Increased production of matrix metalloproteinase-3 and tissue inhibitor of metalloproteinase-1 by inflamed mucosa in inflammatory bowel disease

Abstract

Inflammatory bowel diseases (IBD) are characterized by a sustained inflammatory cascade that gives rise to the release of mediators capable of degrading and modifying bowel wall structure. Our aims were (i) to measure the production of matrix metalloproteinase-3 (MMP-3), and its tissue inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), by inflamed and uninflamed colonic mucosa in IBD, and (ii) to correlate their production with that of proinflammatory cytokines and the anti-inflammatory cytokine, IL-10. Thirty-eight patients with IBD, including 25 with Crohn's disease and 13 with ulcerative colitis, were included. Ten controls were also studied. Biopsies were taken from inflamed and uninflamed regions and inflammation was graded both macroscopically and histologically. Organ cultures were performed for 18 h. Tumour necrosis factor-alpha (TNF-alpha), IL-6, IL-1beta, IL-10, MMP-3 and TIMP-1 concentrations were measured using specific immunoassays. The production of both MMP-3 and the TIMP-1 were either undetectable or below the sensitivity of our immunoassay in the vast majority of uninflamed samples either from controls or from those with Crohn's disease or ulcerative colitis. In inflamed mucosa, the production of these mediators increased significantly both in Crohn's disease (P < 0.01 and 0.001, respectively) and ulcerative colitis (P < 0.001 and 0.001, respectively). Mediator production in both cases was significantly correlated with the production of proinflammatory cytokines and IL-10, as well as with the degree of macroscopic and microscopic inflammation. Inflamed mucosa of both Crohn's disease and ulcerative colitis show increased production of both MMP-3 and its tissue inhibitor, which correlates very well with production of IL-1beta, IL-6, TNF-alpha and IL-10.

Fig. 1

Production of matrix metalloproteinase-3 (MMP-3) by colonic mucosa in organ culture. Cultures were performed for 18 h without stimulus. MMP-3 concentrations were measured in the culture medium using a specific immunoassay. The detection limit of the test was 100 pg/ml. Each point corresponds to an individual patient and represents the mean of three biopsy cultures. There was an increased production of MMP-3 in inflamed Crohn's disease (CD) and ulcerative colitis (UC) (P < 0·01 and 0·001, respectively).

Fig. 2

Production of tissue inhibitor of metalloproteinase-1 (TIMP-1) by colonic mucosa in organ culture. Cultures were performed for 18 h without stimulus. TIMP-1 concentrations were measured in the culture medium using a specific immunoassay. The detection limit of the test was 1250 pg/ml. Each point corresponds to an individual patient and represents the mean of three biopsy cultures. There was an increased production of TIMP-1 in inflamed Crohn's disease (CD) and ulcerative colitis (UC) (P < 0·001 and 0·001, respectively).

Fig. 3

Correlation between production of matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) by colonic mucosa in organ culture in Crohn's disease. Cultures were performed for 18 h without stimulus. MMP-3 and TIMP-1 concentrations were measured in the culture medium using a specific immunoassay. Each point corresponds to an individual patient and represents the mean of three biopsy cultures. Using linear regression, the two mediators were significantly correlated (r = 0·59; P < 0·01).

Fig. 4

Correlation between production of matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) by colonic mucosa in organ culture in ulcerative colitis. Cultures were performed for 18 h without stimulus. MMP-3 and TIMP-1 concentrations were measured in the culture medium using a specific immunoassay. Each point corresponds to an individual patient and represents the mean of three biopsy cultures. Using linear regression, the two mediators were significantly correlated (r = 0·93; P < 0·0001).