Summary
Hepatic perisinusoidal cells (PSCs) proliferate and are thought to be the principal source of extracellular matrix proteins during the development of liver fibrosis. We have studied the classical model of carbon tetrachloride induced liver fibrosis in order to evaluate the possible modulation of PSCs into a synthetically active and contractile cell: the myofibroblast (MF). At the ultrastructural level, this modulation was characterized by reduction of lipid vacuoles and appearance of a developed rough endoplasmic reticulum as well as of microfilament bundles. On investigating the cytoskeletal equipment of PSCs and MFs using light and electron microscopic immunohistochemistry, we found a heterogeneity of phenotypic features. While typical PSCs in normal and fibrotic livers always contained desmin, MFs expressedα-smooth muscle (SM) actin in areas of tissue injury and active fibrogenesis. Cells co-expressingα-SM actin and desmin were most prominent in the prevenular zone of the lobule (known to be vulnerable to carbon tetrachloride toxicity) and in developing fibrous septa. As demonstrated by immunogold electron microscopy, labelling of microfilament bundles byα-SM actin antibody was noted in PSCs containing lipid droplets in early stages of fibrosis; here MFs gradually accumulated and showedα-SM actin containing microfilament bundles. In scar tissue,α-SM actin expression decreased in both PSCs and myofibroblasts. Our observations support the concept of phenotypic plasticity of PSCs and confirm, at the ultrastructural level, previous suggestions of modulation of these cells into MFs in the course of liver fibrosis.
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References
Aterman K (1986) The parasinusoidal cells of the liver: a historical account. Histochem J 18:279–305
Bachem MG, Meyer D, Melchior R, Sell KM, Gressner AM (1992) Activation of rat liver perisinusoidal lipocytes by transforming growth factors derived from myofibroblastlike cells. A potential mechanism of self perpetuation in liver fibrogenesis. J Clin Invest 89:19–27
Blomhoff R, Wake K (1991) Perisinusoidal stellate cells of the liver: important roles in retinol metabolism and fibrosis. FASEB J 5:271–277
Bolen JW, Hammar SP, McNutt MA (1986) Reactive and neoplastic serosal tissue. A light microscopic, ultrastructural, and immunocytochemical study. Am J Surg Pathol 10:34–47
Czaja MJ, Weiner FR, Flanders KC, Giambrone MA, Wind R, Biempica L, Zern MA (1989) In vitro and in vivo association of transforming growth factor-β1 with hepatic fibrosis. J Cell Biol 108:2477–2482
Danscher G (1981) Localization of gold in biological tissue. A photochemical method for both light and electron microscopy. Histochemistry 71:81–89
Darby I, Skalli O, Gabbiani G (1990)α-Smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Invest 63:21–29
French SW, Miyamoto K, Wong K, Jui L, Briere L (1988) Role of the Ito cell in liver parenchymal fibrosis in rats fed alcohol and a high fat-low protein diet. Am J Pathol 132:73–85
Friedman SL, Arthur MJP (1989) Activation of cultured rat hepatic lipocytes by Kupffer cell conditioned medium. Direct enhancement of matrix synthesis and stimulation of cell proliferation via induction of platelet-derived growth factor receptors. J Clin Invest 84:1750–1755
Friedman SL, Bissell DM (1990) Hepatic fibrosis: new insights into pathogenesis. Hosp Pract 25:43–50
Friedman SL, Roll FJ, Boyles J, Bissell DM (1985) Hepatic lipocytes: the principal collagen-producing cells of normal rat liver. Proc Natl Acad Sci USA 82:8681–8685
Friedman SL, Roll JF, Boyles J, Arenson DM, Bissell DM (1989) Maintenance of differentiated phenotype of cultured rat hepatic lipocytes by basement membrane matrix. J Biol Chem 264:10756–10762
Gabbiani G, Ryan GB, Majno G (1971) Presence of modified fibroblasts in granulation tissue and their possible role in wound contracture. Experientia 27:549–550
