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Updated: May 16, 2026
Isolation of Rat Portal Fibroblasts by In situ Liver Perfusion
Published on: June 29, 2012
Collagenolytic potential of rat liver myofibroblasts
A Jiroutová1, E Peterová, L Bittnerová
1Faculty of Medicine in Hradec Králové, Hradec Králové, Czech Republic. kanta@lfhk.cuni.cz
Abstract:
Rat liver myofibroblasts (MFB) were isolated by repeated passaging of nonparenchymal liver cell fraction. They were cultured on polystyrene Petri dishes, on fibrin or on type I collagen gels for 5 days. Quantitative RT-PCR, Western blotting, zymography and immunocytochemistry were used to study differences in cell morphology and protein expression. MFB were large and spread on plastic substrate, with prominent alpha-smooth muscle (alpha-SMA) fibres. They turned much smaller and elongated on collagen which was accompanied by the rearrangement of the cytoskeleton and a decrease in alpha-SMA and beta-actin content. Collagen gel induced the expression of a group of metalloproteinases (MMP-2, -3, -9, -13), on mRNA and protein level which resulted in the degradation of the gel. This response was accompanied by changes in the mRNA expression of cytokines of TGF-beta family, CTGF and interleukin-6, as well as of osteopontin and thrombospondin-2 that are involved in metalloproteinases (MMPs) regulation. The expression of MMPs substrates, collagen types I, IV and XII did not change or decreased. The effects of fibrin gels on MFB were milder than those of collagen. MFB assumed to deposit collagen and other ECM components in fibrotic liver, besides hepatic stellate cells, also possess a great collagenolytic potential.
Insights
Rat liver myofibroblasts cultured on collagen gels showed increased matrix-degrading enzymes, suggesting a role in liver fibrosis beyond collagen deposition. These findings highlight their collagenolytic potential.
Area of Science:
- Hepatology
- Cell Biology
- Biochemistry
Background:
- Liver fibrosis involves extracellular matrix (ECM) deposition by hepatic stellate cells and potentially other cell types.
- The role of liver myofibroblasts (MFB) in ECM remodeling during liver fibrogenesis is not fully understood.
Purpose of the Study:
- To investigate the behavior and protein expression of rat liver myofibroblasts (MFB) when cultured on different substrates.
- To determine the impact of collagen and fibrin gels on MFB morphology, cytoskeleton, and matrix-degrading enzyme production.
Main Methods:
- Isolation and culture of rat liver myofibroblasts (MFB) on polystyrene, fibrin, and type I collagen gels.
- Analysis of cell morphology and protein expression using quantitative RT-PCR, Western blotting, zymography, and immunocytochemistry.
Main Results:
- MFB exhibited distinct morphologies and cytoskeletal rearrangements on different substrates, with reduced alpha-smooth muscle (alpha-SMA) and beta-actin on collagen.
- Collagen gel culture induced significant expression of matrix metalloproteinases (MMPs: MMP-2, -3, -9, -13) at mRNA and protein levels, leading to gel degradation.
- Upregulation of cytokines (TGF-beta family, CTGF, IL-6) and MMP regulators (osteopontin, thrombospondin-2) was observed in response to collagen gels.
Conclusions:
- Rat liver myofibroblasts (MFB) possess substantial collagenolytic potential, indicated by induced MMP expression and activity on collagen gels.
- Beyond ECM deposition, MFB actively remodel the matrix, contributing to liver fibrotic processes.
- Fibrin gels had a less pronounced effect on MFB compared to collagen gels.
