REGULATORY ROLE OF EXTRACELLULAR MATRIX COMPONENTS IN EXPRESSION OF MATRIX METALLOPROTELNASES IN CULTURED HEPATIC STELLATE CELLS

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Hepatic stellate cells (HSC) were found to change their morphology and function including production of matrix metalloproteinases (MMPs) in response to extracellular matrix (ECM) components used as a substratum in culture. We examined in this study the regulatory role of ECM components in expression of MMPs and tissue inhibitor of metalloproteinase (TIMP) in rat cultured HSCs. HSCs were isolated by the collagenase perfusion method, followed by centrifugation on the Percoll density gradient, and cultured in polystyrene dish, on type I collagen-coated surface, on type I collagen gel, or on Matrigel containing basement membrane components, respectively. When cultured on type I collagen gel, HSCs showed the asteroid cell shape with many elongated processes, and displayed MMP-1 activity in the culture medium, as detected by in situ zymography. Expression of MMP-1 protein and mRNA were also enhanced in HSCs cultured on type I collagen gel, as observed by immunofluorescence staining and RT-PCR analysis. Active form of MMP-2 was detected by gelatin zymography in the conditioned medium of HSCs cultured on type I collagen gel, whereas proMMP-2 but not active form of MMP-2 was detected when HSCs were cultured on polystyrene surface, on type I collagen-coated surface, or on Matrigel. Increased MMP-2 mRNA was also detected by RT-PCR in HSCs cultured on type I collagen gel, as compared to the HSCs cultured on polystyrene surface, on type I collagen-coated surface, or on Matiigel. MT1-MMP protein and mRNA expression, as examined by immunoblotting and RT-PCR analysis, were greatly enhanced in HSCs cultured on type I collagen gel, suggesting an involvement of MT1-MMP in proMMP-2 activation. TIMP-2 mRNA expression was also detected by RT-PCR analysis in HSCs cultured on type I collagen-coated surface or on type I collagen gel, although it remains unclear whether TIMP-2 is involved in proMMP-2 activation or in suppression of active form of MMP-2 in HSC culture. These results indicate that the expression of MMPs and TIMP-2 in cultured HSCs is regulated by ECM components used as substratum, suggesting an important role of HSCs in the remodelling of ECM in perisinusoidal spaces of the liver tissue.
日本結合組織学会の論文

REGULATORY ROLE OF EXTRACELLULAR MATRIX COMPONENTS IN EXPRESSION OF MATRIX METALLOPROTELNASES IN CULTURED HEPATIC STELLATE CELLS

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