Biomedical subjects

H R Weiss

Publications and source records attributed to H R Weiss.

At least 37 records · Page 2Linked to original sources

PubMed2000

Nitroprusside attenuates myocardial stunning through reduced contractile delay and time.

We hypothesized that myocardial stunning would be reversed through increased cyclic GMP caused by nitroprusside, and that this would be accomplished through a decreased proportion of regional work during diastole. Hearts were instrumented to measure left ventricular pressure, and regional myocardial mechanics were recorded using a miniature force transducer and ultrasonic dimension crystals in eight open-chest anesthetized dogs. Following baseline (CON), the left anterior descending coronary artery (LAD) was occluded for 15 min, followed by a 30-min recovery (STUN). Then intracoronary LAD infusion of sodium nitroprusside (NP) (4 microg/kg/ min) was begun. The time delay (msec) to regional shortening increased significantly from 18+/-13 to 73+/-13 following stunning, but was reduced to 49+/-18 by NP. Total regional work (g*mm/min) at baseline (1368+/-401 CON) was unchanged with stunning (1320+/-333 STUN), but reduced (961+/-240) following NP. Time to peak force development (msec) increased significantly with stunning from 284+/-13 (CON) to 333+/-11 (STUN), but was reduced to 269+/-12 following NP. The percentage work during systole was reduced from 96%+/-2% (CON) to 77%+/-7% (STUN), but returned to 98%+/-1% with NP. Regional O2 consumption was unaffected by either treatment. Cyclic GMP was unchanged by stunning (2.9+/-0.3-2.9+/-0.4 pmol/g) but increased significantly with NP (4.6+/-0.6). These data indicated that regional myocardial stunning could be attenuated by nitroprusside, which increased cyclic GMP, decreased contractile delay, increased the proportion of work done during systole, and reduced time of shortening.

Animals↗

PubMed2000

The effects of morphine on blood-brain barrier disruption caused by intracarotid injection of hyperosmolar mannitol in rats.

UNLABELLED: This study was performed to evaluate whether morphine could alter the degree of disruption of the blood-brain barrier (BBB) caused by hyperosmolar mannitol. Under isoflurane anesthesia, rats in a control group were infused with 25% mannitol into the internal carotid artery before measuring the transfer coefficient (Ki) of (14)C-alpha-aminoisobutyric acid. Infusion of morphine 3 mg/kg in the small-dose morphine group and 10 mg/kg in the large-dose morphine group was completed, 10 min before administering mannitol. There were no statistical differences in systemic blood pressures between these three groups of animals. In the control group, the Ki of the ipsilateral cortex where mannitol was injected, increased to 4.6 times that of the contralateral cortex (19.5 +/- 8.5 vs 4.2 +/- 1.2 microL. g(-1). min(-1), P < 0.002). The Ki of the ipsilateral cortex of the small-dose morphine group was 13.5 +/- 7.6 microL. g(-1). min(-1). The Ki of the ipsilateral cortex of the large-dose morphine group was 9.2 +/- 4.5 microL. g(-1). min(-1) and was smaller than that of control animals (P < 0.05). There was no significant difference in the Ki of the contralateral cortex among the three groups. In conclusion, morphine attenuated BBB disruption induced by hyperosmolar solution without significant effects on systemic blood pressure. IMPLICATIONS: Our study suggests that morphine may be effective in reducing the blood-brain barrier disruption by hyperosmolar mannitol without significant effects on systemic blood pressure.

Analgesics, Opioid↗

PubMed2000

Nitroprusside reverses lengthened time of contraction in stunned canine cardiac myocytes.