Gabbiani G, Montesano R, Tuchweber B, Salas M, Orci L (1975) Phalloidin-induced hyperplasia of actin filaments in rat hepatocytes. Lab Invest 33:562–569
Geerts A, Lazou JM, De Bleser P, Wisse E (1991) Tissue distribution, quantitation and proliferation kinetics of fat-storing cells in carbon tetrachloride-injured rat liver. Hepatology 13:1193–1202
Gown AM (1990) The mysteries of the myofibroblast (partially) unmasked. Lab Invest 63:1–3
Gressner AM (1991)β-D-xyloside induced modulations of glycosaminoglycans, proliferation, and cytoskeletal organization of rat liver myofibroblast-like cells (transformed fat storing cells). Cell Mol Biol 37:549–564
Herbst H, Milani S, Schuppan D, Stein H (1991) Temporal and spatial patterns of proto-oncogene expression at early stages of toxic liver injury in the rat. Lab Invest 65:324–334
Irle C, Kocher O, Gabbiani G (1980) Contractility of myofibroblasts during experimental liver cirrhosis. J Submicrosc Cytol 12:209–217
Kocher O, Skalli O, Bloom WS, Gabbiani G (1984) Cytoskeleton of rat aortic smooth muscle cells. Normal conditions and experimental intimal thickening. Lab Invest 50:645–652
Kuhn C, McDonald JA (1991) The roles of the myofibroblast in idiopathic pulmonary fibrosis. Ultrastructural and immunohistochemical features of sites of active extracellular matrix synthesis. Am J Pathol 138:1257–1265
Leo MA, Lieber CS (1983) Hepatic fibrosis after long-term administration of ethanol and moderate vitamin A supplementation in the rat. Hepatology 3:1–11
Mak KM, Lieber CS (1988) Lipocytes and transitional cells in alcoholic liver disease: a morphometric study. Hepatolgy 8:1027–1033
Mak KM, Leo MA, Lieber CS (1984) Alcoholic liver injury in baboons: transformation of lipocytes to transitional cells. Gastroenterology 87:188–200
Martinez-Hernandez A (1985) The hepatic extracellular matrix. II. Electron immunohistochemical studies in rats with CCl4-induced cirrhosis. Lab Invest 53:166–186
Matsuoka M, Pham NT, Tsukamoto H (1989) Differential effects of interleukin-1α, tumor necrosis factorα, and transforming growth factorβ1 on cell proliferation and collagen formation by cultured fat-storing cells. Liver 9:71–78
Matsuoka M, Zhang MY, Tsukamoto H (1990) Sensitization of hepatic lipocytes by high-fat diet to stimulatory effects of Kupffer cell-derived factors: implication in alcoholic liver fibrogenesis. Hepatology 11:173–182
Miao S, Bao-En W, Annoni G, Degli Esposti S, Biempica I, Zern MA (1990) Two rat models of hepatic fibrosis. A morphologic and molecular comparison. Lab Invest 63:467–475
Milani S, Herbst H, Schuppan D, Hahn EG, Stein H (1989) In situ hybridization for procollagen types I, III and IV mRNA in normal and fibrotic rat liver: evidence for predominant expression in nonparenchymal liver cells. Hepatology 10:84–92
Milani S, Herbst H, Schuppan D, Kim KY, Riecken EO, Stein H (1990) Procollagen expression by nonparenchymal rat liver cells in experimental biliary fibrosis. Gastroenterology 98:175–184
Nakano M, Lieber CS (1982) Ultrastructure of initial stages of perivenular fibrosis in alcohol-fed baboons. Am J Pathol 106:145–155
Nehls V, Drenckhahn D (1991) Heterogeneity of microvascular pericytes for smooth muscle type alpha-actin. J Cell Biol 113:147–154
Ogawa K, Suzuki JI, Mukai H, Mori M (1986) Sequential changes of extracellular matrix and proliferation of Ito cells with enhanced expression of desmin and actin in focal hepatic injury. Am J Pathol 125:611–619
Pinzani M, Gesualdo L, Sabbah GM, Abboud HE (1989) Effects of platelet-derived growth factor and other polypeptide mitogens on DNA synthesis and growth of cultured rat liver fatstoring cells. J Clin Invest 84:1786–1793
