The Myofibroblastic Conversion of Peribiliary Fibrogenic Cells Distinct from Hepatic Stellate Cells Is Stimulated by Platelet-Derived Growth Factor During Liver Fibrogenesis

The origin of myofibroblasts and the factors promoting their differentiation during liver fibrogenesis remain uncertain. During biliary-type fibrogenesis, the proliferation and chemoattraction of hepatic stellate cells (HSC) toward bile ducts is mediated by platelet-derived growth factor (PDGF), while myofibroblastic conversion of peribiliary cells distinct from HSC also occurs. We herein examined the phenotype of these peribiliary myofibroblasts as compared with myofibroblastic HSC and tested whether their differentiation was affected by PDGF. Biliary-type liver fibrogenesis was induced by common bile duct ligation in rats. After 48 hours, periductular fibrosis in portal tracts colocalized with smooth muscle α-actin–immunoreactive myofibroblasts, the majority of which were desmin negative. Simultaneously, in sinusoids, desmin immunoreactivity was induced in a large number of HSC, which were smooth muscle α-actin negative. Cultures of peribiliary myofibroblasts were expanded from isolated bile duct segments and compared with myofibroblastic HSC. Peribiliary myofibroblasts outgrowing from bile duct segments expressed smooth muscle α-actin, α1 (I) collagen mRNA, and PDGF receptor-β subunit. Desmin immunoreactivity gradually decreased in cultured peribiliary myofibroblasts, contrasting with constant labeling of all myofibroblastic HSC. In addition, IL-6 expression in peribiliary myofibroblasts was up to 100-fold lower than in myofibroblastic HSC, whereas the expression of the complement-activating protease P100 in both cell types showed little difference and that of the extracellular matrix component fibulin 2 was similar. The expression of smooth muscle α-actin protein in cultured peribiliary myofibroblasts was stimulated by PDGF-BB and inhibited by STI571, a PDGF receptor tyrosine kinase inhibitor, whereas in bile duct–ligated rats, the administration of STI571 caused a significant decrease in peribiliary smooth muscle α-actin immunoreactivity, and to a lesser extent, a decrease in peribiliary fibrosis. These results indicate that peribiliary cells distinct from HSC undergo a PDGF-mediated conversion into myofibroblasts expressing IL-6 at lower levels than myofibroblastic HSC and contribute to the initial formation of biliary-type liver fibrosis.

[1]  F. Marumo,et al.  Phenotypic change in portal fibroblasts in biliary fibrosis. , 2008, Liver.

[2]  A. Desmoulière,et al.  Cellular Retinol-Binding Protein-1 Expression and Modulation during In Vivo and In Vitro Myofibroblastic Differentiation of Rat Hepatic Stellate Cells and Portal Fibroblasts , 2002, Laboratory Investigation.

[3]  D. Wendum,et al.  Hypoxia‐induced VEGF and collagen I expressions are associated with angiogenesis and fibrogenesis in experimental cirrhosis , 2002, Hepatology.

[4]  D. Wendum,et al.  Hepatic Stellate Cell Proliferation is an Early Platelet-Derived Growth Factor-Mediated Cellular Event in Rat Cholestatic Liver Injury , 2001, Laboratory Investigation.

[5]  D. Zhou,et al.  Transcription of the Schizosaccharomyces pombe U2 gene in vivo and in vitro is directed by two essential promoter elements. , 2001, Nucleic acids research.

[6]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[7]  T. Roskams,et al.  Hepatic stellate cell/myofibroblast subpopulations in fibrotic human and rat livers. , 2001, Journal of hepatology.

[8]  T. Saibara,et al.  Appearance of denuded hepatic stellate cells and their subsequent myofibroblast-like transformation during the early stage of biliary fibrosis in the rat , 2000, Medical Electron Microscopy.

[9]  B. Druker,et al.  Abl protein-tyrosine kinase inhibitor STI571 inhibits in vitro signal transduction mediated by c-kit and platelet-derived growth factor receptors. , 2000, The Journal of pharmacology and experimental therapeutics.

[10]  R. Hultcrantz,et al.  PDGF-Mediated Chemoattraction of Hepatic Stellate Cells by Bile Duct Segments in Cholestatic Liver Injury , 2000, Laboratory Investigation.

[11]  S. Friedman.  Molecular Regulation of Hepatic Fibrosis, an Integrated Cellular Response to Tissue Injury* , 2000, The Journal of Biological Chemistry.

[12]  R. Timpl,et al.  Localization of liver myofibroblasts and hepatic stellate cells in normal and diseased rat livers: distinct roles of (myo-)fibroblast subpopulations in hepatic tissue repair , 1999, Histochemistry and Cell Biology.

[13]  F. Piscaglia,et al.  Rat liver myofibroblasts and hepatic stellate cells: different cell populations of the fibroblast lineage with fibrogenic potential. , 1999, Gastroenterology.

[14]  H. Herbst,et al.  Expression of platelet-derived growth factor in newly formed cholangiocytes during experimental biliary fibrosis in rats. , 1999, Journal of hepatology.

