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Biomedical subjects

D P Nelson

Publications and source records attributed to D P Nelson.

At least 37 records · Page 2Linked to original sources
PubMed1999

Dexfenfluramine elevates systemic blood pressure by inhibiting potassium currents in vascular smooth muscle cells.

Appetite suppressants, such as dexfenfluramine (dex), are associated with primary pulmonary hypertension, valvular heart disease, and systemic vascular complications, such as coronary, cerebral, or mesenteric ischemia. These drugs suppress appetite by enhancing release and inhibiting reuptake of serotonin in the central nervous system. The effects of dex on the systemic circulation have not been studied. K(+) channels regulate vascular tone in most vascular beds. We hypothesized that dex is a systemic vasoconstrictor acting primarily by inhibiting K(+) channels, independent of effects on serotonin. The effects of clinically relevant concentrations of dex (10(-6) to 10(-4) M) on outward K(+) current and membrane potential were studied with whole-cell patch clamping in freshly isolated smooth muscle cells from rat renal, carotid, and basilar arteries. Tone was measured in tissue baths. Blood pressure, cardiac output, and left ventricular end diastolic pressure were assessed in open- and closed-chest anesthetized rats. At 10(-4) M, dex inhibits outward K(+) current (50%) and increases membrane potential (by >35 mV), an effect comparable with 4-aminopyridine (5 mM). Furthermore, dex constricts rings and acutely elevates systemic pressure (+17 +/- 3 mm Hg) and systemic vascular resistance in the presence of ketanserin. Dex vasoconstriction is dose-dependent (threshold dose 10(-6) M; 156 microg/ml) and enhanced in L-NAME-fed rats. We conclude that dex causes acute systemic vasoconstriction, at least in part by inhibition of voltage-gated K(+) channels, independent of effects on serotonin. To our knowledge, this is the first time that a commonly prescribed drug with voltage-gated K(+) channel-blocking properties is shown to have significant hemodynamic effects in vivo.

4-Aminopyridine↗
PubMed1998

Utility of a nitric oxide electrode for monitoring the administration of nitric oxide in biologic systems.

Amperometric techniques for the detection of nitric oxide (NO) are commercially available, but their sensitivity and specificity are not well described. We evaluated the sensitivity and specificity of a Clark-style, platinum NO electrode. The electrode has a lower limit of detection for NO of <25 pmol/ml in vitro and is linear over the range from 25 pmol/ml to 4 nmol/ml. The electrode is specific for NO so long as the protective membrane that covers the electrode is intact. Any defect in this membrane results in the detection of other redox agents such as hydrogen peroxide. Because of its ease of handling, specificity, and sensitivity, the NO electrode is a useful tool for quantification of administered NO in vitro and in various biologic systems.

Animals↗
PubMed1998

A role for potassium channels in smooth muscle cells and platelets in the etiology of primary pulmonary hypertension.

Plasma serotonin levels are markedly elevated in patients with primary pulmonary hypertension (PPH) and platelet levels of serotonin are low. Furthermore, plasma serotonin levels remain elevated after bilateral lung transplantation, in the absence of any pulmonary hypertension. Dexfenfluramine can cause the anorexigen-induced form of PPH that is clinically and histologically indistinguishable from PPH. We find that dexfenfluramine releases serotonin from platelets and inhibits its reuptake. These observations suggest that serotonin might be involved in, or be a marker for, the mechanism responsible for both forms of PPH. Dexfenfluramine causes inhibition of voltage-sensitive potassium (Kv) channels, membrane depolarization, and calcium entry in pulmonary artery smooth muscle cells and vasoconstriction in isolated perfused rat lungs. We have recently found that dexfenfluramine also inhibits Kv channels in megakaryocytes, the stem cell for platelets. In smooth muscle cells, taken from the pulmonary arteries of PPH patients, Kv channels appear to be dysfunctional. The underlying defect in PPH is likely to be an abnormality of one or more Kv channels in both pulmonary artery smooth muscle cells and platelets. Relatively few patients exposed to dexfenfluramine develop PPH. The factors responsible for susceptibility might be a difference in expression of potassium channels and/or a decrease in the endogenous production of nitric oxide.

