p38 Kinase Is a Negative Regulator of Angiotensin II Signal Transduction in Vascular Smooth Muscle Cells

Effects on Na⁺/H⁺ Exchange and ERK1/2

From the Department of Medicine, University of Washington (M.K., E.T., T.E.P., J.A.), Seattle, and the Department of Immunology, The Scripps Research Institute (J.H., R.U.), La Jolla, Calif.

Correspondence to Bradford C. Berk, University of Rochester Medical Center, Cardiology Unit, Box 679, Rochester, NY 14642. E-mail Bradford_Berk{at}urmc.rochester.edu

Abstract—Activation of the Na⁺/H⁺ exchanger isoform-1 (NHE-1) by angiotensin II is an early signal transduction event that may regulate vascular smooth muscle cell (VSMC) growth and migration. Many signal transduction events stimulated by angiotensin II are mediated by the mitogen-activated protein (MAP) kinases. To define their roles in angiotensin II–mediated NHE-1 activity, VSMCs were treated with angiotensin II and the activities of p38, c-Jun N-terminal kinase (JNK), and extracellular signal–regulated kinases 1 and 2 (ERK1/2) were measured. Angiotensin II rapidly (peak, 5 minutes) activated p38 and ERK1/2, whereas JNK was activated more slowly (peak, 30 minutes). Because angiotensin II stimulated Na⁺/H⁺ exchange within 5 minutes, the effects of p38 and ERK1/2 antagonists on Na⁺/H⁺ exchange were studied. The MEK-1 inhibitor PD98059 decreased ERK1/2 activity and Na⁺/H⁺ exchange stimulated by angiotensin II. In contrast, the specific p38 antagonist SKF-86002 increased Na⁺/H⁺ exchange. Two mechanisms were identified that may mediate the effects of p38 and SKF-86002 on angiotensin II–stimulated Na⁺/H⁺ exchange. First, angiotensin II activation of ERK1/2 was increased 1.5- to 2.5-fold (depending on assay technique) in the presence of SKF-86002, demonstrating that p38 negatively regulates ERK1/2. Second, the ability of angiotensin II–stimulated MAP kinases to phosphorylate a glutathione S-transferase fusion protein containing amino acids 625 to 747 of NHE-1 in vitro was analyzed. The relative activities of endogenous immunoprecipitated p38, ERK1/2, and JNK were 1.0, 2.0, and 0.05 versus control, respectively suggesting that p38 and ERK1/2, but not JNK, may phosphorylate NHE-1 in VSMC. These data indicate important roles for p38 and ERK1/2 in angiotensin II–mediated regulation of the Na⁺/H⁺ exchanger in VSMC.

Key Words: mitogen-activated protein kinase • Na⁺/H⁺ exchange • angiotensin II • vascular smooth muscle

This article has been cited by other articles:

				J. Ino, C. Kojima, M. Osaka, K. Nitta, and M. Yoshida Dynamic Observation of Mechanically-Injured Mouse Femoral Artery Reveals an Antiinflammatory Effect of Renin Inhibitor Arterioscler Thromb Vasc Biol, November 1, 2009; 29(11): 1858 - 1863. [Abstract] [Full Text] [PDF]

				Y. H. HAN, H. J. MOON, B. R. YOU, S. Z. KIM, S. H. KIM, and W. H. PARK The Effect of MAPK Inhibitors on Arsenic Trioxide-treated Calu-6 Lung Cells in Relation to Cell Death, ROS and GSH Levels Anticancer Res, October 1, 2009; 29(10): 3837 - 3844. [Abstract] [Full Text] [PDF]

				L. Yu, D. A. Quinn, H. G. Garg, and C. A. Hales Deficiency of the NHE1 Gene Prevents Hypoxia-induced Pulmonary Hypertension and Vascular Remodeling Am. J. Respir. Crit. Care Med., June 1, 2008; 177(11): 1276 - 1284. [Abstract] [Full Text] [PDF]

