Roles of NADPH oxidase 2 and 4 in endothelial cell survival and death under serum depletion
ABSTRACT
Oxidative stress and redox-signal pathways are known to be involved in endothelial apoptosis induced by serum depletion. However, the associated mechanism is not well understood and thus, was investigated in the present study focusing on NADPH oxidases (NOX). Serum removal from the culture medium led to an increase in reactive oxygen species (ROS) production and apoptotic death of human umbilical vein endothelial cells. Serum depletion also increased the gene expression of the NOX2 and NOX4 subunits. The selective suppression of NOX4 expression by small interfering RNA (siRNA) attenuated ROS production and cell death due to serum-depletion whereas siRNA for NOX2 increased cell death. Expression of exogenous NOX2 or NOX4 subunit alone had no significant effects on ROS production or cell death. Coexpression of the subunits of the NOX4 complex (NOX4 and p22phox) or the NOX2 complex (NOX2, p22phox, p47phox and p67phox) increased ROS production and cell death under serum-depleted conditions. This study suggests that endothelial cell survival and death are differentially regulated by expression levels of the subunits of NOX2 and NOX4 complexes.
Oxidative stress and redox-signal pathways are known to be involved in endothelial apoptosis induced by serum depletion. However, the associated mechanism is not well understood and thus, was investigated in the present study focusing on NADPH oxidases (NOX). Serum removal from the culture medium led to an increase in reactive oxygen species (ROS) production and apoptotic death of human umbilical vein endothelial cells. Serum depletion also increased the gene expression of the NOX2 and NOX4 subunits. The selective suppression of NOX4 expression by small interfering RNA (siRNA) attenuated ROS production and cell death due to serum-depletion whereas siRNA for NOX2 increased cell death. Expression of exogenous NOX2 or NOX4 subunit alone had no significant effects on ROS production or cell death. Coexpression of the subunits of the NOX4 complex (NOX4 and p22phox) or the NOX2 complex (NOX2, p22phox, p47phox and p67phox) increased ROS production and cell death under serum-depleted conditions. This study suggests that endothelial cell survival and death are differentially regulated by expression levels of the subunits of NOX2 and NOX4 complexes.
Cite this paper
Jeon, H. and Boo, Y. (2014) Roles of NADPH oxidase 2 and 4 in endothelial cell survival and death under serum depletion. Advances in Biological Chemistry, 4, 10-19. doi: 10.4236/abc.2014.41003.
Jeon, H. and Boo, Y. (2014) Roles of NADPH oxidase 2 and 4 in endothelial cell survival and death under serum depletion. Advances in Biological Chemistry, 4, 10-19. doi: 10.4236/abc.2014.41003.
References
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http://dx.doi.org/10.1016/j.freeradbiomed.2007.06.001 [13] Bedard, K. and Krause, K.H. (2007) The NOX family of ROS-generating NADPH oxidases: Physiology and pathophysiology. Physiological Reviews, 87, 245-313.
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http://dx.doi.org/10.1152/ajpcell.00381.2008 [19] Peshavariya, H., Dusting, G.J., Jiang, F., Halmos, L.R., Sobey, C.G., Drummond, G.R. and Selemidis, S. (2009) NADPH oxidase isoform selective regulation of endothelial cell proliferation and survival. Naunyn-Schmiedeberg’s Archives of Pharmacology, 380, 193-204.
http://dx.doi.org/10.1007/s00210-009-0413-0 [20] Deshpande, S.S., Angkeow, P., Huang, J., Ozaki, M. and Irani, K. (2000) Rac1 inhibits TNF-alpha-induced endothelial cell apoptosis: Dual regulation by reactive oxygen species. The FASEB Journal, 14, 1705-1714.
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http://dx.doi.org/10.1038/sj.onc.1209406 [22] Jeon, H. and Boo, Y.C. (2013) Laminar shear stress enhances endothelial cell survival through a NADPH oxidase 2-dependent mechanism. Biochemical and Biophysical Research Communications, 430, 460-465.
http://dx.doi.org/10.1016/j.bbrc.2012.12.016 [23] Petry, A., Djordjevic, T., Weitnauer, M., Kietzmann, T., Hess, J. and Gorlach, A. (2006) NOX2 and NOX4 mediate proliferative response in endothelial cells. Antioxidants & Redox Signaling, 8, 1473-1484.
