Activation of NAD(P)H Oxidase by Tryptophan-Derived 3-Hydroxykynurenine Accelerates Endothelial Apoptosis and Dysfunction In Vivo
Rationale: The kynurenine (Kyn) pathway is the major route for tryptophan (Trp) metabolism in mammals. The Trp-Kyn pathway is reported to regulate several fundamental biological processes, including cell death.
Objective: The aim of this study is to elucidate the contributions and molecular mechanism of Trp-Kyn pathway to endothelial cell death.
Methods and Results: Endogenous reactive oxygen species (ROS), endothelial cell apoptosis, and endothelium-dependent and -independent vasorelaxation were measured in aortas of wild-type mice or mice deficient for nicotinamide adenine dinucleotide phosphate [NAD(P)H] oxidase subunits (p47phox or gp91phox) or indoleamine 2,3-dioxygenase 1 (IDO1) with or without angiotensin (Ang) II infusion. As expected, AngII increased plasma levels of Kyn and 3-hydroxykynurenine (3-OHKyn)-modified proteins in endothelial cells in vivo. Consistent with this, AngII markedly increased the expression of IDO in parallel with increased expression of interferon-gamma (IFN-γ). Further, in wild-type mice, AngII significantly increased oxidative stress, endothelial cell apoptosis, and endothelial dysfunction. These effects of AngII infusion were significantly suppressed in mice deficient for p47phox, gp91phox or IDO1, suggesting that AngII-induced enhancement of Kynurenines via NAD(P)H oxidase-derived oxidants causes endothelial cell apoptosis and dysfunction in vivo. Furthermore, IFN-γ neutralization eliminates AngII-increased superoxide products and endothelial apoptosis by inhibiting AngII-induced Kynurenines generation, suggesting that AngII-activated Kyn pathway is IFN-γ dependent. Mechanistically, we found that AngII-enhanced 3-OHKyn promoted the generation of NAD(P)H oxidase-mediated superoxide anions by increasing the translocation and membrane assembly of NAD(P)H oxidase subunits in endothelial cells (ECs), resulting in accelerated apoptosis and consequent endothelial dysfunction.
Conclusions: Kyn pathway activation accelerates apoptosis and dysfunction of the endothelium by up-regulating NAD(P)H-derived superoxide.
- indoleamine 2
- NAD(P)H oxidase,
- endothelial dysfunction
- oxidative stress
- Received June 29, 2013.
- Revision received November 24, 2013.
- Accepted November 26, 2013.