Abstract
Although hepatic myofibroblast migration plays a key role in the liver's injury response, the signal transduction pathways mediating the migration of this cell type are uncertain. Recently, we reported that lysophosphatidic acid (LPA) stimulates the migration of hepatic myofibroblasts. The goal of this study was to test the hypothesis that rho and p38 MAP kinase signaling pathways mediate LPA-stimulated hepatic myofibroblast migration. We measured migration, myosin regulatory light chain and p38 MAP kinase phosphorylation, and contractile force generation by human hepatic myofibroblasts. LPA stimulated migration in a dose-dependent and saturable manner that was partially blocked by Y-27632, a rhoassociated kinase inhibitor, as well as by SB-202190, a p38 MAP kinase inhibitor. LPA also induced myosin regulatory light chain phosphorylation and contractile force generation in a Y-27632 dependent, and SB-202190 independent fashion. Moreover, LPA stimulated a dose-dependent and saturable phosphorylation of p38 MAP kinase, which was not altered by Y-27632 or C3 transferase, a rho inactivator. These novel results suggest that LPA stimulates hepatic myofibroblast migration via distinct pathways that signal through rho and p38 MAP kinase.
Similar content being viewed by others
References
Ai S, Kuzuya M, Koike T, Asai T, Kanda S, Maeda K, Shibata T, Iguchi A. Rho-Rho kinase is involved in smooth muscle cell migration through myosin light chain phosphorylation-dependent and independent pathways. Atherosclerosis 155:321–327;2001.
Ashida N, Arai H, Yamasaki M, Kita T. Distinct signaling pathways for MCP-1-dependent integrin activation and chemotaxis. J Biol Chem 276:16555–16560;2001.
Beltman J, Erickson JR, Martin GA, Lyons JF, Cook SJ. C3 toxin activates the stress signaling pathways, JNK and p38, but antagonizes the activation of AP-1 in rat-1 cells. J Biol Chem 274:3772–3780;1999.
Davies SP, Reddy H, Caivano M, Cohen P. Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J 351:95–105;2000.
Eichholtz T, Jalink K, Fahrenfort I, Moolenaar WH. The bioactive phospholipid lysophosphatidic acid is released from activated platelets. Biochem J 291 (pt 3):677–680;1993.
Fishman DA, Liu Y, Ellerbroek SM, Stack MS. Lysophosphatidic acid promotes matrix metalloproteinase (MMP) activation and MMP-dependent invasion in ovarian cancer cells. Cancer Res 61:3194–3199;2001.
Graler MH, Goetzl EJ. Lysophospholipids and their G protein-coupled receptors in inflammation and immunity. Biochim Biophys Acta 1582:168–174;2002.
Hall A. Small GTP-binding proteins and the regulation of the actin cytoskeleton. Annu Rev Cell Biol 10:31–54;1994.
Hedges JC, Dechert MA, Yamboliev IA, Martin JL, Hickey E, Weber LA, Gerthoffer WT. A role for p38(MAPK)/HSP27 pathway in smooth muscle cell migration. J Biol Chem 274:24211–24219;1999.
Horwitz AR, Parsons JT. Cell migration-movin'on. Science 286:1102–1103;1999.
Ikeda H, Yatomi Y, Yanase M, Satoh H, Nishihara A, Kawabata M, Fujiwara K. Effects of lysophosphatidic acid on proliferation of stellate cells and hepatocytes in culture. Biochem Biophys Res Commun 248:436–440;1998.
Knittel T, Kobold D, Piscaglia F, Saile B, Neubauer K, Mehde M, Timpl R, Ramadori G. Localization of liver myofibroblasts and hepatic stellate cells in normal and diseased rat livers: Distinct roles of (myo-)fibroblast subpopulations in hepatic tissue repair. Histochem Cell Biol 112:387–401;1999.
Kolodney MS, Thimgan MS, Honda HM, Tsai G, Yee HF, Jr. Ca2+-independent myosin II phosphorylation and contraction in chicken embryo fibroblasts. J Physiol 515:87–92;1999.
Lauffenburger DA, Horwitz AF. Cell migration: A physically integrated molecular process. Cell 84:359–369;1996.
Lee H, Goetzl EJ, An S. Lysophosphatidic acid and sphingosine 1-phosphate stimulate endothelial cell wound healing. Am J Physiol Cell Physiol 278:C612–618;2000.
Lemieux P, Oesterreich S, Lawrence JA, Steeg PS, Hilsenbeck SG, Harvey JM, Fuqua SA. The small heat shock protein hsp27 increases invasiveness but decreases motility of breast cancer cells. Invasion Metastasis 17:113–123;1997.
Manning TJ, Jr., Parker JC, Sontheimer H. Role of lysophosphatidic acid and rho in glioma cell motility. Cell Motil Cytoskeleton 45:185–199;2000.