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http://dx.doi.org/10.1016/j.abb.2003.07.006 [25] Buettner, G.R., Ng, C.F., Wang, M., Rodgers, V.G. and Schafer, F.Q. (2006) A new paradigm: Manganese superoxide dismutase influences the production of H2O2 in cells and thereby their biological state. Free Radical Biology & Medicine, 41, 1338-1350.
http://dx.doi.org/10.1016/j.freeradbiomed.2006.07.015 [26] Clement, M.V. and Pervaiz, S. (2001) Intracellular superoxide and hydrogen peroxide concentrations: A critical balance that determines survival or death. Redox Report, 6, 211-214.
http://dx.doi.org/10.1016/j.freeradbiomed.2006.07.015
[1] Gimbrone Jr., M.A. and Garcia-Cardena G. (2013) Vascular endothelium, hemodynamics, and the pathobiology of atherosclerosis. Cardiovascular Pathology, 22, 9-15.
http://dx.doi.org/10.1016/j.carpath.2012.06.006 [2] Furchgott, R.F. and Zawadzki, J.V. (1980) The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature, 288, 373-376.
http://dx.doi.org/10.1038/288373a0 [3] Libby, P. (2002) Inflammation in atherosclerosis. Nature, 420, 868-874.
http://dx.doi.org/10.1038/nature01323 [4] Giannotti, G. and Landmesser U. (2007) Endothelial dysfunction as an early sign of atherosclerosis. Herz Kardiovaskuläre Erkrankungen, 32, 568-572.
http://dx.doi.org/10.1007/s00059-007-3073-1 [5] Cai, H. and Harrison, D.G. (2000) Endothelial dysfunction in cardiovascular diseases: the role of oxidant stress. Circulation Research, 87, 840-844.
http://dx.doi.org/10.1161/01.RES.87.10.840 [6] Hadi, H.A., Carr, C.S. and Al Suwaidi, J. (2005) Endothelial dysfunction: Cardiovascular risk factors, therapy, and outcome. Vascular Health and Risk Management, 1, 183-198. [7] Dimmeler, S., Hermann C. and Zeiher, A.M. (1998) Apoptosis of endothelial cells. Contribution to the patho-physiology of atherosclerosis? European Cytokine Network, 9, 697-698. [8] Duval, H., Harris, M., Li, J., Johnson, N. and Print, C. (2003) New insights into the function and regulation of endothelial cell apoptosis. Angiogenesis, 6, 171-183.
http://dx.doi.org/10.1023/B:AGEN.0000021390.09275.bc [9] Tricot, O., Mallat, Z., Heymes, C., Belmin, J., Leseche, G. and Tedgui A. (2000) Relation between endothelial cell apoptosis and blood flow direction in human atherosclerotic plaques. Circulation, 101, 2450-2453.
http://dx.doi.org/10.1161/01.CIR.101.21.2450 [10] Kono, Y., Sawada, S., Kawahara, T., Tsuda, Y., Higaki, T., Yamasaki, S., Imamura, H., Tada, Y., Sato, T., Hiranuma, O., Akamatsu, N., Komatsu, S., Tamagaki, T., Nakagawa, K., Tsuji, H. and Nakagawa, M. (2002) Bradykinin inhibits serum-depletion-induced apoptosis of human vascular endothelial cells by inducing nitric oxide via calcium ion kinetics. Journal of Cardiovascular Pharmacology, 39, 251-261.
http://dx.doi.org/10.1097/00005344-200202000-00012 [11] Aoki, M., Nata, T., Morishita, R., Matsushita, H., Nakagami, H., Yamamoto, K., Yamazaki, K., Nakabayashi, M., Ogihara, T. and Kaneda, Y. (2001) Endothelial apoptosis induced by oxidative stress through activation of NF-kappaB: Antiapoptotic effect of antioxidant agents on endothelial cells. Hypertension, 38, 48-55.
http://dx.doi.org/10.1161/01.HYP.38.1.48 [12] Li, J.M., Fan, L.M., George, V.T. and Brooks, G. (2007) NOX2 regulates endothelial cell cycle arrest and apoptosis via p21cip1 and p53. Free Radical Biology & Medicine, 43, 976-986.
http://dx.doi.org/10.1016/j.freeradbiomed.2007.06.001 [13] Bedard, K. and Krause, K.H. (2007) The NOX family of ROS-generating NADPH oxidases: Physiology and pathophysiology. Physiological Reviews, 87, 245-313.
http://dx.doi.org/10.1152/physrev.00044.2005 [14] Lassegue, B. and Griendling, K.K. (2010) NADPH oxidases: Functions and pathologies in the vasculature. Arteriosclerosis, Thrombosis, and Vascular Biology, 30, 653-661.