Ramadori G (1991) The stellate cell (Ito-cell, fat-storing cell, lipocyte, perisinusoidal cell) of the liver. New insights into pathophysiology of an intriguing cell. Virchows Arch [B] 61:147–158
Ramadori G, Veit T, Schwögler S, Kienes HP, Knittel T, Rieder H, Meyer zum Büschenfelde KH (1990) Expression of the gene of theα-smooth muscle-actin isoform in rat liver and in rat fat-storing (Ito) cells. Virchows Arch [B] 59:349–357
Rockey DC, Boyles JK, Gabbiani G, Friedman SL (1992) Rat hepatic lipocytes express smooth muscle actin upon activation in vivo and in culture. J Submicrosc Cytol Pathol 24:193–203
Ronnov-Jessen L, Van Deurs B, Celis JE, Petersen OW (1990) Smooth muscle differentiation in cultured human breast gland stromal cells. Lab Invest 63:532–543
Rubbia-Brandt L, Sappino AP, Gabbiani G (1991) Locally applied GM-CSF induces the accumulation ofα-smooth muscle actin containing myofibroblasts. Virchows Arch [B] 60:73–82
Rungger-Brändle E, Gabbiani G (1983) The role of cytoskeletal and cytocontractile elements in pathological processes. Am J Pathol 110:359–392
Sappino AP, Schürch W, Gabbiani G (1990) Differentiation repertoire of fibroblastic cells: expression of cytoskeletal proteins as marker of phenotypic modulations. Lab Invest 63:144–161
Schmitt-Gräff A, Skalli O, Gabbiani G (1989)α-Smooth muscle actin is expressed in a subset of bone marrow stromal cells in normal and pathological conditions. Virchows Arch [B] 57:291–302
Schmitt-Gräff A, Pau H, Spahr R, Piper HM, Skalli O, Gabbiani G (1990) Appearance of alpha-smooth muscle actin in human eye lens cells of anterior capsular cataract and in cultured bovine lens-forming cells. Differentiation 43:115–122
Schmitt-Gräff A, Krüger S, Bochard F, Gabbiani G, Denk H (1991) Modulation of alpha smooth muscle actin and desmin expression in perisinusoidal cells of normal and diseased human livers. Am J Pathol 138:1233–1242
Self DA, Schroeder PC, Gown AM (1988) Hamster thecal cells express muscle characteristics. Biol Reprod 39:119–130
Skalli O, Gabbiani G (1988) The biology of the myofibroblast: relationship to wound contraction and fibrocontractive disease. In: Clark RAF, Henson PM (eds) The molecular and cellular biology of wound repair. Plenum, New York, pp 373–402
Skalli O, Ropraz P, Trzeciak A, Benzonana G, Gillessen D, Gabbiani G (1986) A monoclonal antibody againstα-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol 103:2787–2796
Suematsu M, Kato S, Ishii H, Asako H, Yanagisawa T, Suzuki H, Oshio C, Tsuchiya M (1991) Intralobular heterogeneity of carbon tetrachloride-induced oxidative stress in perfused rat liver visualized by digital imaging fluorescence microscopy. Lab Invest 64:167–173
Takase S, Leo MA, Nouchi T, Lieber CS (1988) Desmin distinguishes cultured fat-storing cells from myofibroblasts, smooth muscle cells and fibroblasts in the rat. J Hepatol 6:267–276
Tanaka Y, Nouchi T, Yamane M, Yamaoka K, Irie T, Sato C, Marumo F (1990) Expression of alpha-smooth muscle actin in lipocytes in hepatic fibrosis: an evidence for modulation in lipocytes (abstract). Hepatology 12:953
Tsutsumi M, Takada A, Takase S (1987) Characterization of desmin-positive rat liver sinusoidal cells. Hepatology 7:277–284
Weiner FR, Giambrone MA, Czaja MJ, Shah A, Annoni G, Takahashi S, Eghbali M, Zern MA (1990) Ito-cell gene expression and collagen regulation. Hepatology 11:111–117
Yokoi Y, Namihisa T, Kuroda H, Komatsu I, Miyazaki A, Watanabe S, Usui K (1984) Immunocytochemical detection of desmin in fat-storing cells (Ito cells). Hepatology 4:709–714
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Schmitt-Gräff, A., Chakroun, G. & Gabbiani, G. Modulation of perisinusoidal cell cytoskeletal features during experimental hepatic fibrosis. Vichows Archiv A Pathol Anat 422, 99–107 (1993). https://doi.org/10.1007/BF01607161
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DOI: https://doi.org/10.1007/BF01607161