Lung Collagenase Inhibitors and Spontaneous and Latent Collagenase Activity in Idiopathic Pulmonary Fibrosis and Hypersensitivity Pneumonitis

doi:10.1378/chest.96.5.1115

Chest

Volume 96, Issue 5, November 1989, Pages 1115-1119

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In order to analyze the mechanisms involved in the decreased collagenolytic activity previously observed in interstitial lung fibrosis, we studied the inhibitory collagenase activity and the latent activable collagenase in lung samples from five patients with IPF, six with HP, and three control subjects. Our results showed that in both diseases, the inhibitor levels were significantly higher than in control subjects. Findings suggest that in IPF low amounts of collagenase plus excessive enzyme-inhibitors may be operating to decrease collagen catabolism. In contrast, HP lungs seem to contain adequate amounts of the enzyme but higher levels of inhibitors play a role in the abnormal degradation observed in some patients.

Section snippets

Study Population

We studied five patients with IPF, four women and one man, aged 31 to 57 years old, mean age: 42.4 years; and six female patients with chronic HP with age ranging from 23 to 45 years old, with a mean of 31.3 years. All patients of both groups fulfilled the clinical criteria for diagnosis described elsewhere.5, 13, 14, 15 The diagnosis was confirmed by the morphologic findings in the tissue samples obtained through an open lung biopsy. Briefly, lung tissue from IPF patients showed diffuse

Results

The results of endogeneous collagenolytic activity revealed without activation are shown in Figure 1. The IPF and HP tissue samples presented a similar behavior as previously described.5, 6 Thus, decreased values of collagen degradation were found in three out of five IPF patients with an average of 0.15±0.08 µg of collagen degraded per milligram of collagen incubated per hour. In control subjects, the average value was 0.24±0.04. On the other hand, an heterogeneous behavior with high, normal

Discussion

Collagenolytic activity plays a crucial role in the modulation of collagen tissue turnover, and several previous studies performed in our laboratory have demonstrated that a decrease in collagenolysis usually occurs during the development of experimental and human interstitial lung fibrosis.5, 6, 7, 8

In order to go deep into the possible mechanisms involved in this alteration, we analyzed both the presence of latent collagenase and inhibitory activity in lung tissue from IPF and HP

Latent

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Subpopulations of T cells in lung biopsies from patients with pigeon breeder's disease.

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Cited by (38)

