[Show abstract][Hide abstract] ABSTRACT: Vascular aging is closely associated with increased vascular stiffness. It has recently been demonstrated that decreased nitric oxide (NO)-induced S-nitrosylation of tissue transglutaminase (TG2) contributes to age-related vascular stiffness. In the current study, we tested the hypothesis that exercise restores NO signaling and attenuates vascular stiffness by decreasing TG2 activity and cross-linking in an aging rat model.
Rats were subjected to 12 weeks of moderate aerobic exercise. Aging was associated with diminished phosphorylated endothelial nitric oxide synthase and phosphorylated vasodilator-stimulated phosphoprotein abundance, suggesting reduced NO signaling. TG2 cross-linking activity was significantly increased in old animals, whereas TG2 abundance remained unchanged. These alterations were attenuated in the exercise cohort. Simultaneous measurement of blood pressure and pulse wave velocity (PWV) demonstrated increased aortic stiffness in old rats, compared to young, at all values of mean arterial pressure (MAP). The PWV-MAP correlation in the old sedentary and old exercise cohorts was similar. Tensile testing of the vessels showed increased stiffness of the aorta in the old phenotype with a modest restoration of mechanical properties toward the young phenotype with exercise.
Increased vascular stiffness during aging is associated with decreased TG2 S-nitrosylation, increased TG2 cross-linking activity, and increased vascular stiffness likely the result of decreased NO bioavailability. In this study, a brief period of moderate aerobic exercise enhanced NO signaling, attenuated TG cross-linking activity, and reduced ex vivo tensile properties, but failed to reverse functional vascular stiffness in vivo, as measured by PWV.
Journal of the American Heart Association 03/2014; 3(2):e000599.
[Show abstract][Hide abstract] ABSTRACT: Nitric oxide (NO) can modulate arterial stiffness by causing regulating both functional and structural changes in the arterial wall. Tissue transglutaminase (TG2) has been shown to contribute to increased central aortic stiffness by catalyzing the crosslinking of matrix proteins. Nitric oxide (NO) S-nitrosylates and constrains TG2 to the cytosolic compartment and thereby holds its crosslinking function latent. In the present study, the role of eNOS derived NO in regulating TG2 function was studied using eNOS knockout mice. Matrix associated TG2 and TG2 cross-linking function were higher while TG2 S-nitrosylation was lower in the eNOS-/- compared to wild type (WT) mice. Pulse wave velocity (PWV) and mean arterial pressure (MAP) measured non-invasively were elevated in the eNOS-/- compared to WT mice. Intact aortas and decellularized aortic tissue scaffolds of eNOS-/- mice were significantly stiffer as determined by tensile testing. The carotid arteries of the eNOS-/- mice were also stiffer as determined by pressure-dimension analysis. Invasive methods to determine the PWV-MAP relationship showed that PWV in eNOS-/- and WT diverge at higher MAP. Thus, eNOS derived NO regulates TG2 localization and function and contributes to vascular stiffness.
AJP Heart and Circulatory Physiology 07/2013; · 4.01 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: BACKGROUND: Although ex vivo lung perfusion (EVLP) is increasingly being used to evaluate and manipulate potential donor lungs before lung transplantation (LTx), data on the biochemistry of lungs during EVLP are limited. In this study, we examined the physiology and biochemistry of human lungs on an EVLP circuit. METHODS: We recovered unallocated double lungs in standard fashion and split them into single lungs. All lungs received a nebulized arginase inhibitor, 2-S-amino-6-boronohexanoic acid (ABH), at either the onset (n = 6) or after 3 h (n = 8) of EVLP. Serial biochemical analysis included levels of arginase, endogenous nitric oxide synthase (eNOS), cyclic guanosine monophosphate, and reactive oxygen species. We considered lungs transplantable if they sustained a PaO(2):FiO(2) ≥ 350 in addition to stable pulmonary function during EVLP. RESULTS: We recovered a total of 14 single lungs. We deemed three single lungs from different donors to be transplantable after EVLP. These lungs had superior oxygenation, lower carbon dioxide, and more stable pulmonary artery pressures. Transplantable lungs had higher baseline levels of eNOS and higher final levels of cyclic guanosine monophosphate than non-transplantable lungs. Early ABH administration was associated with a transient increase in dynamic compliance. CONCLUSIONS: In this biochemical characterization of lungs deemed unsuitable for LTx, early levels of eNOS and late levels of cyclic guanosine monophosphate appear to be associated with improved allograft function during EVLP. In addition, nebulized ABH is associated with a significant increase in dynamic compliance. These data suggest that biochemical markers during EVLP may predict acceptable allograft function, and that this platform can be used to biochemically manipulate donor lungs before LTx.
