Published online before print January 31, 2002, doi: 10.1161/01.ATV.0000012262.76205.6A
© 2002 American Heart Association, Inc.
Vascular Biology |
Role of Mitochondrial Oxidant Generation in Endothelial Cell Responses to Hypoxia
From the Departments of Medicine and Surgery, The University of Chicago, Chicago, Ill.
Correspondence to Paul T. Schumacker, PhD, Department of Medicine, MC6026, 5841 S Maryland Ave, Chicago, IL 60637. E-mail pschumac{at}medicine.bsd.uchicago.edu
Endothelial cells increase their secretion of the cytokine interleukin-6 (IL-6) during hypoxia, which then acts in an autocrine fashion to increase the permeability of cell monolayers. These responses are attenuated by antioxidants, suggesting that reactive oxygen species (ROS) participate in signaling in hypoxic endothelium. We tested whether mitochondria are responsible for these ROS in human umbilical vein endothelial cells exposed to hypoxia. Oxidation of the probe 2', 7'-dichlorodihydrofluorescein to fluorescent dichlorofluorescein or the probe dihydroethidium was used to assess oxidant signaling, whereas permeability was assessed by using transendothelial electrical resistance. Hypoxia elicited increases in dichlorofluorescein and dihydroethidium fluorescence that were abrogated by the mitochondrial electron transport (ET) inhibitors rotenone (2 µmol/L) and diphenyleneiodonium (5 µmol/L). The same ET inhibitors also attenuated hypoxia-induced increases in nuclear factor-
B (NF-B) activation, although they did not abrogate NF-B activation in response to endotoxin (lipopolysaccharide). ET inhibition also abolished the hypoxia-induced increases in IL-6 mRNA expression, hypoxia-stimulated IL-6 secretion into the media, and the hypoxia-induced increases in transendothelial electrical resistance of human umbilical vein endothelial cell monolayers. By contrast, the above responses to hypoxia were not significantly affected by treatment with the NAD(P)H oxidase inhibitor apocynin (30 µmol/L), the xanthine oxidase inhibitor allopurinol (100 µmol/L), or the NO synthase inhibitor N-nitro-L-arginine (100 µmol/L). We conclude that ROS signals originating from the mitochondrial ET chain trigger the increase in NF-B activation, the transcriptional activation of IL-6, the secretion of IL-6 into the cell culture media, and the increases in endothelial permeability observed during hypoxia.
Key Words: reactive oxygen species • human umbilical vein endothelial cells • ischemia • signal transduction • microcirculation
This article has been cited by other articles:
 |
|
 |
|
  A. S. Godbole, X. Lu, X. Guo, and G. S. Kassab NADPH oxidase has a directional response to shear stress Am J Physiol Heart Circ Physiol, January 1, 2009; 296(1): H152 - H158. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. H. Ruder, T. J. Hartman, J. Blumberg, and M. B. Goldman Oxidative stress and antioxidants: exposure and impact on female fertility Hum. Reprod. Update, June 4, 2008; (2008) dmn011v1. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Baudry, E. Laemmel, and E. Vicaut In vivo reactive oxygen species production induced by ischemia in muscle arterioles of mice: involvement of xanthine oxidase and mitochondria Am J Physiol Heart Circ Physiol, February 1, 2008; 294(2): H821 - H828. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Ostergaard, E. Stankevicius, M. R. Andersen, Y. Eskildsen-Helmond, T. Ledet, M. J. Mulvany, and U. Simonsen Diminished NO release in chronic hypoxic human endothelial cells Am J Physiol Heart Circ Physiol, November 1, 2007; 293(5): H2894 - H2903. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. X. Zhang and D. D. Gutterman Mitochondrial reactive oxygen species-mediated signaling in endothelial cells Am J Physiol Heart Circ Physiol, May 1, 2007; 292(5): H2023 - H2031. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. M. Davidson and M. R. Duchen Endothelial Mitochondria: Contributing to Vascular Function and Disease Circ. Res., April 27, 2007; 100(8): 1128 - 1141. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Han, Y.-R. Chen, C. I. Jones III, G. Meenakshisundaram, J. L. Zweier, and B. R. Alevriadou Shear-induced reactive nitrogen species inhibit mitochondrial respiratory complex activities in cultured vascular endothelial cells Am J Physiol Cell Physiol, March 1, 2007; 292(3): C1103 - C1112. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J. Thomson, G. B. Drummond, W. S. Waring, D. J. Webb, and S. R. J. Maxwell Effects of short-term isocapnic hyperoxia and hypoxia on cardiovascular function J Appl Physiol, September 1, 2006; 101(3): 809 - 816. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A A R Thompson, J K Baillie, M Toshner, S R J Maxwell, D J Webb, and J B Irving Pericardial effusions in healthy lowlanders after acute ascent to high altitude. Heart, April 1, 2006; 92(4): 539 - 540. [Full Text] [PDF]
|
|
|
|
 |
|
