protein production for stromelysin-1 and TIMP-1 but not collagenase, gelatinases, or TIMP-2. The cellular source of stromelysin-1 in NEC was alpha-smooth muscle actin positive cells. These results suggest that stromelysin-1, which has the ability to degrade the mucosal extra-cellular matrix, may be responsible for the extensive tissue injury in infants with NEC.
Division of Infection, Inflammation and Repair, Mailpoint 813, Level E, South Academic Block, Southampton General Hospital, Tremona Road, Southampton, SO16 6YD, UK. s.pender@soton.ac.uk
Sylvia Lin Foon
SL
Pender
Christian
C
Braegger
Ute
U
Gunther
Giovanni
G
Monteleone
Martin
M
Meuli
Detlef
D
Schuppan
Thomas Thornton
TT
Macdonald
D020345:Enterocolitis, Necrotizing / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007231:Infant, Newborn; D007371:Interferon-gamma / Q000235:genetics; D007413:Intestinal Mucosa / Q000201:enzymology* / Q000276:immunology; D020778:Matrix Metalloproteinase 2 / Q000235:genetics; D019278:Matrix Metalloproteinase 3 / Q000235:genetics*; D032389:Myocytes, Smooth Muscle / Q000201:enzymology / Q000276:immunology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D019715:Tissue Inhibitor of Metalloproteinase-1; D014409:Tumor Necrosis Factor-alpha; D019716:Tissue Inhibitor of Metalloproteinase-2; D007371:Interferon-gamma; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2