© 1995 American Heart Association, Inc.
Articles |
Fluid Shear Stress Stimulates Mitogen-Activated Protein Kinase in Endothelial Cells
From the Department of Physiology, Emory University, Atlanta, Ga (H.T.), and the Department of Medicine (Cardiology Division), University of Washington, Seattle.
Correspondence to Bradford C. Berk, MD, PhD, University of Washington, Cardiology Division, Box 357710, Seattle, WA 98195. E-mail beberk@u.washington.edu.
Abstract Local alterations in the hemodynamic environment regulate endothelial cell function, but the signal-transduction mechanisms involved in this process remain unclear. Because mitogen-activated protein (MAP) kinases have been shown to be activated by physical forces, we measured the phosphorylation and enzyme activity of MAP kinase to identify the signal events involved in the endothelial cell response to fluid shear stress. Flow at physiological shear stress (3.5 to 117 dynes/cm2) activated 42-kD and 44-kD MAP kinases present in cultured bovine aortic endothelial cells, with maximal effect at 12 dynes/cm2. Activation of a G protein was necessary, as demonstrated by complete inhibition by the nonhydrolyzable GDP analog GDP-ßS. Activation of protein kinase C (PKC) was required, as shown by inhibiting PKC with staurosporine or downregulating PKC with phorbol 12,13-dibutyrate. Both Ca2+-dependent and -independent PKC activity, measured by translocation and substrate phosphorylation, increased in response to flow. However, MAP kinase activation was not dependent on Ca2+ mobilization, since Ca2+ chelation had no inhibitory effect. On the basis of these findings, it is proposed that flow activates two signal-transduction pathways in endothelial cells. One pathway is Ca2+ dependent and involves activation of phospholipase C and increases in intracellular Ca2+. A new pathway, described in the present study, is Ca2+ independent and involves a G protein and increases in PKC and MAP kinase activity.
Key Words: endothelium • signal transduction • MAP kinase • protein kinase C • shear stress
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E. M. Redmond, P. A. Cahill, and J. V. Sitzmann Flow-Mediated Regulation of G-Protein Expression in Cocultured Vascular Smooth Muscle and Endothelial Cells Arterioscler Thromb Vasc Biol, January 1, 1998; 18(1): 75 - 83. [Abstract] [Full Text] [PDF]
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S. Chien, S. Li, and J. Y-J. Shyy Effects of Mechanical Forces on Signal Transduction and Gene Expression in Endothelial Cells Hypertension, January 1, 1998; 31(1): 162 - 169. [Abstract] [Full Text] [PDF]
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K. G. Birukov, S. Lehoux, A. A. Birukova, R. Merval, V. A. Tkachuk, and A. Tedgui Increased Pressure Induces Sustained Protein Kinase C–Independent Herbimycin A–Sensitive Activation of Extracellular Signal–Related Kinase 1/2 in the Rabbit Aorta in Organ Culture Circ. Res., December 19, 1997; 81(6): 895 - 903. [Abstract] [Full Text]
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J. M. Pyles, K. L. March, M. Franklin, K. Mehdi, R. L. Wilensky, and L. P. Adam Activation of MAP Kinase In Vivo Follows Balloon Overstretch Injury of Porcine Coronary and Carotid Arteries Circ. Res., December 19, 1997; 81(6): 904 - 910. [Abstract] [Full Text]
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O. Traub, B. P. Monia, N. M. Dean, and B. C. Berk PKC-epsilon Is Required for Mechano-sensitive Activation of ERK1/2 in Endothelial Cells J. Biol. Chem., December 12, 1997; 272(50): 31251 - 31257. [Abstract] [Full Text] [PDF]
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J.J. Chiu, B.S. Wung, J. Y.J. Shyy, H.J. Hsieh, and D.L. Wang Reactive Oxygen Species Are Involved in Shear Stress-Induced Intercellular Adhesion Molecule-1 Expression in Endothelial Cells Arterioscler Thromb Vasc Biol, December 1, 1997; 17(12): 3570 - 3577. [Abstract] [Full Text]
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M. T. Franklin, C. L.-A. Wang, and L. P. Adam Stretch-dependent activation and desensitization of mitogen-activated protein kinase in carotid arteries Am J Physiol Cell Physiol, December 1, 1997; 273(6): C1819 - C1827. [Abstract] [Full Text] [PDF]
