Function of tissue inhibitor of metalloproteinase-1 in liver fibrosis
Function of tissue inhibitor of metalloproteinase-1 in liver fibrosis
This work aimed to further the current knowledge of the role of TIMP-1 in the development of liver fibrosis. The first aim was to develop and validate a method of quantitating TIMP-1 mRNA expression of TIMP-1 in human liver biopsies using RT-PCR. Utilising this method, the expression in human liver biopsies was shown to increase compared to normal, in each of the fibrotic disorders tested, namely primary biliary cirrhosis, primary sclerosing cholangitis and biliary atresia, supporting the concept that TIMP-1 could be an important factor in fibrosis development.
At the time of commencing this thesis, antibodies to rat MMP-13 and rat TIMP-1 were not available to use for the immunolocalisation of these proteins in the rat models of fibrosis. The aim of this thesis was to develop anti-rat TIMP-1 and MMP-13 antibodies to complement the Q-RT-PCR work. This thesis describes the synthesis of anti-rat MMP-13 and TIMP-1 antigens for immunisation of rabbits, the purification and specificity testing of the resulting sera. Dot and Western blot analysis, as well as immunostaining of isolated rat hepatic stellate cells, have revealed the presence of antibodies which may prove useful for immunodetection studies.
Access to TIMP-1 deleted mice provided an ideal opportunity to examine the functional activities of TIMP-1 deleted hepatic stellate cells following activation in vitro. Histological analysis of liver sections following the administration of the fibrogenic agent CCL4, revealed that removal of TIMP-1 from the system did not affect progression of fibrosis.
Measurement of the rate of collagen deposition, proliferation and gelatinase activity of hepatic stellate cells during activation by culture on plastic, showed no significant differences between the TIMP-1 deleted and wild type genotypes.
These results suggest that although the upregulation of TIMP-1 has been demonstrated during the progression of liver fibrosis, its presence may not be an essential requirement of this disease process, as shown by the murine TIMP-1 knockout model.
University of Southampton
Pickering, Judith Ann
053a7c95-8a8c-4847-8bb7-6b2cceb0322b
1998
Pickering, Judith Ann
053a7c95-8a8c-4847-8bb7-6b2cceb0322b
Pickering, Judith Ann
(1998)
Function of tissue inhibitor of metalloproteinase-1 in liver fibrosis.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
This work aimed to further the current knowledge of the role of TIMP-1 in the development of liver fibrosis. The first aim was to develop and validate a method of quantitating TIMP-1 mRNA expression of TIMP-1 in human liver biopsies using RT-PCR. Utilising this method, the expression in human liver biopsies was shown to increase compared to normal, in each of the fibrotic disorders tested, namely primary biliary cirrhosis, primary sclerosing cholangitis and biliary atresia, supporting the concept that TIMP-1 could be an important factor in fibrosis development.
At the time of commencing this thesis, antibodies to rat MMP-13 and rat TIMP-1 were not available to use for the immunolocalisation of these proteins in the rat models of fibrosis. The aim of this thesis was to develop anti-rat TIMP-1 and MMP-13 antibodies to complement the Q-RT-PCR work. This thesis describes the synthesis of anti-rat MMP-13 and TIMP-1 antigens for immunisation of rabbits, the purification and specificity testing of the resulting sera. Dot and Western blot analysis, as well as immunostaining of isolated rat hepatic stellate cells, have revealed the presence of antibodies which may prove useful for immunodetection studies.
Access to TIMP-1 deleted mice provided an ideal opportunity to examine the functional activities of TIMP-1 deleted hepatic stellate cells following activation in vitro. Histological analysis of liver sections following the administration of the fibrogenic agent CCL4, revealed that removal of TIMP-1 from the system did not affect progression of fibrosis.
Measurement of the rate of collagen deposition, proliferation and gelatinase activity of hepatic stellate cells during activation by culture on plastic, showed no significant differences between the TIMP-1 deleted and wild type genotypes.
These results suggest that although the upregulation of TIMP-1 has been demonstrated during the progression of liver fibrosis, its presence may not be an essential requirement of this disease process, as shown by the murine TIMP-1 knockout model.
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Published date: 1998
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Local EPrints ID: 463277
URI: http://eprints.soton.ac.uk/id/eprint/463277
PURE UUID: 60ce7a67-e5de-4927-988f-766a58606b93
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Date deposited: 04 Jul 2022 20:48
Last modified: 23 Jul 2022 01:09
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Author:
Judith Ann Pickering
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