				B. C. Berk Atheroprotective Signaling Mechanisms Activated by Steady Laminar Flow in Endothelial Cells Circulation, February 26, 2008; 117(8): 1082 - 1089. [Full Text] [PDF]

				G.-X. Zhang, X.-M. Lu, S. Kimura, and A. Nishiyama Role of mitochondria in angiotensin II-induced reactive oxygen species and mitogen-activated protein kinase activation Cardiovasc Res, November 1, 2007; 76(2): 204 - 212. [Abstract] [Full Text] [PDF]

				S. Aker, A. K Snabaitis, I. Konietzka, A. van de Sand, K. Bongler, M. Avkiran, G. Heusch, and R. Schulz Inhibition of the Na+/H+ exchanger attenuates the deterioration of ventricular function during pacing-induced heart failure in rabbits Cardiovasc Res, August 1, 2004; 63(2): 273 - 282. [Abstract] [Full Text] [PDF]

				A. D. Schecter, A. B. Berman, L. Yi, H. Ma, C. M. Daly, K. Soejima, B. J. Rollins, I. F. Charo, and M. B. Taubman MCP-1-dependent signaling in CCR2-/- aortic smooth muscle cells J. Leukoc. Biol., June 1, 2004; 75(6): 1079 - 1085. [Abstract] [Full Text] [PDF]

				J. A. Aguirre-Ghiso, Y. Estrada, D. Liu, and L. Ossowski ERKMAPK Activity as a Determinant of Tumor Growth and Dormancy; Regulation by p38SAPK Cancer Res., April 1, 2003; 63(7): 1684 - 1695. [Abstract] [Full Text] [PDF]

				S. Kalyankrishna and K. U. Malik Norepinephrine-Induced Stimulation of p38 Mitogen-Activated Protein Kinase Is Mediated by Arachidonic Acid Metabolites Generated by Activation of Cytosolic Phospholipase A2 in Vascular Smooth Muscle Cells J. Pharmacol. Exp. Ther., February 1, 2003; 304(2): 761 - 772. [Abstract] [Full Text] [PDF]

				E. C. Rothstein, K. L. Byron, R. E. Reed, L. Fliegel, and P. A. Lucchesi H2O2-induced Ca2+ overload in NRVM involves ERK1/2 MAP kinases: role for an NHE-1-dependent pathway Am J Physiol Heart Circ Physiol, August 1, 2002; 283(2): H598 - H605. [Abstract] [Full Text] [PDF]

				B. A. Watts III and D. W. Good ERK mediates inhibition of Na+/H+ exchange and HCO3- absorption by nerve growth factor in MTAL Am J Physiol Renal Physiol, June 1, 2002; 282(6): F1056 - F1063. [Abstract] [Full Text] [PDF]

				E. Tsiani, P. Lekas, I. G. Fantus, J. Dlugosz, and C. Whiteside High glucose-enhanced activation of mesangial cell p38 MAPK by ET-1, ANG II, and platelet-derived growth factor Am J Physiol Endocrinol Metab, January 1, 2002; 282(1): E161 - E169. [Abstract] [Full Text] [PDF]

				G. G. Goss, L. Jiang, D. H. Vandorpe, D. Kieller, M. N. Chernova, M. Robertson, and S. L. Alper Role of JNK in hypertonic activation of Cl--dependent Na+/H+ exchange in Xenopus oocytes Am J Physiol Cell Physiol, December 1, 2001; 281(6): C1978 - C1990. [Abstract] [Full Text] [PDF]

				A. R. Khaled, A. N. Moor, A. Li, K. Kim, D. K. Ferris, K. Muegge, R. J. Fisher, L. Fliegel, and S. K. Durum Trophic Factor Withdrawal: p38 Mitogen-Activated Protein Kinase Activates NHE1, Which Induces Intracellular Alkalinization Mol. Cell. Biol., November 15, 2001; 21(22): 7545 - 7557. [Abstract] [Full Text] [PDF]

