Shear stress effect on endothelial nitric oxide synthase in cultured human umbilical vein endothelial cells


Background: Low shear stress caused by disturbed or turbulent flow at arterial branch points is known to associate with atherosclerosis. However, shear stress at the venous valve location and its association with deep vein thrombosis are less understood due to the complex and poorly understood bi-directional flow in the valve pocket region. We investigated how venous endothelial cells respond to flow shear stress around the venous valve region using a novel in vitro system that mimics venous flow. Results: Human umbilical vein EAhy. 926 cells were cultured on a flexible silastic membrane that mimicked venous tissue. Confluent cells were exposed to sinusoidal uni-and bi-directional pulsatile shear stress (0.1 to 1 dyne/cm2) for up to 6 h. Western-blot analyses indicated that endothelial nitric oxide (eNOS) expression levels decreased regardless of all tested flow patterns, stress magnitude, and shearing time. In contrast, the expression levels of inhibitor of κB (kappa B) and α (alpha)-tubulin were unaffected by the shear stress. Conclusions: Our results indicate that shear stress causes a decrease specifically in eNOS expression, suggesting that it may play a significant role in regulating inflammation related protein expression in endothelial cells.

Share and Cite:

Gu, Q. , Smith, D. and Hoo, K. (2013) Shear stress effect on endothelial nitric oxide synthase in cultured human umbilical vein endothelial cells. Journal of Biomedical Science and Engineering, 6, 982-986. doi: 10.4236/jbise.2013.610122.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Aird, W. (2005) Spatial and temporal dynamics of the endothelium. Journal of Thrombosis and Haemostasis, 3, 1392-1406.
[2] Chatzizisis, Y.S., Coskun, A.U., Jonas, M., Edelman, E.R., Feldman, C.L. and Stone, P.H. (2007) Role of endothelial shear stress in the natural history of coronary atherosclerosis and vascular remodeling: Molecular, cellular, and vascular behavior. Journal of the American College of Cardiology, 49, 2379-2393.
[3] Aird, W.C. (2007) Phenotypic heterogeneity of the endothelium. Circulation research, 100, 174-190.
[4] Nagel, T., Resnick, N., Atkinson, W.J., Dewey Jr., C.F. and Gimbrone Jr., M.A. (1994) Shear stress selectively upregulates intercellular adhesion molecule-1 expression in cultured human vascular endothelial cells. Journal of Clinical Investigation, 94, 885.
[5] Barakat, A.I. and Lieu, D.K. (2003) Differential responsiveness of vascular endothelial cells to different types of fluid mechanical shear stress. Cell Biochemistry and Biophysics, 38, 323-343.
[6] Cirino, G., Fiorucci, S. and Sessa, W.C. (2003) Endothelial nitric oxide synthase: The Cinderella of inflammation? Trends in Pharmacological Sciences, 24, 91-95.
[7] Forstermann, U. and Li, H. (2011) Therapeutic effect of enhancing endothelial nitric oxide synthase (eNOS) expression and preventing eNOS uncoupling. British Journal of Pharmacology, 164, 213-223.
[8] Zhang, Y., Janssens, S.P., Wingler, K., Schmidt, H.H.H. W. and Moens, A.L. (2011) Modulating endothelial nitric oxide synthase: A new cardiovascular therapeutic strategy. American Journal of Physiology-Heart and Circulatory Physiology, 301, H634-H646.
[9] Cunningham, K.S. and Gotlieb, A.I. (2004) The role of shear stress in the pathogenesis of atherosclerosis. Laboratory Investigation, 85, 9-23.
[10] Quyyumi, A.A. (1998) Endothelial function in health and disease: New insights into the genesis of cardiovascular disease. The American Journal of Medicine, 105, 32S-39S.
[11] Nicolaides, A. (2000) Investigation of chronic venous insufficiency: A consensus statement. Circulation, 102, e126-e163.
[12] Hamer, J., Malone, P. and Silver, I. (1981) The PO2 in venous valve pockets: Its possible bearing on thrombogenesis. British Journal of Surgery, 68, 166-170.
[13] Karino, T. and Motomiya, M. (1984) Flow through a venous valve and its implication for thrombus formation. Thrombosis Research, 36, 245-257.
[14] Simpson, D., Majeski, M., Borg, T. and Terracio, L. (1999) Regulation of cardiac myocyte protein turnover and myofibrillar structure in vitro by specific directions of stretch. Circulation Research, 85, e59.
[15] Edgell, C.J.S., Haizlip, J.E. and Bagnell, C.R. (1990) Endothelium specific Weibel-Palade bodies in a continuous human cell line, EA. hy926. In Vitro Cellular & Developmental Biology-Plant, 26, 1167-1172.
[16] Edgell, C., McDonald, C.C. and Graham, J.B. (1983) Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proceedings of the National Academy of Sciences, 80, 3734.
[17] Lurie, F., Kistner, R.L., Eklof, B. and Kessler, D. (2003) Mechanism of venous valve closure and role of the valve in circulation: A new concept. Journal of Vascular Surgery, 38, 955-961.
[18] Guzik, T., Korbut, R. and Adamek-Guzik, T. (2003) Nitric oxide and superoxide in inflammation. Journal of Physiology and Pharmacology, 54, 469-487.
[19] Harrison, D.G., Sayegh, H., Ohara, Y., Inoue, N. and Venema, R.C. (1996) Regulation of expression of the endothelial cell nitric oxide synthase. Clinical and Experimental Pharmacology and Physiology, 23, 251-255.
[20] Li, Y., Haga, J. and Chien, S. (2005) Molecular basis of the effects of shear stress on vascular endothelial cells. Journal of Biomechanics, 38, 1949-1971.
[21] Papafaklis, M.I., Koskinas, K.C., Chatzizisis, Y.S., Stone, P.H. and Feldman, C.L. (2010) In vivo assessment of the natural history of coronary atherosclerosis: vascular remodeling and endothelial shear stress determine the complexity of atherosclerotic disease progression. Current Opinion in Cardiology, 25, 627.
[22] Malek, A.M., Alper, S.L. and Izumo, S. (1999) Hemodynamic shear stress and its role in atherosclerosis. The Journal of the American Medical Association, 282, 2035-2042.
[23] Casas, J.P., Bautista, L.E., Humphries, S.E. and Hingorani, A.D. (2004) Endothelial nitric oxide synthase genotype and ischemic heart disease. Circulation, 109, 1359-1365.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.