Animals↗
PubMed1997

Soluble adhesion molecules in infants and children undergoing cardiopulmonary bypass.

BACKGROUND: The vascular injury and tissue damage after cardiopulmonary bypass (CPB) involves leukocyte-endothelial interactions mediated by cell adhesion molecules. This study was designed to determine the time course of soluble adhesion molecule levels after CPB in infants and children and to determine whether these levels correlated with preoperative variables, intraoperative bypass management, or postoperative course. METHODS AND RESULTS: In 56 patients undergoing CPB (median age 1.0 year, range 2 days to 19 years), plasma concentrations of soluble E-, P-, and L-selection, soluble ICAM-1, and soluble VCAM-1 were measured using sandwich enzyme-linked immunosorbent assays at the following times: at induction of anesthesia, after 15 minutes of CPB, at the end of CPB, and 1, 6, 18, and 42 hours after CPB. Preoperative, intraoperative, and postoperative data were prospectively recorded. All soluble adhesion molecule levels fell markedly at the initiation of CPB as a result of a combination of dilution and bypass circuit uptake. The time course of soluble selectins (P, E, and L), normalized to end of bypass levels, all rose significantly (P<.001) in the initial 6 hours after CPB and then returned to end bypass levels at 42 hours. Soluble ICAM-1 and VCAM rose 63% and 89% in the first 6 hours and remained elevated throughout the 42 hours. Peak soluble P-selectin levels were associated with total support time (P=.04) and preoperative cyanosis (P=.003). Soluble L-selectin levels were inversely associated with longer total support time (P=.002), longer circulatory arrest time (P=.004), longer length of intubation (P=.0009), preoperative cyanosis (P=.002), and younger age at surgery (P=.01). CONCLUSIONS: Soluble adhesion molecules have a characteristic time course in infants and children undergoing CPB. The soluble adhesion molecule levels after CPB change most significantly in patients with the highest potential for vascular injury: younger, cyanotic patients with longer pump times and longer postoperative courses. These data may be useful in the assessment of new therapies.

Adolescent↗
PubMed1997

Potassium channel diversity in vascular smooth muscle cells.

Several recent observations suggest that the vascular medium is a mosaic of functionally and morphologically unique cell types. This diversity includes differences in cell phenotype and expression of cytoskeletal and contractile proteins as well as heterogeneity of the number and activity of potassium (K+) channel types. K+ channels play a role in the regulation of arterial tone and in the control of cell proliferation. There is evidence for cell to cell, segment to segment, and vascular bed to bed diversity of K+ channels that could explain the varying responses of arterial segments or different arteries to stimuli such as hypoxia, vasoactive drugs, or arterial wall injury. Pulmonary artery vascular smooth muscle cells contain several types of K+ channels, including calcium sensitive (KCa), delayed rectifier (KDR), and ATP gated (KATP). Hypoxic pulmonary vasoconstriction (HPV) is more prominent in the resistance than in the conduit arteries. HPV is initiated by the inhibition of a KDR channel, resulting in membrane depolarization, increase in the intracellular calcium, and contraction. We have shown that some pulmonary artery smooth muscle cells are enriched in KDR channels whereas others have more KCa channels. These cells can be differentiated by their morphology (using light microscopy and electron microscopy) and their electrical properties (using patch-clamp techniques). Although present throughout the pulmonary artery, KDR-enriched cells are more prominent in the distal-resistance segments whereas KCa-enriched cells are more prominent in the proximal-conduit segments. Nitric oxide (NO) causes relaxation in part by activating a KCa channel, causing membrane hyperpolarization and inactivation of the voltage-gated calcium channels. NO is a slightly more potent vasodilator in the conduit than in the resistance pulmonary artery. In summary, the pulmonary artery may be thought of as a mosaic of cells that have different proportions of key proteins, such as K+ channel subtypes, which confer upon the cell an ability to respond to a stimulus (hypoxia or NO) differently than an adjacent cell exposed to the same stimulus. The prevalence of these cells differs from conduit to resistance arteries. Diversity of cell function may be important in physiology and pathophysiology, allowing responses to vasodilators, vasoconstrictors, and proliferative stimuli to vary within or between vascular beds.