Extracellular matrix in lung development, homeostasis and disease
2018, Matrix Biology
Citation Excerpt :
However, the mechanisms regulating matrix degradation, and how these mechanisms are perturbed in chronic lung disease, have received far less attention [160]. An impaired tissue degradative environment has been observed in both pulmonary fibrosis and fibrosis in other organ systems [172,173]. In IPF patients, for example, there is an imbalance between the production of MMPs and TIMPs with an increase in the ratio of TIMPs:MMPs at the site of scar formation [173].
The lung's unique extracellular matrix (ECM), while providing structural support for cells, is critical in the regulation of developmental organogenesis, homeostasis and injury-repair responses. The ECM, via biochemical or biomechanical cues, regulates diverse cell functions, fate and phenotype. The composition and function of lung ECM become markedly deranged in pathological tissue remodeling. ECM-based therapeutics and bioengineering approaches represent promising novel strategies for regeneration/repair of the lung and treatment of chronic lung diseases. In this review, we assess the current state of lung ECM biology, including fundamental advances in ECM composition, dynamics, topography, and biomechanics; the role of the ECM in normal and aberrant lung development, adult lung diseases and autoimmunity; and ECM in the regulation of the stem cell niche. We identify opportunities to advance the field of lung ECM biology and provide a set recommendations for research priorities to advance knowledge that would inform novel approaches to the pathogenesis, diagnosis, and treatment of chronic lung diseases.
Decreased capacity of asthmatic bronchial fibroblasts to degrade collagen
2001, Matrix Biology
The mechanisms of fibrillar collagen accumulation in asthmatic bronchi remain unclear, an imbalance between synthesis and degradation of collagen may be implicated in this process. The aim of this study was to compare the capacities of normal (BNF) and asthmatic (BAF) bronchial fibroblasts to degrade collagen. Metalloproteinases and their inhibitors were measured by ELISA, types I and III procollagen synthesis was determined by liquid RIA and, finally, zymography was used to assess the presence of active and latent forms of MMPs. The capacity of fibroblasts to degrade collagen coated onto latex beads was evaluated by flow cytometry. Our results showed that MMP-2 secretion was significantly higher in BNF when compared to BAF and this was confirmed by gelatin zymography. In BNF culture, TIMP-1 and MMP-1 secretions positively correlated with types I and III procollagen synthesis. However, in BAF, this correlation was negative. This suggests that a balance exists between collagen synthesis and degradation in BNF and that this balance is compromised in BAF. On the other hand, BAF did show significantly reduced capacity to degrade collagen when compared to that of BNF. This reduced phagocytic activity was not associated with a decrease in collagen receptor expression. This study establishes for the first time that a relationship exists between metalloproteinases enzyme dysregulation and the reduced capacity of asthmatic bronchial fibroblast to degrade collagen. These events may shed light on why accumulation of collagen can be observed in asthmatic airways.
Induction of MMP-9 mediated gelatinolytic activity in human monocytic cells by cell wall components of Mycobacterium tuberculosis
2000, Microbial Pathogenesis
Mycobacterium tuberculosis (Mtb) infection induces the expression of host matrix metalloÍproteinases (MMPs) capable of tissue degradation. We show that infection of mice with Mtb results in differential expression of MMPs in the lung. MMP-9 activity increased by week 1 post-infection, while MMP-2 activity increased after week 2. RT-PCR analysis for gene expression of gelatinases and their respective inhibitors showed: a small increase in MMP-9 by week 1, no change in TIMP-1 and MMP-2, and a significant decrease in TIMP-2 by week 4. The increase in MMP-2 could be due to a decrease in TIMP-2 expression. Addition of 4-aminophenylmercuric acid to lung extracts increased MMP-9 activity, suggesting that its regulation could be due to endogenous activation by proteases. In vitro, attenuated and virulent Mtb strains equally induced MMP-9 expression in U937 monocytes. The inducer of MMP-9 in Mtb was present in culture filtrates, and was active after paraformaldehyde fixation. LAM stimulated MMP-9 expression in THP-1 cells, but not U937 cells. However, LAM-free extracts also induced MMP-9 activity in THP-1 cells. Fractionation of Mtb extracts by chromatography revealed fractions of 17 and 156 kDa with MMP-9 inducing activity. In conclusion, LAM and other components of Mtb induce the expression of MMP-9.
Tissue inhibitors of metalloproteinases in liver fibrosis
1997, International Journal of Biochemistry and Cell Biology
Liver fibrosis and its end stage sequelae cirrhosis represent a major worldwide health problem. By definition progressive fibrosis occurs when the rate of matrix synthesis exceeds matrix degradation. Considerable evidence suggests that the hepatic stellate cell is central to the fibrotic process. During liver injury these cells transform from a quiescent retinoid filled phenotype to a proliferative myofibroblast like cell. In this ‘activated’ phenotype the HSC is the major source of the interstitial collagens, which characterize fibrosis. Recent work suggests that the HSCs are also a source of matrix degrading metalloproteinase (MMPs), indicating that, together with other cells, hepatic stellate cells (HSC) could participate in matrix remodelling. However, HSC activation in tissue culture models and in vivo is also associated with expression of the powerful MMP inhibitors: tissue inhibitors of metalloproteinases 1 and 2 (TIMP-1 and TIMP-2). TIMP expression has also been demonstrated in fibrotic human liver disease and animal models of liver fibrosis. TIMPs 1 and 2 may therefore promote progression of hepatic fibrosis through inhibition of matrix degradation.
Tissue inhibitor of metalloproteinase-1 messenger RNA expression is enhanced relative to interstitial collagenase messenger RNA in experimental liver injury and fibrosis
1996, Hepatology
Liver fibrosis results from a relative imbalance between synthesis and degradation of matrix proteins. We have previously described release of the protein collagenase inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), by culture-activated human hepatic stellate cells (HSCs). In this study, we have investigated the relative expression of TIMP-1 and interstitial collagenase in culture-activated rat HSCs and rat models of liver injury and fibrosis. The complementary DNA (cDNA) for rat TIMP-1 was obtained by homology polymerase chain reaction (PCR) and sequenced. By Northern analysis using this probe, TIMP-1 messenger RNA (mRNA) expression was up-regulated with HSC activation by culture on plastic as defined by cellular expression of procollagen-1. Interstitial collagenase mRNA was expressed in early 1. Interstitial collagenase mRNA was expressed in early culture (<4 days) but became undetectable in more activated cells (7-21 days). By activity assay of serum-free cell-conditioned media, TIMP-1 was found to be released in increasingly concentrations with duration of culture on plastic. Expression of TIMP-1 interstitial collagenase, and procollagen-1 mRNAs were studied in rat models of biliary and parenchymal injury (bile duct ligation and CC1₄ administration) by ribonuclease protein assay. TIMP-1 mRNA expression was increased at 6, 24 hours, and 3 days after bile duct ligation and was also shown to rise in acute CC1₄ liver injury and remain elevated as the liver became fibrotic. TIMP-1 expression preceded procollagen-1 expression in both models. In contrasts, interstitial collagenase mRNA levels remained similar to control values throughout both models of liver injury. Total cellular RNA from hepatocytes, HSCs, and kupffer cells freshly isolated from livers after acute CC1₄ injury was subjected to Northern analysis. TIMP-1 transcripts were observed in nonparenchymal cells only. We suggest that increased expression of TIMP-1 relative to interstitial collagenase by HSCs may promote progression of liver fibrosis in these rat models by preventing degradation of secreted collagens. (Hepatology 1996 Jul;24(1):176-84)
Development of an enzyme-linked immunosorbent assay to measure total TIMP-1 (Free TIMP-1 and TIMP-1 in combination with matrix-metalloproteinases) and measurement of TIMP 1 and CRP in serum
1995, Clinica Chimica Acta
A panel of six monoclonal antibodies (MAbs) was raised against purified human fibroblast tissue inhibitor of metalloproteinase-1 (TIMP-1) and characterised. All possible antibody pairs were tested for their suitability as capture and revealing antibodies in a two-site enzymelinked immunosorbent assay (ELISA) to measure total TIMP-1 (both free TIMP-1 and TIMP-1 together with matrix metalloproteinases (MMPs)). Using the best combination of MAbs the assay was optimised. The sensitivity of detection of the assay was 1.4 ng/ml, and inter- and intra-assay coefficients of variation were between 10.4–13.7% and 8.8–9.7%, respectively. Dilution series of human cerebrospinal and synovial fluids, plasma and sera paralleled those of the TIMP-1 standard curve indicating that the immunoreactivity detected in these samples was authentic TIMP-1. TIMP-2 shows no detectable cross reactivity in this assay confirming that this ELISA is specific for TIMP-1. The levels of total TIMP-1 and collagenase were measured in conditioned medium from A2058 human melanoma cells cultured in the absence or presence of human recombinant interleukin-1 α (hrlL-lα). Total TIMP-1 was also measured in serum samples with known C-reactive protein (CRP) (n = 100) and α₁ antichymotrypsin (ACT) (n = 52) concentrations; no correlation was found between TIMP-1 levels and either of these acute phase reactants although the levels of TIMP-1 were raised when compared to normal sera. This ELISA provides a rapid and convenient procedure for the quantitation of total TIMP-1 in human biological fluids and supernatants from cultured cell lines.

