Characterization of Extracellular Signal-Regulated Kinase 5 Levels in Human Umbilical Vein Endothelial Cells Exposed to Disturbed and Uniform Flow

Abstract

Extracellular signal-regulated kinase 5 (ERK5) has been reported to regulate endothelial cell integrity and protect from vascular dysfunction under continuous laminar flow. However, the effect of flow on ERK5 levels has not been determined. Confluent human umbilical vein endothelial cells (HUVECs) were seeded on fibronectin coated glass slides and serum starved for 2 hours with 1% fetal bovine serum (FBS). HUVECs were then exposed to to and fro flow (TFF), pulsatile forward flow (PFF), or continuous laminar flow (CLF) in a parallel plate flow chamber for up to 2 hours. At the end of experiment, cell lysates were prepared and immunoblotted with antibodies to total ERK5. Both CLF and TFF exhibited a decrease in ERK5 after levels after 2-hour exposure. However, the level of ERK5 for PFF remained the same. Disturbed, but not uniform pulsatile, flow decreases ERK5 levels in HUVECs.

Keywords

pulsatile uniform flow - atherosclerosis - HUVECs - ERK5 - disturbed flow - laminar flow - uniform flow

• 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 Gimbrone Jr MA, Topper JN, Nagel T, Anderson KR, Garcia-Cardeña G. Endothelial dysfunction, hemodynamic forces, and atherogenesis. Ann N Y Acad Sci 2000; 902: 230-239 , discussion 239–240
• 3 Berk BC. Atheroprotective signaling mechanisms activated by steady laminar flow in endothelial cells. Circulation 2008; 117 (8) 1082-1089
• 4 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
• 5 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
• 6 Chang L, Karin M. Mammalian MAP kinase signalling cascades. Nature 2001; 410 (6824) 37-40
• 7 Azuma N, Duzgun SA, Ikeda M , et al. Endothelial cell response to different mechanical forces. J Vasc Surg 2000; 32 (4) 789-794
• 8 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
• 9 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
• 10 Zarins CK, Giddens DP, Bharadvaj BK, Sottiurai VS, Mabon RF, Glagov S. Carotid bifurcation atherosclerosis. Quantitative correlation of plaque localization with flow velocity profiles and wall shear stress. Circ Res 1983; 53 (4) 502-514
• 11 Gimbrone Jr MA, García-Cardeña G. Vascular endothelium, hemodynamics, and the pathobiology of atherosclerosis. Cardiovasc Pathol 2013; 22 (1) 9-15
• 12 Nithianandarajah-Jones GN, Wilm B, Goldring CE, Müller J, Cross MJ. ERK5: structure, regulation and function. Cell Signal 2012; 24 (11) 2187-2196
• 13 Kasler HG, Victoria J, Duramad O, Winoto A. ERK5 is a novel type of mitogen-activated protein kinase containing a transcriptional activation domain. Mol Cell Biol 2000; 20 (22) 8382-8389
• 14 Kondoh K, Terasawa K, Morimoto H, Nishida E. Regulation of nuclear translocation of extracellular signal-regulated kinase 5 by active nuclear import and export mechanisms. Mol Cell Biol 2006; 26 (5) 1679-1690
• 15 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
• 16 Abe R, Yamashita N, Rochier A , et al. Varying effects of hemodynamic forces on tissue factor RNA expression in human endothelial cells. J Surg Res 2011; 170 (1) 150-156
• 17 Kim M, Kim S, Lim JH, Lee C, Choi HC, Woo CH. Laminar flow activation of ERK5 protein in vascular endothelium leads to atheroprotective effect via NF-E2-related factor 2 (Nrf2) activation. J Biol Chem 2012; 287 (48) 40722-40731
• 18 Spiering D, Schmolke M, Ohnesorge N , et al. MEK5/ERK5 signaling modulates endothelial cell migration and focal contact turnover. J Biol Chem 2009; 284 (37) 24972-24980