Abstract
To elucidate the role played by hepatic sinusoidal cells in the regulation of the circulatory status in the liver, the effect of endothelins (ETs) on primarycultured stellate cells was examined. Kinetic analysis with125I-labeled ET-1 revealed that stellate cells have ET receptors with a Kd value of 141 pM and a Bmax of 12.3 fmol/105 cells. ET-1,-2, and-3 dose-dependently increased inositol monophosphate (InsP) levels in stellate cells with an EC50 of 0.53, 1.63, and 1.88 nM, respectively. Binding of125I-labeled ET-1 to stellate cells and the ET-enhanced InsP formation were suppressed by preincubating the cells with 10 nM of unlabeled ET-1 or ET-3 for more than 3 h, indicating down-regulation and desensitization of ET receptors by homologous ligands. Binding of ETs to surface receptors induced a marked contraction of stellate cells. Stellate cells rapidly reacted to ETs, as detected by the flexible silicone-rubber-membrane method; 78%, 73%, and 58% of the stellate cells contracted 2.5 min after the addition of 10 nM of ET-1, ET-2, or ET-3, respectively. On the other hand, ETs also triggered a long-lasting contraction of the cells, as revealed with hydrated collagen gels. The ET-induced contraction of stellate cells decreased the diameter of the collagen lattice by about 60%, and this action was inhibited either by cytochalasin B or by H-7, a protein kinase C inhibitor. These and other results suggest that ETs induced cell contraction by some mechanism that involved protein kinase C.
Similar content being viewed by others
References
Yanagisawa M, Kurihara H, Kimura S, et al. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 1988;332:411–415.
Sakurai T, Yanagisawa M, Takuwa Y, et al. Cloning of a cDNA encoding a non-isopeptide selective subtype of the endothelin receptor. Nature 1990;348:732–735.
Arai H, Hori S, Aramori I, et al. Cloning and expression of a cDNA encoding an endothelin receptor. Nature 1990;348: 730–732.
Badr KF, Murray JJ, Breyer MD, et al. Mesangial cell, glomerular and renal vascular responses to endothelin in the rat kidney. Elucidation of signal transduction pathway. J Clin Invest 1989;83:336–342.
Furuya S, Naruse S, Nakayama T, et al. Localization of [125I]endothelin-1 in rat tissue observed by electron-microscopic radioautography. J Cardiovasc Pharmacol 1991;17:S452-S454.
Gondo K, Ueno T, Sakamoto M, et al. The endothelin-1 binding site in rat liver tissue: Light and electron microscopic autoradiographic studies. Gastroenterology 1993;104:1745–1749.
Gal CS-L, Jouneaux C, Sanchez-Bueno A, et al. Endothelin action in rat liver. Receptor, free Ca2+ oscillation, and activation of glycogenolysis. J Clin Invest 1991;87:133–138.
Tran-Thi T-A, Kawada N, Decker K. Regulation of endothelin 1 action on the perfused rat liver. FEBS Lett. 1993;318:353–357.
Gandhi CR, Stephenson K, Olson MS. Endothelin, a potent peptide agonist in the liver. J Biol Chem 1990;265:17432–17435.
Gandhi CR, Stephenson K, Olson MS. A comparative study of endothelin- and platelet-activating factor-mediated signal transduction and prostaglandin synthesis in rat Kupffer cells. Biochem J 1992;281:485–492.
Wake K. Perisinusoidal stellate cells (fat-storing cells, interstitial cells, lipocytes), their related structure in and around the liver sinusoids, and vitamin A-storing cells in extrahepatic organs. Int Rev Cytol 1980;66:302–353.
Friedman SL, Roll FJ, Boyles J et al. Hepatic lipocytes: The principal collagen-producing cells of normal rat liver. Proc Natl Acad Sci USA 1985;82:8681–8685.
Gressner AM, Althaus M. Effect of ethanol, acetoaldehyde, and lactate on proteoglycan synthesis and proliferation of cultured rat liver fat-storing cells. Gastroenterology 1988;94:797–807.
Friedman SL, Arthur MJP. 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 1989;84:1780–1785.