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Chest

Clinical InvestigationsLung Collagenase Inhibitors and Spontaneous and Latent Collagenase Activity in Idiopathic Pulmonary Fibrosis and Hypersensitivity Pneumonitis

Section snippets

Study Population

Results

Discussion

An introduction to the interstitial lung diseases. In: Fulmer JD. ed. Clinics in chest medicine: symposium on interstitial lung diseases

Interstitial lung disease: current concepts of pathogenesis, staging and therapy.

Am J Med

Lung collagen: more than scaffolding.

Thorax

Concentration, biosynthesis and degradation of collagen in idiopathic pulmonary fibrosis.

Thorax

Lung collagen metabolism and the clinical course of hypersensitivity pneumonitis.

Chest

Experimental pulmonary fibrosis induced by paraquat plus oxygen in rats: a morphologic and biochemical sequential study.

Exp Molec Pathol

Regulation of the mammalian collagenases.

Collagen Rel Res

What controls collagen resorption in vivo?

Med Hypotheses

Human alveolar macrophages produce a fibroblast-like collagenase and collagenase inhibitor.

J Clin Invest

Human fibroblast collagenase: glycosylation and tissue-specific levels of enzyme synthesis.

Proc Natl Acad Sci USA

Relationship between lung inflammation or fibrosis and frequency dependence of compliance in interstitial pulmonary diseases.

Respiration

Subpopulations of T cells in lung biopsies from patients with pigeon breeder's disease.

Lung

Clinical Investigations
Lung Collagenase Inhibitors and Spontaneous and Latent Collagenase Activity in Idiopathic Pulmonary Fibrosis and Hypersensitivity Pneumonitis