Marinissen MJ, Chiariello M, Gutkind JS. Regulation of gene expression by the small GTPase Rho through the ERK6 (p38 gamma) MAP kinase pathway. Genes Dev 15:535–553;2001.
Matsumoto T, Yokote K, Tamura K, Takemoto M, Ueno H, Saito Y, Mori S. Plateletderived growth factor activates p38 mitogenactivated protein kinase through a Ras-dependent pathway that is important for actin reorganization and cell migration. J Biol Chem 274:13954–13960;1999.
Nobes CD, Hall A. Rho GTPases control polarity, protrusion, and adhesion during cell movement. J Cell Biol 144:1235–1244;1999.
Powell DW, Mifflin RC, Valentich JD, Crowe SE, Saada JI, West AB. Myofibroblasts. I. Paracrine cells important in health and disease. Am J Physiol 277:C1-C19;1999.
Reiser CO, Lanz T, Hofmann F, Hofer G, Rupprecht HD, Goppelt-Struebe M. Lysophosphatidic acid-mediated signal-transduction pathways involved in the induction of the early-response genes prostaglandin G/H synthase-2 and Egr-1: A critical role for the mitogen-activated protein kinase p38 and for Rho proteins. Biochem J 330(pt 3):1107–1114;1998.
Ridley AJ. Rho GTPases and cell migration. J Cell Sci 114:2713–2722;2001.
Rousseau S, Houle F, Landry J, Huot J. p38 MAP kinase activation by vascular endothelial growth factor mediates actin reorganization and cell migration in human endothelial cells. Oncogene 15:2169–2177;1997.
Somlyo AP, Somlyo AV. Signal transduction by G-proteins, rho-kinase and protein phosphatase to smooth muscle and non-muscle myosin II. J Physiol 522(pt 2):177–185;2000.
Suidan HS, Nobes CD, Hall A, Monard D. Astrocyte spreading in response to thrombin and lysophosphatidic acid is dependent on the rho GTPase. Glia 21:244–252;1997.
Takuwa Y. Subtype-specific differential regulation of Rho family G proteins and cell migration by the Edg family sphingosine-1-phosphate receptors. Biochim Biophys Acta 1582:112–120;2002.
Tangkijvanich P, Melton AC, Chitapanarux T, Han J, Yee HF. Platelet-derived growth factor-BB and lysophosphatidic acid distinctly regulate hepatic myofibroblast migration through focal adhesion kinase. Exp Cell Res 281:140–147;2002.
Tangkijvanich P, Santiskulvong C, Melton AC, Rozengurt E, Yee HF, Jr. p38 MAP kinase mediates platelet-derived growth factor-stimulated migration of hepatic myofibroblasts. J Cell Physiol 191:351–361;2002.
Tangkijvanich P, Tam SP, Yee HF, Jr. Woundinduced migration of rat hepatic stellate cells is modulated by endothelin-1 through rho-kinase-mediated alterations in the acto-myosin cytoskeleton. Hepatology 33:74–80;2001.
Thimgan MS, Yee HF, Jr. Quantitation of rat hepatic stellate cell contraction: Stellate cells' contribution to sinusoidal resistance. Am J Physiol 277:G137–143;1999.
Tu LC, Chou CK, Chen HC, Yeh SF. Protein kinase C-mediated tyrosine phosphorylation of paxillin and focal adhesion kinase requires cytoskeletal integrity and is uncoupled to mitogen-activated protein kinase activation in human hepatoma cells. J Biomed Sci 8:184–190;2001.
Win KM, Charlotte F, Mallat A, Cherqui D, Martin N, Mavier P, Preaux AM, Dhumeaux D, Rosenbaum J. Mitogenic effect of transforming growth factor-beta 1 on human Ito cells in culture: Evidence for mediation by endogenous platelet-derived growth factor. Hepatology 18:137–145;1993.
Wittmann T, Waterman-Storer CM. Cell motility: Can Rho GTPases and microtubules point the way? J Cell Sci 114:3795–3803;2001.
Yanase M, Ikeda H, Matsui A, Maekawa H, Noiri E, Tomiya T, Arai M, Yano T, Shibata M, Ikeba M, Fujiwara K, Rojkind M, Ogata M. PatLysophosphatidic acid enhances collagen gel contraction by hepatic stellate cells: association with rho-kinase. Biochem Biophys Res Commun 277:72–78;2000.
Yee HF, Jr. Rho directs activation-associated changes in rat hepatic stellate cell morphology via regulation of the actin cytoskeleton. Hepatology 28:843–850;1998.
Yee HF, Jr., Melton AC, Tran BN. RhoA/rho-associated kinase mediates fibroblast contractile force generation. Biochem Biophys Res Commun 280:1340–1345;2001.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Tangkijvanich, P., Melton, A.C., Santiskulvong, C. et al. Rho and p38 MAP kinase signaling pathways mediate LPA-stimulated hepatic myofibroblast migration. J Biomed Sci 10, 352–358 (2003). https://doi.org/10.1007/BF02256455
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02256455