Int J Angiol 2017; 26(02): 109-115
DOI: 10.1055/s-0037-1599057
Original Article
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Shear Stress Induces Change in Extracellular Signal-Regulated Kinase 5 Levels with Sustained Activation under Disturbed and Continuous Laminar Flow

S. Y. Shalaby
1   Section of Vascular Surgery, Yale University, New Haven, Connecticut
,
G. Chitragari
1   Section of Vascular Surgery, Yale University, New Haven, Connecticut
,
B. J. Sumpio
1   Section of Vascular Surgery, Yale University, New Haven, Connecticut
,
B. E. Sumpio
1   Section of Vascular Surgery, Yale University, New Haven, Connecticut
› Author Affiliations
Further Information

Publication History

Publication Date:
25 February 2017 (online)

Abstract

Extracellular signal-regulated kinase 5 (ERK5) has been reported to regulate endothelial integrity and protect from vascular dysfunction under laminar flow. Previously reported research indicates that under laminar flow ERK5 is activated with production of atheroprotective molecules. However, the characterization of ERK5 activation and levels under different flow patterns has not been investigated.

Confluent HUVECs were serum-starved then seeded on glass slides. HUVECs incubated in 1% FBS were exposed to continuous laminar flow (CLF), to-and-fro flow (TFF), or pulsatile forward flow (PFF) in a parallel plate flow chamber. At the end of experimentation, cell lysates were immunoblotted with antibodies to phospho-ERK5 and total ERK5. ERK5 activation was assessed by the levels of phosphorylated ERK5. The densitometric mean ± SEM is calculated and analyzed by ANOVA. p < 0.05 is considered significant.

Levels of ERK5 decreased with all flow conditions with the largest decrease in TFF flow condition. TFF and CLF exhibited sustained ERK5 phosphorylation in HUVECs stimulated for up to 4 hours. PFF had transient phosphorylation of ERK5 at 2 hours, which then became undetectable at 4 hours of exposure to flow. Also, TFF and CLF both showed decreased levels at 4 hours, suggesting a decrease in activation for these flow conditions.

Exposure of HUVEC to different types of shear stress results in varying patterns of activation of ERK5. Activation of ERK5 with TFF suggests a role in the pathogenesis of atherosclerosis and vascular remodeling under disturbed flow conditions.