Animals↗
PubMed1997

Induction of aquaporin-1 mRNA following cardiopulmonary bypass and reperfusion.

BACKGROUND: Cardiopulmonary bypass (CPB) and hypothermic circulatory arrest (HCA) are important components of congenital cardiac surgery. Ischemia/reperfusion injury and inflammatory cascade activation result in endothelial damage and vascular leak which are clinically manifested as pulmonary edema and low cardiac output postoperatively. Newborns are particularly susceptible. Subtraction cloning is a useful method of isolating induced genes and can be applied to CPB/HCA. MATERIALS AND METHODS: We used a newborn lamb model replicating infant CPB with HCA to obtain tissues during various periods of reperfusion. We utilized subtraction cloning to identify mRNA induced in lung following CPB/HCA and reperfusion. Ribonuclease protection was used to quantify mRNA levels. RESULTS: We isolated a cDNA encoding ovine aquaporin-1 in a subtracted cDNA screen comparing control lung with lung exposed to CPB/HCA and reperfusion. Aquaporin-1 mRNA levels increased 3-fold in lung (p = .006) exposed to CPB/HCA and 6 hr of reperfusion. No induction was observed immediately following bypass or after 3 hr of reperfusion. We found no significant induction of aquaporin-1 mRNA following bypass, arrest, and reperfusion in other tissues surveyed, including ventricle, atrium, skeletal muscle, kidney, brain, and liver. CONCLUSIONS: Our finding that aquaporin-1 mRNA is reproducibly induced in lung following CPB/HCA with 6 hr of reperfusion suggests an important role for the water channel in the setting of pulmonary edema. Induction of Aquaporin-1 is late compared with other inflammatory mediators (ICAM-1, E-selectin, IL-8). Further studies are needed to determine if aquaporin-1 contributes to the disease process or if it is part of the recovery phase.

Amino Acid Sequence↗
PubMed1996

Anorexic agents aminorex, fenfluramine, and dexfenfluramine inhibit potassium current in rat pulmonary vascular smooth muscle and cause pulmonary vasoconstriction.

BACKGROUND: The appetite suppressant aminorex fumarate is thought to have caused an epidemic of pulmonary hypertension in Europe in the 1960s. More recently, pulmonary hypertension has been described in some patients taking other amphetamine-like, anorexic agents: fenfluramine and its d-isomer, dexfenfluramine. No mechanism has been demonstrated that might account for the association between anorexic drugs and pulmonary hypertension. METHODS AND RESULTS: Using the whole-cell, patch-clamp technique, we found that aminorex, fenfluramine, and dexfenfluramine inhibit potassium current in smooth muscle cells taken from the small resistance pulmonary arteries of the rat lung. Dexfenfluramine causes reversible membrane depolarization in these cells. These actions are similar to those of hypoxia, which initiates pulmonary vasoconstriction by inhibiting a potassium current in pulmonary vascular smooth muscle. In the isolated, perfused rat lung, aminorex, fenfluramine, and dexfenfluramine induce a dose-related increase in perfusion pressure. When the production of endogenous NO is inhibited by N-nitro-L-arginine methyl ester, the pressor response to dexfenfluramine is greatly enhanced. CONCLUSIONS: These observations indicate that anorexic agents, like hypoxia, can inhibit potassium current, cause membrane depolarization, and stimulate pulmonary vasoconstriction. They suggest one mechanism that could be responsible for initiating pulmonary hypertension in susceptible individuals. It is possible that susceptibility is the result of the reduced production of an endogenous vasodilator, such as NO, but this remains speculative.

Aminorex↗
PubMed1996

Hypoxia enhances stimulus-dependent induction of E-selectin on aortic endothelial cells.

