ABSTRACT
Objective. To evaluate the impact of oxidative stress on vascular reactivity to vasoconstrictors and on nitric oxide (NO) bioavailability in saphenous vein (SV) graft with endothelial dysfunction from hypertensive patients (HT). Methods. Endothelial function, vascular reactivity, oxidative state, nitrites and NO release were studied in isolated SV rings from HT and normotensive patients (NT). Only rings with endothelial dysfunction were used. Results. HT rings presented a hyperreactivity to vasoconstrictors that was reverted by diphenylene iodonium (DPI). In NT, no effect of DPI was obtained, but Nω-nitro-(L)-arginine methyl ester (L-NAME) increased the contractile response. NO was present in SV rings without endothelial function. Nitrites were higher in NT than in HT (1066.1 ± 86.3 pmol/mg; n = 11 versus 487.8 ± 51.6; n = 23; P < 0.01) and inhibited by nNOS inhibitor. L-arginine reversed this effect. Antioxidant agents increased nitrites and NO contents only in HT. The anti-nNOS-stained area by immunohistochemistry was higher in NT than HT. HT showed an elevation of oxidative state. Conclusions. Extraendothelial NO counter-regulates contractility in SV. However, this action could be altered in hypertensive situations by an increased oxidative stress or a decreased ability of nNOS to produce NO. Further studies should be performed to evaluate the implication of these results in graft patency rates.
ABSTRACT
We investigated the effects of extraendothelial nitric oxide (NO) on angiotensin II (Ang II) reactivity in internal mammary artery (IMA) rings, as well as the impact of hypertension without associated risk factors in this response. Vascular reactivity, NO levels, and resting membrane potentials were determined in hypertensive (HT) and normotensive (NT) IMA rings. Only rings with endothelial dysfunction were included. Ang II produced a dose-dependent contraction that was higher in HT rings. Response to Ang II was potentiated by Nω-nitro L-arginine methyl ester (L-NAME) in NT but not in HT rings. The antioxidant agents tempol and diphenyleneiodonium (DPI) reverted the hyperreactivity to Ang II in HT rings. Extraendothelial NO was present in both NT and HT rings. However, NT rings showed higher values. L-NAME and S-methyl-L-thiocitrulline inhibited NO release in all cases. L-arginine reverted this inhibition. Both tempol and DPI increased NO release in both NT and HT rings. The number of vascular smooth muscle cells (VSMC) and anti-α-actin positive areas were lower in HT than in NT rings, without variations in wall thickness or wall/lumen ratio. With regard to resting membrane potential, we found in HT rings that the depolarization induced by Ang II was abolished by tempol. These findings suggest that extraendothelial NO counterregulates Ang II contractility in IMA rings; however, its action could be altered in hypertensive situations even though the patients did not have associated risk factors. We suggest two mechanisms: increased oxidative stress and a decreased ability of nNOS in VSMC to produce NO.
Subject(s)
Angiotensin II/pharmacology , Hypertension/metabolism , Mammary Arteries/drug effects , Mammary Arteries/metabolism , Muscle, Smooth, Vascular/drug effects , Nitric Oxide/metabolism , Aged , Dose-Response Relationship, Drug , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Endothelium, Vascular/physiopathology , Female , Humans , Hypertension/pathology , Hypertension/physiopathology , Male , Mammary Arteries/physiopathology , Membrane Potentials/drug effects , Membrane Potentials/physiology , Middle Aged , Muscle, Smooth, Vascular/metabolism , Muscle, Smooth, Vascular/physiopathology , NG-Nitroarginine Methyl Ester/pharmacology , Nitric Oxide Synthase Type I/metabolism , Nitrites/metabolism , Oxidative Stress/drug effects , Oxidative Stress/physiology , Vasoconstriction/drug effects , Vasoconstriction/physiologyABSTRACT
BACKGROUND: The internal mammary artery (IMA) used in bypass coronary surgery remains efficient for a longer time than other grafts, such as saphenous veins; however, its biological characteristics are incompletely defined. OBJECTIVE: To compare in IMA grafts from hypertensive (HT) and normotensive (NT) patients the presence of endothelium and their functionability, the response to passive stretching and basal tone, the reactivity to exogenous vasoconstrictors, the role of stretching in NO release, and the possible extraendothelial NO source. METHODS AND RESULTS: IMA rings contractility, presence of endothelium, and nitrite release were studied. An endothelial dysfunction associated with hypertension was found. IMA rings from HT had an impaired response to passive stretching, resulting in a decreased relaxation. All IMA grafts had an increased basal tone demonstrated by relaxation to SNP; however, a lesser response was found in HT. Interestingly, it was demonstrated that NO release was present in IMA grafts, despite an endothelial dysfunction and that stretching increased NO release. This effect was inhibited by Ca2+ -free media, L-NAME and a specific neuronal NO synthase (nNOS) inhibitor. Furthermore, the demonstration of the presence of nNOS in smooth muscle cells by immunohistochemistry supports a role of extraendothelial NO. CONCLUSION: We demonstrate the impact of hypertension in IMA grafts producing increased endothelial dysfunction, reduced response to passive stretching, increased basal tone, and impaired responsiveness to exogenous vasoconstrictors and NO release. A specific role of stretching in extraendothelial NO release was demonstrated, which may have an important role in the outcome of IMA grafts due to the protective actions of NO, even in the absence of the endothelium.
