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Gut. Jul 2000; 47(1): 63–73.
PMCID: PMC1727961

Differential expression of matrix metalloproteinases and their tissue inhibitors in colon mucosa of patients with inflammatory bowel disease

Abstract

BACKGROUND/AIMS—Alterations in synthesis and breakdown of extracellular matrix components are known to play a crucial role in tissue remodelling during inflammation and wound healing. Degradation of collagens is highly regulated by a cascade of matrix metalloproteinases (MMPs). The current study was therefore designed to determine gene expression patterns of MMPs and their tissue inhibitors (TIMPs) in single endoscopic biopsies of patients with inflammatory bowel disease (IBD).
PATIENTS/METHODS—mRNA expression was measured by quantitative competitive polymerase chain reaction (PCR) in biopsies from patients with ulcerative colitis (n=21) and Crohn's disease (n=21). Protein expression was analysed by western blotting and immunohistochemistry.
RESULTS—MMP-2, MMP-14, and TIMP-1 mRNAs were marginally increased in inflamed, but 9-12-fold increased in ulcerated colonic mucosa in IBD whereas TIMP-2 mRNA expression remained unchanged. MMP-1 and MMP-3 mRNA expression correlated well with the histological degree of acute inflammation, resulting in more than 15-fold increased MMP-1 and MMP-3 mRNA levels in inflamed versus normal colon samples from patients with ulcerative colitis and Crohn's disease.
CONCLUSION—Profound overexpression of MMP-1 and MMP-3 mRNA transcripts suggests an important role for these enzymes in the process of tissue remodelling and destruction in inflammatory bowel disease.


Keywords: matrix metalloproteinases; polymerase chain reaction; extracellular matrix; Crohn's disease; ulcerative colitis.

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Selected References

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Figures and Tables

Figure 1
Evaluation of equal amplification efficiencies of the MMP-1 PCR product and the MMP-1 competitor construct. Purified MMP-1 PCR product (+; 247 bp) and the MMP-1 competitor construct (#; 222 bp) were mixed in an OD ratio of 1:1 (lane 1) ...
Figure 2
Expression of MMP-1, -2, -3, -14, the tissue inhibitors TIMP-1 and TIMP-2, collagen type III, and GAPDH mRNA in colon biopsies from non-inflamed areas and inflamed areas of patients with Crohn's disease (CD) and ulcerative colitis ...
Figure 3
Increased protein expression of MMP-1, MMP-2, and MMP-3 in inflamed (i) versus non-inflamed (n) colon biopsies from patients with IBD. Endoscopic biopsies from macro- and microscopically normal (lanes 1, 3, 5) and inflamed (lanes 2, 4, 6) ...
Figure 4
Molar ratios of matrix metalloproteinase to TIMP-1 specific cDNA derived from reverse transcribed mRNA isolated from colon biopsies of non-inflamed areas and inflamed areas of patients with Crohn's disease (CD) and ulcerative colitis (UC). Competitive ...
Figure 5
Relationship between histological degree of acute inflammation in colon biopsies from IBD patients and mRNA expression of MMPs, TIMP-1, and collagen type III measured by competitive PCR. Scattergrams show the ratio of gene specific transcripts to β-actin ...
Figure 6
Immunohistochemical staining of MMP-1 in inflamed and non-inflamed colon of a patient with ulcerative colitis. Section of paraffin embedded biopsies from non-inflamed (A, C) and inflamed (B, D-F) colon were stained with haematoxylin/eosin (A, B) and ...

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