In many diseases, tissue hypoxia occurs in conjunction with other inflammatory processes. Since previous studies have demonstrated a role for leukocytes in ischemia/reperfusion injury, we hypothesized that endothelial hypoxia may "superinduce" expression of an important leukocyte adhesion molecule, E-selectin (ELAM-1, CD62E). Bovine aortic endothelial monolayers were exposed to hypoxia in the presence or absence of tumor-necrosis factor alpha (TNF-alpha) or lipopolysaccharide (LPS). Cell surface E-selectin was quantitated by whole cell ELISA or by immunoprecipitation using polyclonal anti-E-selectin sera. Endothelial mRNA levels were assessed using ribonuclease protection assays. Hypoxia alone did not induce endothelial E-selectin expression. However, enhanced induction of E-selectin was observed with the combination of hypoxia and TNF-alpha (270% increase over normoxia and TNF-alpha) or hypoxia and LPS (190% increase over normoxia and LPS). These studies revealed that a mechanism for such enhancement may be hypoxia-elicited decrements in endothelial intracellular levels of cAMP (<50% compared with normoxia). Addition of forskolin and isobutyl-methyl-xanthine during hypoxia resulted in reversal of cAMP decreases and a loss of enhanced E-selectin surface expression with the combination of TNF-alpha and hypoxia. We conclude that endothelial hypoxia may provide a novel signal for superinduction of E-selectin during states of inflammation.

1-Methyl-3-isobutylxanthine↗
PubMed1996

Blockade of selectin-mediated leukocyte adhesion improves postischemic function in lamb hearts.

BACKGROUND: Leukocyte-endothelial interactions appear to have a important role in ischemia/reperfusion injury and are mediated by specific leukocyte and endothelial adhesion molecules. The selectins are adhesion molecules found on leukocytes (L-selectin) and endothelium (P and E selectin) that bind to oligosaccharide ligands containing fucose and sialic acid to mediate leukocyte rolling on the endothelium. Fucoidin is a nontoxic sulfated fucose oligosaccharide derived from seaweed that blocks the selectins. METHODS: We tested the effects of fucoidin in an isolated blood-perfused neonatal (age range, 3 to 7 days; mean age, 4.3 days) lamb heart model undergoing 2 hours of cold cardioplegic ischemia. In group F (n = 8) fucoidin (30 mg/L) was added at initial reperfusion. Group C (n = 9) received only cardioplegia with no reperfusion intervention. Isovolumic maximum developed pressure and the maximum positive and negative first derivatives of pressure were measured using a catheter-tip transducer in an intraventricular balloon before ischemia and at 30 minutes of reperfusion. Coronary blood flow, myocardial oxygen consumption, and white blood cell counts in the circulating blood were also measured. RESULTS: Percent recoveries of baseline maximum developed pressure and maximum positive and negative first derivatives of pressure in group F (86% +/- 5%, 81% +/- 10%, and 74% +/- 8%, respectively; mean +/- standard deviation) were higher than in group C (77% +/- 5%, 70% +/- 9%, and 65% +/- 6%; p < 0.05). Group F postischemic coronary blood flow was greater (190% +/- 35%) than in group C (102% +/- 10%; p < 0.05). Recovery of myocardial oxygen consumption in group F (86% +/- 14%) was greater than group C (72% +/- 11%; p < 0.05). Postischemic white blood cell count in group F (88% +/- 4%) was greater than in group C (81% +/- 5%; p < 0.05). CONCLUSIONS: Selectin blockade with fucoidin resulted in better recovery of left ventricular function, coronary blood flow, and myocardial oxygen consumption after cold ischemia, despite a higher circulating white blood cell count. These data support the hypothesis that endothelial-leukocyte interactions play an important role in ischemia/reperfusion and suggest that selectin blockade may be a useful therapeutic strategy.

Animals↗
PubMed1996

Chronic infusion of nitric oxide in experimental pulmonary hypertension: pulmonary pressure-flow analysis.