				S. Wei, E. C. Rothstein, L. Fliegel, L. J. Dell'Italia, and P. A. Lucchesi Differential MAP kinase activation and Na+/H+ exchanger phosphorylation by H2O2 in rat cardiac myocytes Am J Physiol Cell Physiol, November 1, 2001; 281(5): C1542 - C1550. [Abstract] [Full Text] [PDF]

				R. A. Houliston, J. D. Pearson, and C. P. D. Wheeler-Jones Agonist-specific cross talk between ERKs and p38mapk regulates PGI2 synthesis in endothelium Am J Physiol Cell Physiol, October 1, 2001; 281(4): C1266 - C1276. [Abstract] [Full Text] [PDF]

				B. McLaughlin, S. Pal, M. P. Tran, A. A. Parsons, F. C. Barone, J. A. Erhardt, and E. Aizenman p38 Activation Is Required Upstream of Potassium Current Enhancement and Caspase Cleavage in Thiol Oxidant-Induced Neuronal Apoptosis J. Neurosci., May 15, 2001; 21(10): 3303 - 3311. [Abstract] [Full Text] [PDF]

				J. A. Aguirre-Ghiso, D. Liu, A. Mignatti, K. Kovalski, and L. Ossowski Urokinase Receptor and Fibronectin Regulate the ERKMAPK to p38MAPK Activity Ratios That Determine Carcinoma Cell Proliferation or Dormancy In Vivo Mol. Biol. Cell, April 1, 2001; 12(4): 863 - 879. [Abstract] [Full Text]

				R. M. Touyz, G. He, M. El Mabrouk, and E. L. Schiffrin p38 MAP Kinase Regulates Vascular Smooth Muscle Cell Collagen Synthesis by Angiotensin II in SHR But Not in WKY Hypertension, February 1, 2001; 37(2): 574 - 580. [Abstract] [Full Text] [PDF]

				R. M. Touyz and E. L. Schiffrin Signal Transduction Mechanisms Mediating the Physiological and Pathophysiological Actions of Angiotensin II in Vascular Smooth Muscle Cells Pharmacol. Rev., December 1, 2000; 52(4): 639 - 672. [Abstract] [Full Text] [PDF]

				T. HANNKEN, R. SCHROEDER, G. ZAHNER, R. A. K. STAHL, and G. WOLF Reactive Oxygen Species Stimulate p44/42 Mitogen-Activated Protein Kinase and Induce p27Kip1: Role in Angiotensin II-Mediated Hypertrophy of Proximal Tubular Cells J. Am. Soc. Nephrol., August 1, 2000; 11(8): 1387 - 1397. [Abstract] [Full Text]

				S. Kim and H. Iwao Molecular and Cellular Mechanisms of Angiotensin II-Mediated Cardiovascular and Renal Diseases Pharmacol. Rev., March 1, 2000; 52(1): 11 - 34. [Abstract] [Full Text] [PDF]

				A. K. Snabaitis, H. Yokoyama, and M. Avkiran Roles of Mitogen-Activated Protein Kinases and Protein Kinase C in {alpha}1A-Adrenoceptor-Mediated Stimulation of the Sarcolemmal Na+-H+ Exchanger Circ. Res., February 4, 2000; 86(2): 214 - 220. [Abstract] [Full Text] [PDF]

				J. Tan, T. Town, M. Saxe, D. Paris, Y. Wu, and M. Mullan Ligation of Microglial CD40 Results in p44/42 Mitogen-Activated Protein Kinase-Dependent TNF-{alpha} Production That Is Opposed by TGF-{beta}1 and IL-10 J. Immunol., December 15, 1999; 163(12): 6614 - 6621. [Abstract] [Full Text] [PDF]

				R. S. Haworth, J. Sinnett-Smith, E. Rozengurt, and M. Avkiran Protein kinase D inhibits plasma membrane Na+/H+ exchanger activity Am J Physiol Cell Physiol, December 1, 1999; 277(6): C1202 - C1209. [Abstract] [Full Text] [PDF]

