Phosphodiesterase inhibition mediates matrix metalloproteinase activity and the level of collagen degradation fragments in a liver fibrosis ex vivo rat model

被引：16
作者：
Sanne Skovgård Veidal
Mette Juul Nielsen
Diana Julie Leeming
Morten Asser Karsdal
机构：
[1] Nordic Bioscience A/S, 2730, Herlev
来源：
BMC Research Notes | / 5卷 / 1期
关键词：
cAMP; Ex vivo; Fibrosis; Precision-cut liver slices;
D O I：
10.1186/1756-0500-5-686
中图分类号：
学科分类号：
摘要：
Abstract. Background: Accumulation of extracellular matrix (ECM) and increased matrix metalloproteinase (MMP) activity are hallmarks of liver fibrosis. The aim of the present study was to develop a model of liver fibrosis combining ex vivo tissue culture of livers from CCl§ssub§4§ esub§ treated animals with an ELISA detecting a fragment of type III collagen generated in vitro by MMP-9 (C3M), known to be associated with liver fibrosis and to investigate cAMP modulation of MMP activity and liver tissue turnover in this model. Findings. In vivo: Rats were treated for 8 weeks with CCl§ssub§4§esub§/Intralipid. Liver slices were cultured for 48 hours. Levels of C3M were determined in the supernatants of slices cultured without treatment, treated with GM6001 (positive control) or treated with IBMX (phosphodiesterase inhibitor). Enzymatic activity of MMP-2 and MMP-9 were studied by gelatin zymography. Ex vivo: The levels of serum C3M increased 77% in the CCl§ssub§4§esub§-treated rats at week 8 (p < 0.01); Levels of C3M increased significantly by 100% in fibrotic liver slices compared to controls after 48 hrs (p < 0.01). By adding GM6001 or IBMX to the media, C3M was restored to control levels. Gelatin zymography demonstrated CCl§ssub§4§esub§-treated animals had highly increased MMP-9, but not MMP-2 activity, compared to slices derived from control animals. Conclusions: We have combined an ex vivo model of liver fibrosis with measurement of a biochemical marker of collagen degradation in the condition medium. This technology may be used to evaluate the molecular process leading to structural fibrotic changes, as collagen species are the predominant structural part of fibrosis. These data suggest that modulation of cAMP may play a role in regulation of collagen degradation associated with liver fibrosis. © 2012 Veidal et al.; licensee BioMed Central Ltd.
引用
收藏
相关论文
共 52 条
  • [1] Friedman S.L., Liver fibrosis - From bench to bedside, J Hepatol, 38, SUPPL. 1, (2003)
  • [2] Gressner O.A., Weiskirchen R., Gressner A.M., Biomarkers of liver fibrosis: Clinical translation of molecular pathogenesis or based on liver-dependent malfunction tests, Clin Chim Acta, 381, pp. 107-113, (2007)
  • [3] Bedossa P., Dargere D., Paradis V., Sampling variability of liver fibrosis in chronic hepatitis C, Hepatology, 38, pp. 1449-1457, (2003)
  • [4] Maharaj B., Maharaj R.J., Leary W.P., Cooppan R.M., Naran A.D., Pirie D., Sampling variability and its influence on the diagnostic yield of percutaneous needle biopsy of the liver, Lancet, 1, pp. 523-525, (1986)
  • [5] Veidal S.S., Bay-Jensen A.C., Tougas G., Karsdal M.A., Vainer B., Serum markers of liver fibrosis: Combining the BIPED classification and the neo-epitope approach in the development of new biomarkers, Dis Markers, 28, pp. 15-28, (2010)
  • [6] Kumar V., Tissue Renewal and Repair: Regeneration, Healing, and Fibrosis. Pathologic Basis of Disease, pp. 87-118, (2005)
  • [7] Schuppan D., Structure of the extracellular matrix in normal and fibrotic liver: Collagens and glycoproteins, Semin Liver Dis, 10, pp. 1-10, (1990)
  • [8] Schuppan D., Ruehl M., Somasundaram R., Hahn E.G., Matrix as a modulator of hepatic fibrogenesis, Semin Liver Dis, 21, pp. 351-372, (2001)
  • [9] Lochter A., Bissell M.J., An odyssey from breast to bone: Multi-step control of mammary metastases and osteolysis by matrix metalloproteinases, APMIS, 107, pp. 128-136, (1999)
  • [10] Karsdal M.A., Madsen S.H., Christiansen C., Henriksen K., Fosang A.J., Sondergaard B.C., Cartilage degradation is fully reversible in the presence of aggrecanase but not matrix metalloproteinase activity, Arthritis Res Ther, 10, (2008)
123456