Inhaled-nitric oxide (NO) is a selective pulmonary vasodilator in short-term clinical studies. Use of NO inhalation in chronic pulmonary hypertension is complicated by potential problems with ambulatory NO delivery. We hypothesized that long-term infusion of NO solution into the central venous circulation, which did not suffer from this drawback, might reduce chronic pulmonary hypertension. Saturated NO solution was infused in chronically hypoxic rats by implantable minipumps at a rate which was effective in reducing acute hypoxic vasoconstriction in isolated, Krebs' albumin perfused lungs (2.5 microL.h-1). Pulmonary haemo dynamics and the pressure-flow relationship were studied after 4 weeks of infusion. NO was still present in the minipumps at the end of the infusion period. Despite causing methaemoglobinaemia, NO infusion did not significantly attenuate pulmonary arterial pressure, pulmonary vascular resistance, right ventricular hypertrophy, or the parameters of the pulmonary vascular pressure-flow relationship. Pressure-flow curves, analysed with the nonlinear, distensible vessel model, indicated increased near-zero pressure resistance (Ro) in chronic hypoxia. The tendency of chronic NO infusion to reduce Ro did not reach statistical significance. Long-term infusion of nitric oxide solution is technically feasible but does not effectively reverse chronic pulmonary hypertension. The failure of infused NO to reduce pulmonary hypertension is explained by the fact that the inactivation of NO by haemoglobin is much faster than anticipated.

Animals↗
PubMed1996

The somatostatin analog angiopeptin does not reduce chronic hypoxic pulmonary hypertension in rats.

Angiopeptin is an analog of somatostatin-14, which has been found to inhibit cellular proliferation in several models of systemic vascular injury. As proliferation plays a major role in pulmonary hypertension, we examined the hypothesis that angiopeptin would inhibit the development of chronic hypoxic pulmonary hypertension in the rat. Angiopeptin was infused intravenously (90-100 microg/kg/day) by minipumps in 10 rats during a 3-week exposure to hypobaric hypoxia and in six normoxic rats. Normal saline was infused in six hypoxic control rats and in seven normoxic control rats. Angiopeptin produced no significant difference in mean pulmonary arterial pressure and resistance, right ventricular weight, or medial thickness of small pulmonary vessels. Vasoconstrictor responses of isolated lungs to acute hypoxia were not affected by angiopeptin. We conclude that angiopeptin, at the high intravenous dose used, does not significantly reduce the development of chronic hypoxic pulmonary hypertension in rats.

Animals↗
PubMed1996

Vitamin A status of children in five Ecuadorian provinces.

In mid-1993 Ecuador's Health Research Institute conducted a survey to evaluate the general nutrition and vitamin A status of children in five provinces (three in the mountains and two on the coast) containing pockets of extreme poverty. The survey enrolled 1555 children 12-59 months old who constituted a multi-phased stratified cluster sample obtained by random selection methods. Among its other aims, the survey sought to assess all or part of the study subjects' serum retinol levels, dietary vitamin A intake and ocular signs of vitamin A deficiency, and to weigh the influence on vitamin A status of age, sex, parental (maternal) education, residence in a rural or urban area, and the ethnic background of the residence area. Questionnaire interviews were conducted to gather information about each survey child's identify, diet, pathologic history, and breast-feeding history; a blood sample was obtained; and the child was weighed, measured, and given a complete physical examination (including an eye examination). Of the 1232 survey children whose serum retinol levels were measured, 18% and 2% were found to have levels below 0.7 and 0.35 mumol/L, respectively. Low serum retinol levels were more common among children of mothers who had relatively little education and resided in rural areas. The presence of Bitot's spots was confirmed in two of the study children. Interviews conducted with 39% of the study children's families to assess the children's diets showed the risk of insufficient vitamin A intake to be greater in the mountain provinces and among Indian populations, children born to mothers with no formal education, children living in rural areas, and underweight and stunted children. Forty-eight percent of the study population had serum retinol levels between 0.70 and 1.05 mumol/L, indicating marginal vitamin A deficiency. It would therefore appear that dietary supplementation would cause a substantial part of the Ecuadorian population to improve its vitamin A status. Overall, the results of the survey were consistent with a previous national survey and confirmed the existence of a pronounced subclinical vitamin A deficiency that clearly constitutes a public health problem, especially in Ecuador's rural Andean areas.