 
  • References

  • 1 VanderLaan PA, Reardon CA, Getz GS. Site specificity of atherosclerosis: site-selective responses to atherosclerotic modulators. Arterioscler Thromb Vasc Biol 2004; 24 (1) 12-22
  • 2 Frangos SG, Gahtan V, Sumpio B. Localization of atherosclerosis: role of hemodynamics. Arch Surg 1999; 134 (10) 1142-1149
  • 3 Hahn C, Schwartz MA. Mechanotransduction in vascular physiology and atherogenesis. Nat Rev Mol Cell Biol 2009; 10 (1) 53-62
  • 4 Berk BC. Atheroprotective signaling mechanisms activated by steady laminar flow in endothelial cells. Circulation 2008; 117 (8) 1082-1089
  • 5 Sumpio BE, Yun S, Cordova AC , et al. MAPKs (ERK1/2, p38) and AKT can be phosphorylated by shear stress independently of platelet endothelial cell adhesion molecule-1 (CD31) in vascular endothelial cells. J Biol Chem 2005; 280 (12) 11185-11191
  • 6 Berk BC, Abe JI, Min W, Surapisitchat J, Yan C. Endothelial atheroprotective and anti-inflammatory mechanisms. Ann N Y Acad Sci 2001; 947: 93-109 , discussion 109–111
  • 7 Chang L, Karin M. Mammalian MAP kinase signalling cascades. Nature 2001; 410 (6824): 37-40
  • 8 Azuma N, Duzgun SA, Ikeda M , et al. Endothelial cell response to different mechanical forces. J Vasc Surg 2000; 32 (4) 789-794
  • 9 Yan C, Takahashi M, Okuda M, Lee JD, Berk BC. Fluid shear stress stimulates big mitogen-activated protein kinase 1 (BMK1) activity in endothelial cells. Dependence on tyrosine kinases and intracellular calcium. J Biol Chem 1999; 274 (1) 143-150
  • 10 Nigro P, Abe J, Woo CH , et al. PKCzeta decreases eNOS protein stability via inhibitory phosphorylation of ERK5. Blood 2010; 116 (11) 1971-1979
  • 11 Abe R, Yamashita N, Rochier A , et al. Pulsatile to-fro flow induces greater and sustained expression of tissue factor RNA in HUVEC than unidirectional laminar flow. Am J Physiol Heart Circ Physiol 2011; 300 (4) H1345-H1351
  • 12 Azuma N, Akasaka N, Kito H , et al. Role of p38 MAP kinase in endothelial cell alignment induced by fluid shear stress. Am J Physiol Heart Circ Physiol 2001; 280 (1) H189-H197
  • 13 Suzaki Y, Yoshizumi M, Kagami S , et al. Hydrogen peroxide stimulates c-Src-mediated big mitogen-activated protein kinase 1 (BMK1) and the MEF2C signaling pathway in PC12 cells: potential role in cell survival following oxidative insults. J Biol Chem 2002; 277 (11) 9614-9621
  • 14 Ohnesorge N, Viemann D, Schmidt N , et al. Erk5 activation elicits a vasoprotective endothelial phenotype via induction of Kruppel-like factor 4 (KLF4). J Biol Chem 2010; 285 (34) 26199-26210
  • 15 Hamik A, Lin Z, Kumar A , et al. Kruppel-like factor 4 regulates endothelial inflammation. J Biol Chem 2007; 282 (18) 13769-13779
  • 16 McCormick SM, Eskin SG, McIntire LV , et al. DNA microarray reveals changes in gene expression of shear stressed human umbilical vein endothelial cells. Proc Natl Acad Sci U S A 2001; 98 (16) 8955-8960
  • 17 Komaravolu RK, Adam C, Moonen JR, Harmsen MC, Goebeler M, Schmidt M. Erk5 inhibits endothelial migration via KLF2-dependent down-regulation of PAK1. Cardiovasc Res 2015; 105 (1) 86-95
  • 18 Parmar KM, Larman HB, Dai G , et al. Integration of flow-dependent endothelial phenotypes by Kruppel-like factor 2. J Clin Invest 2006; 116 (1) 49-58
  • 19 Gavine PR, Wang M, Yu D , et al. Identification and validation of dysregulated MAPK7 (ERK5) as a novel oncogenic target in squamous cell lung and esophageal carcinoma. BMC Cancer 2015; 15: 454
  • 20 Yoshizumi M, Kyotani Y, Zhao J , et al. Role of big mitogen-activated protein kinase 1 (BMK1)/extracellular signal-regulated kinase 5 (ERK5) in the pathogenesis and progression of atherosclerosis. J Pharmacol Sci 2012; 120 (4) 259-263
  • 21 Lochhead PA, Gilley R, Cook SJ. ERK5 and its role in tumour development. Biochem Soc Trans 2012; 40 (1) 251-256
  • 22 Wang X, Merritt AJ, Seyfried J , et al. Targeted deletion of mek5 causes early embryonic death and defects in the extracellular signal-regulated kinase 5/myocyte enhancer factor 2 cell survival pathway. Mol Cell Biol 2005; 25 (1) 336-345
  • 23 Pi X, Yan C, Berk BC. Big mitogen-activated protein kinase (BMK1)/ERK5 protects endothelial cells from apoptosis. Circ Res 2004; 94 (3) 362-369
  • 24 Sohn SJ, Sarvis BK, Cado D, Winoto A. ERK5 MAPK regulates embryonic angiogenesis and acts as a hypoxia-sensitive repressor of vascular endothelial growth factor expression. J Biol Chem 2002; 277 (45) 43344-43351
  • 25 Hayashi M, Kim SW, Imanaka-Yoshida K , et al. Targeted deletion of BMK1/ERK5 in adult mice perturbs vascular integrity and leads to endothelial failure. J Clin Invest 2004; 113 (8) 1138-1148
  • 26 Li L, Tatake RJ, Natarajan K , et al. Fluid shear stress inhibits TNF-mediated JNK activation via MEK5-BMK1 in endothelial cells. Biochem Biophys Res Commun 2008; 370 (1) 159-163
  • 27 Heo KS, Chang E, Le NT , et al. De-SUMOylation enzyme of sentrin/SUMO-specific protease 2 regulates disturbed flow-induced SUMOylation of ERK5 and p53 that leads to endothelial dysfunction and atherosclerosis. Circ Res 2013; 112 (6) 911-923
  • 28 Conway DE, Williams MR, Eskin SG, McIntire LV. Endothelial cell responses to atheroprone flow are driven by two separate flow components: low time-average shear stress and fluid flow reversal. Am J Physiol Heart Circ Physiol 2010; 298 (2) H367-H374