Expression and regulation of tissue inhibitor of metalloproteinase-1 and matrix metalloproteinases by intestinal myofibroblasts in inflammatory bowel disease - PubMed (original) (raw)

Expression and regulation of tissue inhibitor of metalloproteinase-1 and matrix metalloproteinases by intestinal myofibroblasts in inflammatory bowel disease

Brian C McKaig et al. Am J Pathol. 2003 Apr.

Abstract

Intestinal fibrosis and strictures frequently occur in Crohn's disease but not ulcerative colitis. We have recently shown that, compared to myofibroblasts obtained from normal and ulcerative colitis tissue, myofibroblasts isolated from fibrotic Crohn's disease mucosal samples express significantly lower amounts of transforming growth factor (TGF)-beta 3, but the expression of TGF-beta 2 was significantly greater. We now report that in myofibroblast cultures established from fibrotic Crohn's disease mucosal samples there is significantly higher constitutive expression of tissue inhibitor of metalloproteinase (TIMP)-1 compared to similar cells isolated from normal or ulcerative colitis tissue. Myofibroblasts derived from normal mucosa and from mucosa affected by ulcerative colitis or Crohn's disease also expressed matrix metalloproteinase (MMP)-1, MMP-2, and MMP-3 but did not express MMP-9. Recombinant (r) TGF-beta 1 and rTGF-beta 2, but not rTGF-beta 3, induced expression of TIMP-1 in normal intestinal myofibroblasts. These studies illustrate a potential mechanism by which differential expression of isoforms of TGF-beta may lead to excessive deposition of extracellular matrix and stricture formation via TIMP-1-mediated inhibition of MMP activity.

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Figures

Figure 1.

Figure 1.

Normal, ulcerative colitis (UC), and Crohn’s disease (CD) intestinal myofibroblasts express mRNA transcripts for MMP-1, MMP-2, and MMP-3 (lanes 2 to 4, respectively) and TIMP-1 and TIMP-2 (lanes 6 and 7, respectively). No MMP-9 mRNA was detected in any of the myofibroblasts (lane 5). Specificity of the PCR products was confirmed by DNA sequence analysis. One hundred-bp markers are shown in the left lane of the figure. The figure shown is representative of six separate experiments.

Figure 2.

Figure 2.

TIMP-1 concentrations in conditioned media of myofibroblast cultures established from normal, ulcerative colitis (UC), and Crohn’s disease (CD) mucosal samples. The samples contained equivalent total protein concentrations and TIMP-1 levels were assessed using specific ELISA.

Figure 3.

Figure 3.

Expression by Western blot analysis of TIMP-1 and TIMP-2 in conditioned media of normal, ulcerative colitis (UC), and Crohn’s disease (CD) intestinal myofibroblast cultures. Samples containing equal amounts of protein were applied to each lane and separated by SDS-polyacrylamide gel electrophoresis. After transfer to polyvinylidene difluoride membranes, immunostaining was performed using antibodies specific to TIMP-1 and TIMP-2. In contrast to TIMP-2, this representative Western blot shows greater expression of TIMP-1 in conditioned medium of Crohn’s disease intestinal myofibroblasts.

Figure 4.

Figure 4.

Representative Western blot showing expression of only the latent forms of MMP-1 (55 kd) and MMP-3 (66 kd) by normal myofibroblasts. Similar patterns were seen for myofibroblast cultures derived form ulcerative colitis and Crohn’s disease mucosa.

Figure 5.

Figure 5.

Representative gelatin zymogram showing expression of both active and latent forms of MMP-2, but no expression of MMP-9 by normal, ulcerative colitis, and Crohn’s disease myofibroblasts. Lane 1 shows a positive control, lanes 2 to 4 show normal MFCM, lanes 5 to 7 show UC MFCM, and lanes 8 to 10 show Crohn’s disease MFCM. No bands corresponding to the 92-kd MMP-9 are seen, however in all cultures, both 72-kd (latent) and 62-kd (active) bands of MMP-2 can be seen.

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References

    1. McAlindon ME, Gray T, Galvin A, Sewell HF, Podolsky DK, Mahida YR: Differential lamina propria cell migration via basement membrane pores of inflammatory bowel disease mucosa. Gastroenterology 1998, 115:841-848 - PubMed
    1. Graham MF: Pathogenesis of intestinal strictures in CD—an update. Inflamm Bowel Dis 1995, 1:220-227 - PubMed
    1. Nagase H, Woessner JF, Jr: Matrix metalloproteinases. J Biol Chem 1999, 274:21491-21494 - PubMed
    1. Gomez DE, Alonso DF, Yoshiji H, Thorgeirsson UP: Tissue inhibitors of metalloproteinases: structure, regulation and biological functions. Eur J Cell Biol 1997, 74:111-122 - PubMed
    1. Brew K, Dinakarpandian D, Nagase H: Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochim Biophys Acta 2000, 1477:267-283 - PubMed

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