Shear-induced reactive nitrogen species inhibit mitochondrial respiratory complex activities in cultured vascular endothelial cells

• Published in: American Journal of Physiology: Cell Physiology Vol. 292; no. 3; pp. C1103 - C1112
• Main Authors: Han, Zhaosheng, Chen, Yeong-Renn, Jones, Charles I., III, Meenakshisundaram, Guruguhan, Zweier, Jay L, Alevriadou, B. Rita
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 01.03.2007
• Subjects: Blood vessels; Cell culture; Cell growth; Cells, Cultured; Electron transfer; Electron Transport Chain Complex Proteins - metabolism; Endothelial Cells - physiology; Humans; Mechanotransduction, Cellular - physiology; Mitochondria - metabolism; Nitrogen; Oxidative Stress - physiology; Pressure; Reactive Nitrogen Species - metabolism; Respiratory system; Shear Strength; Shear stress; Stress, Mechanical
• ISSN: 0363-6143; 1522-1563
• DOI: 10.1152/ajpcell.00389.2006

1 Davis Heart and Lung Research Institute, Department of Internal Medicine, and 2 Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio Submitted 14 July 2006 ; accepted in final form 30 September 2006 There is evidence that nitric oxide (NO), superoxide (O 2 – ), and their associated reactive nitrogen species (RNS) produced by vascular endothelial cells (ECs) in response to hemodynamic forces play a role in cell signaling. NO is known to impair mitochondrial respiration. We sought to determine whether exposure of human umbilical vein ECs (HUVECs) to steady laminar shear stress and the resultant NO production modulate electron transport chain (ETC) enzymatic activities. The activities of respiratory complexes I, II/III, and IV were dependent on the presence of serum and growth factor supplement in the medium. EC exposure to steady laminar shear stress (10 dyn/cm 2 ) resulted in a gradual inhibition of each of the complexes starting as early as 5 min from the flow onset and lasting up to 16 h. Ramp flow resulted in inhibition of the complexes similar to that of step flow. When ECs were sheared in the presence of the NO synthase inhibitor N G -nitro- L -arginine methyl ester ( L -NAME; 100 µM), the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO; 100 µM), or the peroxynitrite (ONOO – ) scavenger uric acid (UA; 50 µM), the flow-inhibitory effect on mitochondrial complexes was attenuated. In particular, L -NAME and UA abolished the flow effect on complex IV. Increased tyrosine nitration was observed in the mitochondria of sheared ECs, and UA blocked the shear-induced nitrotyrosine staining. In summary, shear stress induces mitochondrial RNS formation that inhibits the electron flux of the ETC at multiple sites. This may be a critical mechanism by which shear stress modulates EC signaling and function. oxidative stress; mitochondria; endothelium Address for reprint requests and other correspondence: B. R. Alevriadou, Ohio State Univ., 610 DHLRI, 473 West 12th Ave., Columbus, OH 43210 (e-mail: rita.alevriadou{at}osumc.edu )