Journal of Surgical Research 11/2012; · 2.12 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: OBJECTIVES: Ischemia/reperfusion injury (IRI) is a common complication of lung transplantation (LTx). Hydrogen sulfide (H(2)S) is a novel agent previously shown to slow metabolism and scavenge reactive oxygen species, potentially mitigating IRI. We hypothesized that pretreatment with inhaled H(2)S would improve graft function in an ex vivo model of LTx. METHODS: Rabbits (n = 10) were ventilated for 2 h prior to heart-lung bloc procurement. The treatment group (n = 5) inhaled room air (21% O(2)) supplemented with 150 ppm H(2)S while the control group (n = 5) inhaled room air alone. Both groups were gradually cooled to 34°C. All heart-lung blocs were then recovered and cold-stored in low-potassium dextran solution for 18 h. Following storage, the blocs were reperfused with donor rabbit blood in an ex vivo apparatus. Serial clinical parameters were assessed and serial tissue biochemistry was examined. RESULTS: Prior to heart-lung bloc procurement, rabbits pretreated with H(2)S exhibited similar oxygenation (P = 0.1), ventilation (P = 0.7), and heart rate (P = 0.5); however, treated rabbits exhibited consistently higher mean arterial blood pressures (P = 0.01). During reperfusion, lungs pretreated with H(2)S had better oxygenation (P < 0.01) and ventilation (P = 0.02), as well as lower pulmonary artery pressures (P < 0.01). Reactive oxygen species levels were lower in treated lungs during reperfusion (P = 0.01). Additionally, prior to reperfusion, treated lungs demonstrated more preserved mitochondrial cytochrome c oxidase activity (P = 0.01). CONCLUSIONS: To our knowledge, this study represents the first reported therapeutic use of inhaled H(2)S in an experimental model of LTx. After prolonged ischemia, lungs pretreated with inhaled H(2)S exhibited improved graft function during reperfusion. Donor pretreatment with inhaled H(2)S represents a potentially novel adjunct to conventional preservation techniques and merits further exploration.
Journal of Surgical Research 06/2012; · 2.12 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Ischemia-reperfusion injury is a common complication after lung transplantation. Ischemia-reperfusion injury is thought to be mediated by reactive oxygen species (ROS). Hydrogen sulfide (H(2)S) is a novel agent that has been previously shown to scavenge ROS and slow metabolism. We evaluated the effect of infused H(2)S on the presence of ROS after reperfusion in an ex vivo model of lung transplantation.
Heart-Lung blocks were recovered from New Zealand white rabbits (n = 12) and cold stored in Perfadex solution for 18 h. After storage, the heart-lung blocks were reperfused ex vivo with donor rabbit blood. In the treatment group (n = 7), a bolus of sodium H(2)S was added at the beginning of reperfusion (100 μg/kg) and continuously infused throughout the 2-h experiment (1 mg/kg/h). The vehicle group (n = 5) received an equivalent volume of saline. Serial airway and pulmonary artery pressures and arterial and venous blood gases were measured.
Oxygenation and pulmonary artery pressures were similar between the 2 groups. However, treatment with H(2)S resulted in a dramatic reduction in the presence of ROS after 2 h of reperfusion (4,851 ± 2,139 versus 235 ± 462 related fluorescence units/mg protein; P = 0.003). A trend was seen toward increased levels of cyclic guanosine monophosphate in the H(2)S-treated group (3.08 ± 1.69 versus 1.73 ± 1.41 fmol/mg tissue; P = .23).
After prolonged ischemia, infusion of H(2)S during reperfusion was associated with a significant decrease in the presence of ROS, a suspected mediator of ischemia-reperfusion injury. To our knowledge, the present study represents the first reported therapeutic use of H(2)S in an experimental model of lung transplantation.
Journal of Surgical Research 03/2012; 178(1):494-501. · 2.12 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The multifunctional enzyme tissue transglutaminase (TG2) contributes to the development and progression of several cardiovascular diseases. Extracellular rather than intracellular TG2 is enzymatically active, however, the mechanism by which it is exported out of the cell remains unknown. Nitric oxide (NO) is shown to constrain TG2 externalization in endothelial and fibroblast cells. Here, we examined the role of both exogenous and endogenous (endothelial cell-derived) NO in regulating TG2 localization in vascular cells and tissue. NO synthase inhibition in endothelial cells (ECs) using N-nitro L: -arginine methyl ester (L: -NAME) led to a time-dependent decrease in S-nitrosation and increase in externalization of TG2. Laminar shear stress led to decreased extracellular TG2 in ECs. S-nitrosoglutathione treatment led to decreased activity and externalization of TG2 in human aortic smooth muscle and fibroblast (IMR90) cells. Co-culture of these cells with ECs resulted in increased S-nitrosation and decreased externalization and activity of TG2, which was reversed by L: -NAME. Aged Fischer 344 rats had higher tissue scaffold-associated TG2 compared to young. NO regulates intracellular versus extracellular TG2 localization in vascular cells and tissue, likely via S-nitrosation. This in part, explains increased TG2 externalization and activity in aging aorta.
[Show abstract][Hide abstract] ABSTRACT: Arginase constrains endothelial nitric oxide synthase activity by competing for the common substrate, L -Arginine. We have recently shown that inducible nitric oxide synthase (NOS2) S-nitrosates and activates arginase 1 (Arg1) leading to age-associated vascular dysfunction. Here, we demonstrate that a direct interaction of Arg1 with NOS2 is necessary for its S-nitrosation. The specific domain of NOS2 that mediates this interaction is identified. Disruption of this interaction in human aortic endothelial cells prevents Arg1 S-nitrosation and activation. Thus, disruption of NOS2-Arg1 interaction may represent a therapeutic strategy to attenuate age related vascular endothelial dysfunction.
Molecular and Cellular Biochemistry 05/2011; 355(1-2):83-9. · 2.39 Impact Factor