 M Fluck Hypoxaemia enhanced peripheral muscle oxidative stress in COPD Thorax, October 1, 2005; 60(10): 797 - 798. [Full Text] [PDF]
|
|
|
|
 |
|
 H. Sato, M. Sato, H. Kanai, T. Uchiyama, T. Iso, Y. Ohyama, H. Sakamoto, J. Tamura, R. Nagai, and M. Kurabayashi Mitochondrial reactive oxygen species and c-Src play a critical role in hypoxic response in vascular smooth muscle cells Cardiovasc Res, September 1, 2005; 67(4): 714 - 722. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. K. Aley, K. E. Porter, J. P. Boyle, P. J. Kemp, and C. Peers Hypoxic Modulation of Ca2+ Signaling in Human Venous Endothelial Cells: MULTIPLE ROLES FOR REACTIVE OXYGEN SPECIES J. Biol. Chem., April 8, 2005; 280(14): 13349 - 13354. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Wojciak-Stothard, L. Y. F. Tsang, and S. G. Haworth Rac and Rho play opposing roles in the regulation of hypoxia/reoxygenation-induced permeability changes in pulmonary artery endothelial cells Am J Physiol Lung Cell Mol Physiol, April 1, 2005; 288(4): L749 - L760. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. B. McClelland, A. C. Dalziel, N. M. Fragoso, and C. D. Moyes Muscle remodeling in relation to blood supply: implications for seasonal changes in mitochondrial enzymes J. Exp. Biol., February 1, 2005; 208(3): 515 - 522. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-M. Li and A. M Shah Endothelial cell superoxide generation: regulation and relevance for cardiovascular pathophysiology Am J Physiol Regulatory Integrative Comp Physiol, November 1, 2004; 287(5): R1014 - R1030. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. D. Mansfield, M. C. Simon, and B. Keith Hypoxic reduction in cellular glutathione levels requires mitochondrial reactive oxygen species J Appl Physiol, October 1, 2004; 97(4): 1358 - 1366. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Chen, S. R. Thomas, A. Albano, M. P. Murphy, and J. F. Keaney Jr. Mitochondrial Function Is Required for Hydrogen Peroxide-induced Growth Factor Receptor Transactivation and Downstream Signaling J. Biol. Chem., August 13, 2004; 279(33): 35079 - 35086. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Palacios-Callender, M. Quintero, V. S. Hollis, R. J. Springett, and S. Moncada Endogenous NO regulates superoxide production at low oxygen concentrations by modifying the redox state of cytochrome c oxidase PNAS, May 18, 2004; 101(20): 7630 - 7635. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Hirayama, A. Kobiyama, S. Kinoshita, and S. Watabe The occurrence of two types of hemopexin-like protein in medaka and differences in their affinity to heme J. Exp. Biol., March 15, 2004; 207(8): 1387 - 1398. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. A. Walford, R.-L. Moussignac, A. W. Scribner, J. Loscalzo, and J. A. Leopold Hypoxia Potentiates Nitric Oxide-mediated Apoptosis in Endothelial Cells via Peroxynitrite-induced Activation of Mitochondria-dependent and -independent Pathways J. Biol. Chem., February 6, 2004; 279(6): 4425 - 4432. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Dix, T. Orth, J. Allen, J. G. Wood, and N. C. Gonzalez Activation of mast cells by systemic hypoxia, but not by local hypoxia, mediates increased leukocyte-endothelial adherence in cremaster venules J Appl Physiol, December 1, 2003; 95(6): 2495 - 2502. [Abstract] [Full Text]
|
|
|
|
|
|
S. C. Leary, C. N. Lyons, A. G. Rosenberger, J. S. Ballantyne, J. Stillman, and C. D. Moyes Fiber-type differences in muscle mitochondrial profiles Am J Physiol Regulatory Integrative Comp Physiol, October 1, 2003; 285(4): R817 - R826. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Lopes, D. Gregg, S. Vasudevan, H. Hassanain, P. Goldschmidt-Clermont, and H. Kovacic Thrombospondin 2 Regulates Cell Proliferation Induced by Rac1 Redox-Dependent Signaling Mol. Cell. Biol., August 1, 2003; 23(15): 5401 - 5408. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. O. McCarthy Rethinking Nutritional Support for Persons with Cancer Cachexia Biol Res Nurs, July 1, 2003; 5(1): 3 - 17. [Abstract] [PDF]
|
|
|
|
 |
|
 S. Turcotte, R. R. Desrosiers, and R. Beliveau HIF-1{alpha} mRNA and protein upregulation involves Rho GTPase expression during hypoxia in renal cell carcinoma J. Cell Sci., June 1, 2003; 116(11): 2247 - 2260. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Schafer, C. Schafer, N. Ewald, H.M. Piper, and Th. Noll Role of Redox Signaling in the Autonomous Proliferative Response of Endothelial Cells to Hypoxia Circ. Res., May 16, 2003; 92(9): 1010 - 1015. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Warnholtz, M. Wendt, and T. Munzel When Sleeping Beauty Turns Ugly: Mitochondria in Hypoxia Arterioscler. Thromb. Vasc. Biol., April 1, 2002; 22(4): 525 - 527. [Full Text] [PDF]
|
|
|
|
|
|
A. Warnholtz, M. Wendt, and T. Munzel When Sleeping Beauty Turns Ugly: Mitochondria in Hypoxia Arterioscler. Thromb. Vasc. Biol., April 1, 2002; 22(4): 525 - 527. [Full Text] [PDF]
|
|