Child, Preschool↗
PubMed1996

National vitamin A survey in Panama.

The prevalence of vitamin A deficiency in a nationally representative sample of children 12-59 months old in Panama was assessed using serum retinol levels and dietary indicators. The median serum retinol level found was 1.27 +/- 0.42 mumol/L (38 micrograms/dL); 6.0% of the study sample providing adequate blood specimens had levels below 0.7 mumol/L (20 micrograms/dL), indicating deficient vitamin A intake. The Panama City Metropolitan Area and the country's western region had the highest prevalences of low serum retinol levels (below 0.7 mumol/L in 9% and 6% of the study children, respectively), as compared to overall prevalences of 5% in the two other regions studied. Low serum retinol levels were significantly more prevalent among Indians in the study group (primarily Guaymí Indians) than among non-Indians (13% versus 5%). Dietary information provided by the study children's mothers showed that high risk of inadequate dietary vitamin A intake closely paralleled low serum retinol levels; specifically, the highest prevalence of dietary inadequacy was found in the western region, especially among the Indians. The Panamanian Government is currently increasing distribution of high-dose vitamin A capsules to Indian preschoolers in Chiriquí and Bocas del Toro Provinces.

Child Nutrition Disorders↗
PubMed1995

Opposing effects of oxidants and antioxidants on K+ channel activity and tone in rat vascular tissue.

K+ channels regulate tone in both the systemic and pulmonary circulations. K+ channel inhibition leads to membrane depolarization, Ca2+ influx and vasoconstriction; K+ channel activation leads to hyperpolarization and vasodilatation. The sulfhydryl oxidant diamide opens K+ channels in pulmonary smooth muscle and acts as a potent vasodilator in perfused lungs. We examined the hypothesis that antioxidants cause constriction and oxidants cause relaxation through their effects on K+ channels in vascular smooth muscle. The oxidant diamide (380 microM and 3.8 mM) inhibited the reduction of cytochrome C by ferrous sulphate in vitro whilst the antioxidants co-enzyme Q10 (770 microM) and duroquinone (700 microM) increased the rate of reduction. Both antioxidants caused dose-dependent constriction of endothelium-intact and -denuded rat pulmonary artery and aortic rings. This constriction could be reversed by 1 microM diamide. Co-enzyme Q10 and duroquinone (both at 100 microM) partially inhibited (approximately 30%) whole-cell K+ channel currents and depolarized membranes of isolated pulmonary artery smooth muscle cell recorded using the amphotericin-perforated-patch-clamp technique. Diamide (100 microM) increased whole-cell K+ channel currents and hyperpolarized the membrane. The data suggest that oxidants and antioxidants may modulate vascular tone via an effect on K+ channels.

Animals↗
PubMed1995

Dithionite increases radical formation and decreases vasoconstriction in the lung. Evidence that dithionite does not mimic alveolar hypoxia.

