Shear-induced reactive nitrogen species inhibit mitochondrial respiratory complex activities in cultured vascular endothelial cells.
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 microM), the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO; 100 microM), or the peroxynitrite (ONOO(-)) scavenger uric acid (UA; 50 microM), 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.
Han
Zhaosheng
Z
Department of Medicine, The Ohio State University, Columbus, OH 43210, USA.
Chen
Yeong-Renn
YR
Jones
Charles I
CI
Meenakshisundaram
Guruguhan
G
Zweier
Jay L
JL
Alevriadou
B Rita
BR
D002478:Cells, Cultured; D045222:Electron Transport Chain Complex Proteins / Q000378:metabolism*; D042783:Endothelial Cells / Q000502:physiology*; D006801:Humans; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D008928:Mitochondria / Q000378:metabolism*; D018384:Oxidative Stress / Q000502:physiology; D011312:Pressure; D026361:Reactive Nitrogen Species / Q000378:metabolism*; D033081:Shear Strength; D013314:Stress, Mechanical
D045222:Electron Transport Chain Complex Proteins; D026361:Reactive Nitrogen Species