Pinzani M, Gesualdo L, Sabbah GM, et al. Effect of platelet-derived growth factor and other polypeptide mitogens on DNA synthesis and growth of cultured rat liver fat-storing cells. J Clin Invest 1989;84:1786–1793.
Kawada N, Klein H, Decker K. Eicosanoid-mediated contractility of hepatic stellate cells. Biochem J 1992;285:367–371.
Kawada N, Tran-Thi T-A, Klein H, et al. The contraction of hepatic stellate (Ito) cells stimulated with vasoactive substances. Possible invovement of endothelin 1 and nitric oxide in the regulation of the sinusoidal tonus. Eur J Biochem 1993;213: 815–823.
Kawada N, Klein H, Decker K. Contraction and relaxation of hepatic stellate cells in culture. In: Knook DL, Wisse E (eds) Cells of the hepatic sinusoid. Vol. 4. Leiden: The Kupffer Cell Foundation, 1993;161–164.
Sakamoto M, Ueno T, Kin M, et al. Ito cell contraction in response to endothelin-1 and substance P. Hepatology 1993;18: 978–983.
Housset C, Rockey DC, Bissell DM. Endothelin receptors in rat liver: Lipocytes as a contractile target for endothelin 1. Proc Natl Acad Sci USA 1993;90:9266–9270.
Yokoi Y, Namihisa T, Kuroda H, et al. Immunocytochemical detection of desmin in fat-storing cells (Ito cells). Hepatology 1984;4:709–714.
Berridge MJ, Dawson RMC, Downes CP, et al. Changes in the levels of inositol phosphates after agonist-dependent hydrolysis of membrane phosphoinositides. Biochem J 1983;212:473–482.
Kelley C, D'Amore P, Hechtman HB, et al. Microvascular pericyte contractility in vitro: Comparison with other cells of the vascular wall. J Cell Biol 1987;104:483–490.
Guidry C, Grinnel F. Endotheline produced by endothelial cells promotes collagen gel contraction by fibroblasts. J Cell Biol 1991;115:873–880.
Rockey DC, Maher JJ, Jarnagin WR, et al. Inhibition of rat hepatic lipocyte activation in culture by interferon γ. Hepatology 1992;16:776–784.
Kawamoto S, Hidaka H. 1-(5-Isoquinolinesulfonyl)-2-methyl-piperazine (H-7) is a selective inhibitor of protein kinase C in rabbit platelets. Biochem Biophys Res Commun 1984;125: 258–262.
Gandhi CR, Behal RH, Harvey SAK, et al. Hepatic effect of endothelin. Receptor characterization and endothelin-induced signal transduction in hepatocytes. Biochem J 1992;287:897–904.
Reynolds EE, Mok LLS. Role of thromboxane A2/prostaglandin H2 receptor in the vasoconstrictor response of rat aorta to endothelin. J Pharmacol Exp Ther 1990;252:915–921.
Thiermermann C. Biosynthesis and interaction of endothelium-derived vasoactive mediators. Eicosanoids 1991;4:187–202.
Kuiper J, Zijlstra FJ, Kamp JAAM. Identification of prostaglandin D2 as the major eicosanoid from liver endothelial and Kupffer cells. Biochim Biophys Acta 1988;959:143–152.
Rubanyi GM, Botelho LHP. Endothelins. FASEB J 1991; 5:2713–2720.
Pastan IH, Willingham MC. Receptor-mediated endocytosis of hormones in cultured cells. Annu Rev Physiol 1981;43:239–250.
Schrader J, Tabbe U, Borries M, et al. Plasma-endothelin bei Normalpersonen und Patienten mit nephrologisch-rheumatologischen und kardiovascularen Erkrankungen. Klin Wochenschr 1990;68:774–779.
Rieder H, Ramadori G, Meyer zum Buschenfelde K-H. Sinusoidal endothelial liver cells in vitro release endothelin-augmentation by transforming growth factor β and Kupffer cell-conditioned media. Klin Wochenschr 1991;69:387–391.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kawada, N., Kuroki, T., Kobayashi, K. et al. Action of endothelins on hepatic stellate cells. J Gastroenterol 30, 731–738 (1995). https://doi.org/10.1007/BF02349639
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02349639