http://dx.doi.org/10.1161/ATVBAHA.108.181610 [15] Martyn, K.D., Frederick, L.M., von Loehneysen, K., Dinauer, M.C. and Knaus, U.G. (2006) Functional analysis of Nox4 reveals unique characteristics compared to other NADPH oxidases. Cell Signaling, 18, 69-82.
http://dx.doi.org/10.1016/j.cellsig.2005.03.023 [16] Dikalov, S.I., Dikalova, A.E., Bikineyeva, A.T., Schmidt, H.H., Harrison, D.G. and Griendling, K.K. (2008) Distinct roles of Nox1 and NOX4 in basal and angiotensin II-stimulated superoxide and hydrogen peroxide production. Free Radical Biology & Medicine, 45, 1340-1351.
http://dx.doi.org/10.1016/j.freeradbiomed.2008.08.013 [17] Irani, K. (2000) Oxidant signaling in vascular cell growth, death, and survival: A review of the roles of reactive oxygen species in smooth muscle and endothelial cell mitogenic and apoptotic signaling. Circulation Research, 87, 179-183.
http://dx.doi.org/10.1161/01.RES.87.3.179 [18] Basuroy, S., Bhattacharya, S., Leffler, C.W. and Parfenova, H. (2009) NOX4 NADPH oxidase mediates oxidative stress and apoptosis caused by TNF-alpha in cerebral vascular endothelial cells. American Journal of Physiology-Cell Physiology, 296, C422-C432.
http://dx.doi.org/10.1152/ajpcell.00381.2008 [19] Peshavariya, H., Dusting, G.J., Jiang, F., Halmos, L.R., Sobey, C.G., Drummond, G.R. and Selemidis, S. (2009) NADPH oxidase isoform selective regulation of endothelial cell proliferation and survival. Naunyn-Schmiedeberg’s Archives of Pharmacology, 380, 193-204.
http://dx.doi.org/10.1007/s00210-009-0413-0 [20] Deshpande, S.S., Angkeow, P., Huang, J., Ozaki, M. and Irani, K. (2000) Rac1 inhibits TNF-alpha-induced endothelial cell apoptosis: Dual regulation by reactive oxygen species. The FASEB Journal, 14, 1705-1714.
http://dx.doi.org/10.1096/fj.99-0910com [21] Mochizuki, T., Furuta S., Mitsushita J., Shang, W.H., Ito, M., Yokoo, Y., Yamaura, M., Ishizone, S., Nakayama, J., Konagai, A., Hirose, K., Kiyosawa, K. and Kamata T. (2006) Inhibition of NADPH oxidase 4 activates apoptosis via the AKT/apoptosis signal-regulating kinase 1 pathway in pancreatic cancer PANC-1 cells. Oncogene, 25, 3699-3707.
http://dx.doi.org/10.1038/sj.onc.1209406 [22] Jeon, H. and Boo, Y.C. (2013) Laminar shear stress enhances endothelial cell survival through a NADPH oxidase 2-dependent mechanism. Biochemical and Biophysical Research Communications, 430, 460-465.
http://dx.doi.org/10.1016/j.bbrc.2012.12.016 [23] Petry, A., Djordjevic, T., Weitnauer, M., Kietzmann, T., Hess, J. and Gorlach, A. (2006) NOX2 and NOX4 mediate proliferative response in endothelial cells. Antioxidants & Redox Signaling, 8, 1473-1484.
http://dx.doi.org/10.1089/ars.2006.8.1473 [24] Valgimigli, M., Merli, E., Malagutti, P., Soukhomovskaia, O., Cicchitelli, G., Macri, G. and Ferrari, R. (2003) Endothelial dysfunction in acute and chronic coronary syndromes: Evidence for a pathogenetic role of oxidative stress. Archives of Biochemistry and Biophysics, 420, 255-261.
http://dx.doi.org/10.1016/j.abb.2003.07.006 [25] Buettner, G.R., Ng, C.F., Wang, M., Rodgers, V.G. and Schafer, F.Q. (2006) A new paradigm: Manganese superoxide dismutase influences the production of H2O2 in cells and thereby their biological state. Free Radical Biology & Medicine, 41, 1338-1350.
http://dx.doi.org/10.1016/j.freeradbiomed.2006.07.015 [26] Clement, M.V. and Pervaiz, S. (2001) Intracellular superoxide and hydrogen peroxide concentrations: A critical balance that determines survival or death. Redox Report, 6, 211-214.
http://dx.doi.org/10.1016/j.freeradbiomed.2006.07.015