Subject(s)
Hypertension/physiopathology , Mammary Arteries/physiopathology , Nitric Oxide/physiology , Aged , Angiotensin II/pharmacology , Biomechanical Phenomena , Endothelium, Vascular/physiopathology , Enzyme Inhibitors/pharmacology , Female , Humans , In Vitro Techniques , Male , Mammary Arteries/drug effects , Middle Aged , NG-Nitroarginine Methyl Ester/pharmacology , Nitric Oxide Synthase/antagonists & inhibitors , Nitroprusside/pharmacology , Norepinephrine/pharmacology , Potassium Chloride/pharmacology , Stress, Mechanical , Vasodilation/drug effectsABSTRACT
Una estimación de la función endotelial de arterias mamarias (art mam) y venas safenas (vena saf) usadas parabypass coronario sería medir la liberación de oxido nítrico (NO). En nuestro laboratorio observamos, en arterias de conejos, que el estiramiento (stretching) incrementa el NO. Objetivos: Determinar la presencia de endotelio en art mam y vena saf en pacientes (p) de bypass coronario y dosar el contenido de NO. Evaluar el rol del estiramiento. Métodos: Se usaron restos (1-4 anillos) de art mam y vena saf de 44 p del CMC con o sin factores de riesgo (FR) asociados. Se efectuaron dos protocolos in vitro: NT) anillos no tensados (precarga de 0g) y T) anillos tensados (precarga de 2g:art mam o 3g:vena saf). La presencia de endotelio se determinó por relajación (con endotelio) o no (sin endotelio) a acetilcolina 10-6M o bradiquinina 10-7M sobre precontractura con noradrenalina en baño de órgano aislado. El NO se dosó por método de Griess a los 15 min y en una curva de tiempo (90 min) con y sin tratamiento con L-Name 10-3M o Ang II (10-9 a -6M). Resultados: 7 p presentaron endotelio sin asociación con los FR. En T el NO basal fue mayor en vasos con endotelio en art mam (1829±259 vs 3699±65 pmol/mg n=19; p<0,05) y vena saf (694±103 vs 1290±216 pmol/mg, n=18; p<0,01). T estimuló laliberación de NO, este efecto es mayor en art mam con endotelio que en vena saf (p<0,001) y se mantiene aúnen anillos sin endotelio a los 90 min. En NT no hubo diferencias entre art mam y vena saf. Ang II mostródiferencias entre T y NT. L-Name solo inhibió el NO en vena saf con endotelio. Conclusiones: Los vasospresentaron disfunción endotelial generalizada sin asociación con ningún FR. Sin embargo, hubo liberación deNO incluso en anillos sin endotelio. Demostramos mayores niveles de NO en art mam. Sin embargo art mam yvena saf aumentan su liberación si se someten a T y este aumento se mantiene en la disfunción endotelial. Suorigen involucraría activación de la enzima NO Sintetasa neuronal(nNOS). La T activaría la nNOS, situaciónhomologable in vivo a la T que sufriere el vaso injertado en el circuito arterial coronario. Este mecanismocompensador de NO ayudaría a explicar la buena perfomance, aun en vena saf, de los puentes coronarios, más allá del FR asociado. (AU)
Subject(s)
Male , Animals , Female , Nitric Oxide/metabolism , Coronary Artery Bypass/methods , Mammary Arteries , Saphenous Vein , Endothelium, Vascular/physiology , Endothelium, Vascular/physiopathology , Endothelium/physiology , Endothelium/physiopathologyABSTRACT
Una estimación de la función endotelial de arterias mamarias (art mam) y venas safenas (vena saf) usadas parabypass coronario sería medir la liberación de oxido nítrico (NO). En nuestro laboratorio observamos, en arterias de conejos, que el estiramiento (stretching) incrementa el NO. Objetivos: Determinar la presencia de endotelio en art mam y vena saf en pacientes (p) de bypass coronario y dosar el contenido de NO. Evaluar el rol del estiramiento. Métodos: Se usaron restos (1-4 anillos) de art mam y vena saf de 44 p del CMC con o sin factores de riesgo (FR) asociados. Se efectuaron dos protocolos in vitro: NT) anillos no tensados (precarga de 0g) y T) anillos tensados (precarga de 2g:art mam o 3g:vena saf). La presencia de endotelio se determinó por relajación (con endotelio) o no (sin endotelio) a acetilcolina 10-6M o bradiquinina 10-7M sobre precontractura con noradrenalina en baño de órgano aislado. El NO se dosó por método de Griess a los 15 min y en una curva de tiempo (90 min) con y sin tratamiento con L-Name 10-3M o Ang II (10-9 a -6M). Resultados: 7 p presentaron endotelio sin asociación con los FR. En T el NO basal fue mayor en vasos con endotelio en art mam (1829±259 vs 3699±65 pmol/mg n=19; p<0,05) y vena saf (694±103 vs 1290±216 pmol/mg, n=18; p<0,01). T estimuló laliberación de NO, este efecto es mayor en art mam con endotelio que en vena saf (p<0,001) y se mantiene aúnen anillos sin endotelio a los 90 min. En NT no hubo diferencias entre art mam y vena saf. Ang II mostródiferencias entre T y NT. L-Name solo inhibió el NO en vena saf con endotelio. Conclusiones: Los vasospresentaron disfunción endotelial generalizada sin asociación con ningún FR. Sin embargo, hubo liberación deNO incluso en anillos sin endotelio. Demostramos mayores niveles de NO en art mam. Sin embargo art mam yvena saf aumentan su liberación si se someten a T y este aumento se mantiene en la disfunción endotelial. Suorigen involucraría activación de la enzima NO Sintetasa neuronal(nNOS). La T activaría la nNOS, situaciónhomologable in vivo a la T que sufriere el vaso injertado en el circuito arterial coronario. Este mecanismocompensador de NO ayudaría a explicar la buena perfomance, aun en vena saf, de los puentes coronarios, más allá del FR asociado.
