Kupffer cell-derived 95kDa type IV collagenase/gelatinase : characterisation, regulation and role in liver disease
Kupffer cell-derived 95kDa type IV collagenase/gelatinase : characterisation, regulation and role in liver disease
Kupffer cells (hepatic macrophages) play an important role in host defence but have also been strongly implicated in the pathogenesis of liver injury. This thesis describes the release of 95kDa type IV collagenase/gelatinase (gelatinase B) from Kupffer cells, its characterisation, regulation and potential role in liver disease.
Kupffer cells prepared from normal rat and human liver released gelatinase B after 2 days in culture. The partially purified enzyme (from gelatin-sepharose and gel filtration chromatography of Kupffer cell conditioned medium (KCCM)) was a zinc-dependent matrix metalloproteinase with activity against native types III, IV and V collagens and gelatin. Activity was inhibited by the specific tissue inhibitor of metalloproteinases-1 (TIMP-1) and the non-specific protease scavenger α-2 macroglobulin. It was released as a proenzyme which can be activated in vitro to 75-88, 75 and 65kDa forms by organomercurials (APMA) via self cleavage. The substrate specificity of gelatinase B indicates that it has the ability to degrade the basement membrane-like matrix in the normal hepatic subendothelial space of Disse (which depends on type IV collagen for structural integrity and contains type III collagen). As with neutrophils and tumour cells release of this enzyme may facilitate cellular migration across basement membrane matrices. However, it may also play a pathogenic role in liver injury and fibrosis; degradation of the normal subendothelial matrix causes hepatocyte and endothelial cell dysfunction along with activation of fat-storing cells to a myofibroblastic phenotype.
Release of gelatinase B from cultured Kupffer cells was increased by phorbol ester (at the mRNA level), endotoxin and zymosan. TNFα, IL-1 and TGFβ had no consistent effect on release of gelatinase B, but increased release of IL-6. These studies support a role early in acute injury but not in secondary cytokine-mediated events.
University of Southampton
Winwood, Paul John
87c86a3d-5c78-4323-bbed-0f32c1c6bd68
1993
Winwood, Paul John
87c86a3d-5c78-4323-bbed-0f32c1c6bd68
Winwood, Paul John
(1993)
Kupffer cell-derived 95kDa type IV collagenase/gelatinase : characterisation, regulation and role in liver disease.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
Kupffer cells (hepatic macrophages) play an important role in host defence but have also been strongly implicated in the pathogenesis of liver injury. This thesis describes the release of 95kDa type IV collagenase/gelatinase (gelatinase B) from Kupffer cells, its characterisation, regulation and potential role in liver disease.
Kupffer cells prepared from normal rat and human liver released gelatinase B after 2 days in culture. The partially purified enzyme (from gelatin-sepharose and gel filtration chromatography of Kupffer cell conditioned medium (KCCM)) was a zinc-dependent matrix metalloproteinase with activity against native types III, IV and V collagens and gelatin. Activity was inhibited by the specific tissue inhibitor of metalloproteinases-1 (TIMP-1) and the non-specific protease scavenger α-2 macroglobulin. It was released as a proenzyme which can be activated in vitro to 75-88, 75 and 65kDa forms by organomercurials (APMA) via self cleavage. The substrate specificity of gelatinase B indicates that it has the ability to degrade the basement membrane-like matrix in the normal hepatic subendothelial space of Disse (which depends on type IV collagen for structural integrity and contains type III collagen). As with neutrophils and tumour cells release of this enzyme may facilitate cellular migration across basement membrane matrices. However, it may also play a pathogenic role in liver injury and fibrosis; degradation of the normal subendothelial matrix causes hepatocyte and endothelial cell dysfunction along with activation of fat-storing cells to a myofibroblastic phenotype.
Release of gelatinase B from cultured Kupffer cells was increased by phorbol ester (at the mRNA level), endotoxin and zymosan. TNFα, IL-1 and TGFβ had no consistent effect on release of gelatinase B, but increased release of IL-6. These studies support a role early in acute injury but not in secondary cytokine-mediated events.
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Published date: 1993
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Local EPrints ID: 462460
URI: http://eprints.soton.ac.uk/id/eprint/462460
PURE UUID: f4357eb5-77c9-4d38-8d52-018502d77716
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Date deposited: 04 Jul 2022 19:08
Last modified: 23 Jul 2022 01:07
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Author:
Paul John Winwood
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