				S. Gunasegaram, R. S. Haworth, D. J. Hearse, and M. Avkiran Regulation of Sarcolemmal Na+/H+ Exchanger Activity by Angiotensin II in Adult Rat Ventricular Myocytes : Opposing Actions via AT1 Versus AT2 Receptors Circ. Res., November 12, 1999; 85(10): 919 - 930. [Abstract] [Full Text] [PDF]

				O. Traub, T. Ishida, M. Ishida, J. C. Tupper, and B. C. Berk Shear Stress-mediated Extracellular Signal-regulated Kinase Activation Is Regulated by Sodium in Endothelial Cells. POTENTIAL ROLE FOR A VOLTAGE-DEPENDENT SODIUM CHANNEL J. Biol. Chem., July 16, 1999; 274(29): 20144 - 20150. [Abstract] [Full Text] [PDF]

				E. Takahashi, J.-i. Abe, B. Gallis, R. Aebersold, D. J. Spring, E. G. Krebs, and B. C. Berk p90RSK Is a Serum-stimulated Na+/H+ Exchanger Isoform-1 Kinase. REGULATORY PHOSPHORYLATION OF SERINE 703 OF Na+/H+ EXCHANGER ISOFORM-1 J. Biol. Chem., July 16, 1999; 274(29): 20206 - 20214. [Abstract] [Full Text] [PDF]

				Q. Jing, S.-M. Xin, Z.-J. Cheng, W.-B. Zhang, R. Zhang, Y.-W. Qin, and G. Pei Activation of p38 Mitogen-Activated Protein Kinase by Oxidized LDL in Vascular Smooth Muscle Cells : Mediation via Pertussis Toxin–Sensitive G Proteins and Association With Oxidized LDL-Induced Cytotoxicity Circ. Res., April 16, 1999; 84(7): 831 - 839. [Abstract] [Full Text] [PDF]

				S. Eguchi, P. J. Dempsey, G. D. Frank, E. D. Motley, and T. Inagami Activation of MAPKs by Angiotensin II in Vascular Smooth Muscle Cells. METALLOPROTEASE-DEPENDENT EGF RECEPTOR ACTIVATION IS REQUIRED FOR ACTIVATION OF ERK AND p38 MAPK BUT NOT FOR JNK J. Biol. Chem., March 9, 2001; 276(11): 7957 - 7962. [Abstract] [Full Text] [PDF]

				Y. V. Mukhin, T. Vlasova, A. A. Jaffa, G. Collinsworth, J. L. Bell, B. G. Tholanikunnel, T. Pettus, W. Fitzgibbon, D. W. Ploth, J. R. Raymond, et al. Bradykinin B2 Receptors Activate Na+/H+ Exchange in mIMCD-3 Cells via Janus Kinase 2 and Ca2+/Calmodulin J. Biol. Chem., May 11, 2001; 276(20): 17339 - 17346. [Abstract] [Full Text] [PDF]

				S. Lehoux, J.-i. Abe, J. A. Florian, and B. C. Berk 14-3-3 Binding to Na+/H+ Exchanger Isoform-1 Is Associated with Serum-dependent Activation of Na+/H+ Exchange J. Biol. Chem., May 4, 2001; 276(19): 15794 - 15800. [Abstract] [Full Text] [PDF]

				A. N. Moor, X. T. Gan, M. Karmazyn, and L. Fliegel Activation of Na+/H+ Exchanger-directed Protein Kinases in the Ischemic and Ischemic-reperfused Rat Myocardium J. Biol. Chem., May 4, 2001; 276(19): 16113 - 16122. [Abstract] [Full Text] [PDF]

				U. Schmitz, K. Thommes, I. Beier, W. Wagner, A. Sachinidis, R. Dusing, and H. Vetter Angiotensin II-induced Stimulation of p21-activated Kinase and c-Jun NH2-terminal Kinase Is Mediated by Rac1 and Nck J. Biol. Chem., June 15, 2001; 276(25): 22003 - 22010. [Abstract] [Full Text] [PDF]