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S. Li, M. Kim, Y.-L. Hu, S. Jalali, D. D. Schlaepfer, T. Hunter, S. Chien, and J. Y-J. Shyy Fluid Shear Stress Activation of Focal Adhesion Kinase. LINKING TO MITOGEN-ACTIVATED PROTEIN KINASES J. Biol. Chem., November 28, 1997; 272(48): 30455 - 30462. [Abstract] [Full Text] [PDF]
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T. Caulin-Glaser, G. Garcia-Cardena, P. Sarrel, W. C. Sessa, and J. R. Bender 17ß-Estradiol Regulation of Human Endothelial Cell Basal Nitric Oxide Release, Independent of Cytosolic Ca2+ Mobilization Circ. Res., November 19, 1997; 81(5): 885 - 892. [Abstract] [Full Text]
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Y. Hu, L. Cheng, B.-W. Hochleitner, and Q. Xu Activation of Mitogen-Activated Protein Kinases (ERK/JNK) and AP-1 Transcription Factor in Rat Carotid Arteries After Balloon Injury Arterioscler Thromb Vasc Biol, November 1, 1997; 17(11): 2808 - 2816. [Abstract] [Full Text]
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L. M. Khachigian and T. Collins Inducible Expression of Egr-1–Dependent Genes : A Paradigm of Transcriptional Activation in Vascular Endothelium Circ. Res., October 19, 1997; 81(4): 457 - 461. [Full Text]
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J. K. Miyashiro, V. Poppa, and B. C. Berk Flow-Induced Vascular Remodeling in the Rat Carotid Artery Diminishes With Age Circ. Res., September 19, 1997; 81(3): 311 - 319. [Abstract] [Full Text]
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E. O. Harrington, J. Loffler, P. R. Nelson, K. C. Kent, M. Simons, and J. A. Ware Enhancement of Migration by Protein Kinase Calpha and Inhibition of Proliferation and Cell Cycle Progression by Protein Kinase Cdelta in Capillary Endothelial Cells J. Biol. Chem., March 14, 1997; 272(11): 7390 - 7397. [Abstract] [Full Text] [PDF]
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J. M. Muller, W. M. Chilian, and M. J. Davis Integrin Signaling Transduces Shear Stress–Dependent Vasodilation of Coronary Arterioles Circ. Res., March 1, 1997; 80(3): 320 - 326. [Abstract] [Full Text]
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H. Jo, K. Sipos, Y.-M. Go, R. Law, J. Rong, and J. M. McDonald Differential Effect of Shear Stress on Extracellular Signal-regulated Kinase and N-terminal Jun Kinase in Endothelial Cells. Gi2- AND Gbeta /gamma -DEPENDENT SIGNALING PATHWAYS J. Biol. Chem., January 10, 1997; 272(2): 1395 - 1401. [Abstract] [Full Text] [PDF]
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M. Kusuhara, A. Chait, A. Cader, and B. C. Berk Oxidized LDL Stimulates Mitogen-Activated Protein Kinases in Smooth Muscle Cells and Macrophages Arterioscler Thromb Vasc Biol, January 1, 1997; 17(1): 141 - 148. [Abstract] [Full Text]
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M. A. Corson, N. L. James, S. E. Latta, R. M. Nerem, B. C. Berk, and D. G. Harrison Phosphorylation of Endothelial Nitric Oxide Synthase in Response to Fluid Shear Stress Circ. Res., November 1, 1996; 79(5): 984 - 991. [Abstract] [Full Text]
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S. R.P. Gudi, C. B. Clark, and J. A. Frangos Fluid Flow Rapidly Activates G Proteins in Human Endothelial Cells: Involvement of G Proteins in Mechanochemical Signal Transduction Circ. Res., October 1, 1996; 79(4): 834 - 839. [Abstract] [Full Text]
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T. Ishida, T. E. Peterson, N. L. Kovach, and B. C. Berk MAP Kinase Activation by Flow in Endothelial Cells: Role of ß1 Integrins and Tyrosine Kinases Circ. Res., August 1, 1996; 79(2): 310 - 316. [Abstract] [Full Text]
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J.-i. Abe, M. Kusuhara, R. J. Ulevitch, B. C. Berk, and J.-D. Lee Big Mitogen-activated Protein Kinase 1(BMK1) Is a Redox-sensitive Kinase J. Biol. Chem., July 12, 1996; 271(28): 16586 - 16590. [Abstract] [Full Text] [PDF]
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K. Ayajiki, M. Kindermann, M. Hecker, I. Fleming, and R. Busse Intracellular pH and Tyrosine Phosphorylation but Not Calcium Determine Shear Stress–Induced Nitric Oxide Production in Native Endothelial Cells Circ. Res., May 1, 1996; 78(5): 750 - 758. [Abstract] [Full Text]
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P. F. Davies NO Flow Helps Clear Murky Waters? Circ. Res., May 1, 1996; 78(5): 945 - 946. [Full Text]
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