[15]  N. Kawada,et al.  In Vitro Migratory potential of rat quiescent hepatic stellate cells and its augmentation by cell activation , 1999, Hepatology.

[16]  L. Hammarström,et al.  Expression of Collagen α1(I) mRNA Variants during Tooth and Bone Formation in the Rat , 1999, Journal of dental research.

[17]  T. B. Morrison,et al.  Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification. , 1998, BioTechniques.

[18]  H. Kurz,et al.  Platelet-derived growth factor-B induces transformation of fibrocytes into spindle-shaped myofibroblasts in vivo , 1998, Histochemistry and Cell Biology.

[19]  D. Rockey,et al.  Cellular localization of endothelin‐1 and increased production in liver injury in the rat: Potential for autocrine and paracrine effects on stellate cells , 1998, Hepatology.

[20]  E. Furth,et al.  Liver Failure and Defective Hepatocyte Regeneration in Interleukin-6-Deficient Mice , 1996, Science.

[21]  R. Demir,et al.  Placental villous stroma as a model system for myofibroblast differentiation , 1996, Histochemistry and Cell Biology.

[22]  E. Wisse,et al.  Experimental biliary fibrosis correlates with increased numbers of fat-storing and Kupffer cells, and portal endotoxemia. , 1995, Journal of hepatology.

[23]  M. Pinzani.  Induction of β-platelet-derived growth factor receptor in rat hepatic lipocytes during cellular activation in vivo and in culture , 1995 .

[24]  R. Chamuleau,et al.  Interleukin-6 production by human liver (myo)fibroblasts in culture. Evidence for a regulatory role of LPS, IL-lβ and TNFα , 1995 .

[25]  J. Boyer,et al.  Isolation of small polarized bile duct units. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[26]  P. van Eyken,et al.  Ductular reaction in the liver. , 1995, Pathology, research and practice.

[27]  S. Friedman,et al.  Induction of beta-platelet-derived growth factor receptor in rat hepatic lipocytes during cellular activation in vivo and in culture. , 1994, The Journal of clinical investigation.

[28]  A. Burt,et al.  In vivo responses of macrophages and perisinusoidal cells to cholestatic liver injury. , 1993, The American journal of pathology.

[29]  E. Wisse,et al.  Tissue distribution, quantitation and proliferation kinetics of fat‐storing cells in carbon tetrachloride–injured rat liver , 1991, Hepatology.

[30]  R. F. McGuire,et al.  Extracellular matrix gene expression increases preferentially in rat lipocytes and sinusoidal endothelial cells during hepatic fibrosis in vivo. , 1990, The Journal of clinical investigation.

[31]  L. Gesualdo,et al.  Effects of platelet-derived growth factor and other polypeptide mitogens on DNA synthesis and growth of cultured rat liver fat-storing cells. , 1989, The Journal of clinical investigation.

[32]  F. Dautry,et al.  Northern blot normalization with a 28S rRNA oligonucleotide probe. , 1989, Nucleic acids research.

[33]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[34]  S. Friedman,et al.  Isolation and culture of hepatic lipocytes, Kupffer cells, and sinusoidal endothelial cells by density gradient centrifugation with Stractan. , 1987, Analytical biochemistry.

[35]  W. Webb,et al.  Fluorescent erythrosin B is preferable to trypan blue as a vital exclusion dye for mammalian cells in monolayer culture. , 1984, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[36]  J. Kountouras,et al.  Prolonged bile duct obstruction: a new experimental model for cirrhosis in the rat. , 1984, British journal of experimental pathology.

[37]  D. Knook,et al.  Purified Rat Liver Fat‐Storing Cells in Culture Divide and Contain Collagen , 1984, Hepatology.

[38]  D. Knook,et al.  Fat-storing cells of the rat liver. Their isolation and purification. , 1982, Experimental cell research.

[39]  A. Desmoulière,et al.  Heterogeneity of myofibroblast phenotypic features: an example of fibroblastic cell plasticity , 2004, Virchows Archiv.

[40]  T. Meyer,et al.  Inhibition of the Abl protein-tyrosine kinase in vitro and in vivo by a 2-phenylaminopyrimidine derivative. , 1996, Cancer research.

[41]  L. Rubbia‐Brandt,et al.  Proliferation and phenotypic modulation of portal fibroblasts in the early stages of cholestatic fibrosis in the rat. , 1996, Laboratory investigation; a journal of technical methods and pathology.

[42]  D. Schuppan,et al.  Cellular sources of extracellular matrix proteins in normal and fibrotic liver. Studies of gene expression by in situ hybridization. , 1995, Journal of hepatology.

[43]  R. Chamuleau,et al.  Interleukin-6 production by human liver (myo)fibroblasts in culture. Evidence for a regulatory role of LPS, IL-1 beta and TNF alpha. , 1995, Journal of hepatology.

[44]  Y. Horsmans,et al.  Liver injury related to amoxycillin-clavulanic acid: interlobular bile-duct lesions and extrahepatic manifestations. , 1995, Journal of hepatology.

[45]  C. Lieber,et al.  Desmin distinguishes cultured fat-storing cells from myofibroblasts, smooth muscle cells and fibroblasts in the rat. , 1988, Journal of hepatology.