Dithionite is a powerful reducing agent used to deoxygenate hemoglobin and create anaerobic conditions in vitro. Recently, dithionite has been used as a convenient means of creating "hypoxia" in experiments studying the O2 sensor in the pulmonary circulation and carotid body. We evaluated the hypothesis that hypoxia created by hypoxic ventilation and that created by dithionite have different effects on the pulmonary circulation. In vitro, dithionite (10(-5) to 10(-3) mol/L), added to oxygenated Krebs' solution, rapidly created superoxide anion in a dose-dependent manner. Dithionite consumed O2 in parallel with the generation of superoxide radical, with both processes peaking within seconds. Anoxia was sustained only if resupply of O2 was prevented. In isolated rat lungs (whether perfused with autologous blood or Krebs' solution), hypoxic ventilation alone lowered perfusate PO2 from approximately 140 to 40 mm Hg and decreased lung levels of activated oxygen species (AOS), measured by luminol-enhanced chemiluminescence, before the onset of hypoxic pulmonary vasoconstriction. Constrictor responses to angiotensin II and KCl were not impaired by intermittent hypoxic challenges, and lung weight did not increase. In contrast, dithionite impaired constrictor responses of the Krebs' solution-perfused lungs to all vasoconstrictors tested and increased lung weight. When given as a bolus (5 x 10(-3) mol/L) into the pulmonary artery during normoxic ventilation, dithionite caused no vasoconstriction and only briefly lowered PO2 (because of constant resupply of O2 from the alveoli). When superimposed on hypoxic ventilation, dithionite further lowered PO2 from approximately 40 to approximately 0 mm Hg and caused additional constriction. Unlike hypoxic ventilation, dithionite increased AOS production. Antioxidant enzymes diminished dithionite-induced radical production and diminished the loss of vascular reactivity and lung edema. In conclusion, unlike hypoxic ventilation, dithionite causes edema and loss of vascular reactivity in the lung by generating superoxide anion and hydrogen peroxide. Hypoxia elicited by dithionite is not equivalent to authentic hypoxia because of the obligatory associated generation of AOS. Dithionite usage should not be substituted for authentic hypoxia in studies of O2 sensing.

Animals↗
PubMed1994

Nitric oxide and cGMP cause vasorelaxation by activation of a charybdotoxin-sensitive K channel by cGMP-dependent protein kinase.

Nitric oxide (NO)-induced relaxation is associated with increased levels of cGMP in vascular smooth muscle cells. However, the mechanism by which cGMP causes relaxation is unknown. This study tested the hypothesis that activation of Ca-sensitive K (KCa) channels, mediated by a cGMP-dependent protein kinase, is responsible for the relaxation occurring in response to cGMP. In rat pulmonary artery rings, cGMP-dependent, but not cGMP-independent, relaxation was inhibited by tetraethylammonium, a classical K-channel blocker, and charybdotoxin, an inhibitor of KCa channels. Increasing extracellular K concentration also inhibited cGMP-dependent relaxation, without reducing vascular smooth muscle cGMP levels. In whole-cell patch-clamp experiments, NO and cGMP increased whole-cell K current by activating KCa channels. This effect was mimicked by intracellular administration of (Sp)-guanosine cyclic 3',5'-phosphorothioate, a preferential cGMP-dependent protein kinase activator. Okadaic acid, a phosphatase inhibitor, enhanced whole-cell K current, consistent with an important role for channel phosphorylation in the activation of NO-responsive KCa channels. Thus NO and cGMP relax vascular smooth muscle by a cGMP-dependent protein kinase-dependent activation of K channels. This suggests that the final common pathway shared by NO and the nitrovasodilators is cGMP-dependent K-channel activation.

Animals↗
PubMed1994

Increased endothelium-derived NO in hypertensive pulmonary circulation of chronically hypoxic rats.

The hypothesis that the endothelium-derived relaxing factor/nitric oxide (EDNO) activity is elevated in chronic hypoxic pulmonary hypertension (CH-PHT) was tested using isolated Krebs-albumin-perfused rat lungs. Concentration of the EDNO decomposition products (NOx) in the lungs' effluent was measured by a modified chemiluminescence assay. The functional significance of basal EDNO production was studied by measuring the vasoconstrictor response to an EDNO synthesis inhibitor, N omega-nitro-L-arginine methyl ester (L-NAME). Reactivity to the endothelium-dependent vasodilator substance P and to exogenous NO was also studied. More NOx was found in effluent from CH-PHT (22.3 +/- 9.8 nM) than control (0.4 +/- 3.9 nM) lungs. The L-NAME-induced vasoconstriction was greater in CH-PHT than in control rats. The sensitivity, but not the maximal vasodilation, to exogenous NO was elevated in CH-PHT. The substance P-induced vasodilation was potentiated in CH-PHT compared with control rats and blocked by L-NAME in both groups. We conclude that basal and agonist-stimulated pulmonary EDNO activity is enhanced in this model of CH-PHT. The EDNO synthesis may play a counterregulatory role in CH-PHT.

Animals↗