Elastin accumulation is regulated at the level of degradation by macrophage metalloelastase (MMP-12) during experimental liver fibrosis.
Elastin has been linked to maturity of liver fibrosis. To date, the regulation of elastin secretion and its degradation in liver fibrosis has not been characterized. The aim of this work was to define elastin accumulation and the role of the paradigm elastase macrophage metalloelastase (MMP-12) in its turnover during fibrosis. Liver fibrosis was induced by either intraperitoneal injections of carbon tetrachloride (CCl(4) ) for up to 12 weeks (rat and mouse) or oral administration of thioacetamide (TAA) for 1 year (mouse). Elastin synthesis, deposition, and degradation were investigated by immunohistochemistry, quantitative polymerase chain reaction (qPCR), western blotting, and casein zymography. The regulation of MMP-12 elastin degradation was defined mechanistically using CD11b-DTR and MMP-12 knockout mice. In a CCl(4) model of fibrosis in rat, elastin deposition was significantly increased only in advanced fibrosis. Tropoelastin expression increased with duration of injury. MMP-12 protein levels were only modestly changed and in coimmunoprecipitation experiments MMP-12 was bound in greater quantities to its inhibitor TIMP-1 in advanced versus early fibrosis. Immunohistochemistry and macrophage depletion experiments indicated that macrophages were the sole source of MMP-12. Exposure of CCl(4) in MMP-12(-/-) mice led to a similar degree of overall fibrosis compared to wildtype (WT) but increased perisinusoidal elastin. Conversely, oral administration of TAA caused both higher elastin accumulation and higher fibrosis in MMP-12(-/-) mice compared with WT.


CONCLUSIONS
Elastin is regulated at the level of degradation during liver fibrosis. Macrophage-derived MMP-12 regulates elastin degradation even in progressive experimental liver fibrosis. These observations have important implications for the design of antifibrotic therapies.
MRC/UoE Centre for Inflammation Research, University of Edinburgh, Edinburgh, UK. apellico@staffmail.ed.ac.uk
Antonella
A
Pellicoro
Rebecca L
RL
Aucott
Prakash
P
Ramachandran
Andrew J
AJ
Robson
Jonathan A
JA
Fallowfield
Victoria K
VK
Snowdon
Stephen N
SN
Hartland
Madeleine
M
Vernon
Jeremy S
JS
Duffield
R Christopher
RC
Benyon
Stuart J
SJ
Forbes
John P
JP
Iredale
D000818:Animals; D002251:Carbon Tetrachloride / Q000633:toxicity; D004549:Elastin / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008106:Liver Cirrhosis, Experimental / Q000378:metabolism*; D008264:Macrophages / Q000201:enzymology*; D053508:Matrix Metalloproteinase 12 / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D051381:Rats; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D014333:Tropoelastin / Q000096:biosynthesis
D019715:Tissue Inhibitor of Metalloproteinase-1; D014333:Tropoelastin; D004549:Elastin; D002251:Carbon Tetrachloride; D053508:Matrix Metalloproteinase 12