We tested the hypothesis that stunning reduces the function of isolated canine ventricular myocytes and that nitroprusside (NP) reverses this effect. After stunning (15 min occlusion, 45 min reperfusion), isolated myocytes were prepared from control (circumflex artery) and stunned (left anterior descending) regions of the hearts of seven dogs. The myocytes were examined at baseline and with NP (10(-6,-5,-4) M) for oxygen consumption (MVO2, nl O2/min/10(5) cells), cyclic guanosine monophosphate (cyclic GMP; fmol/10(5) cells), and cell contraction. Basal MVO2 was not significantly different between control and stunned myocytes (888 +/- 108 vs. 716 +/- 94). NP caused a dose dependent decrease in MVo2 (control, 262 +/-51; stunned, 287 +/- 59, NP 10(-4) M). Basal cyclic GMP levels were comparable between control and stunned myocytes (117 +/-28 vs. 124 +/- 18). NP produced a similar dose-dependent increase in cyclic GMP in control and stunned myocytes. Baseline cell shortening (%) was similar in control vs. stunned myocytes (12.1 +/- 1.2 vs. 11.0 +/- 0.9). NP reduced shortening (6.9 +/- 0.3 vs. 7.3 +/- 0.5, NP 10(-4) M). There was no baseline difference in maximal rate of shortening (microm/s) between control and stunned myocytes (164 +/- 14, 157 +/- 20). With NP, a decrease in the maximal rate of shortening was seen in both groups (128 +/- 12, 139 +/- 21, NP 10(-4) M). The time of contraction (s) was significantly longer in stunned (0.20 +/- 0.03) versus control (0.13 +/- 0.01). NP significantly lengthened the time of contraction in controls in a dose-dependent manner (0.33 +/-0.05, NP 10(-4) M). In stunned myocytes, however, low-dose NP (10(-6) M) caused a decrease in the time of contraction (0.15 +/-0.03). High-dose NP (10(-4) M) did not significantly lengthen time of contraction in stunned cells (0.23 +/- 0.02). The time of relaxation followed a similar pattern. We conclude that part of the effect of NP in low doses in stunned myocardium is to reduce the lengthened time of contraction and relaxation characteristic of stunning.

Animals↗

PubMed2000

Decreasing cyclic GMP exerts similar positive functional effects on cardiac myocytes regardless of initial level.

We tested the hypothesis that lowering the level of cyclic GMP would have positive functional effects on isolated rabbit ventricular myocytes regardless of the basal cyclic GMP level. Cell shortening data were collected with a video detector; O(2) consumption data were obtained with a Clark electrode; intracellular cyclic GMP levels were obtained by radioimmunoassay. Data were obtained: (1) at baseline; (2) after the addition of 1H-[1,2,4]oxadiazolo[4, 3-alpha]quinoxaline-1-one (ODQ) 10(-6) and 10(-4) mol/l, a selective soluble guanylyl cyclase inhibitor, and (3) after zaprinast 10(-6) mol/l, a cyclic GMP phosphodiesterase inhibitor, followed by ODQ 10(-6) and 10(-4) mol/l. We found that ODQ 10(-4) mol/l significantly decreased the cyclic GMP level from 493 +/- 75 to 301 +/- 78 (fmol/100,000 myocytes) and increased percent shortening (Pcs, %; 4.9 +/- 0.3 vs. 5.8 +/- 0.6) and maximum rate of shortening (Rs, microm/s; 58.7 +/- 5.7 vs. 73.6 +/- 4.9). Zaprinast significantly increased the cyclic GMP level from 419 +/- 140 to 599 +/- 241 and decreased Pcs (6.2 +/- 0.5 vs. 4.4 +/- 0.4) and Rs (65.5 +/- 5.3 vs. 49.6 +/- 4.3). After zaprinast, ODQ 10(-4) mol/l decreased the cyclic GMP level to 439 +/- 139 and increased percent shortening and rate of shortening by a similar percentage compared to the non-zaprinast treated myocytes. We conclude that in rabbit ventricular myocytes, a reduction in the level of myocyte cyclic GMP increases myocyte function independent of the initial cyclic GMP level.

Animals↗

PubMed1999

Effect of upregulation of AMPA glutamate receptors on cerebral O(2) consumption and blood flow in rat.

AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate) receptors, in cerebral cortex, underwent upregulation (35% increase) following chronic blockade with a non-competitive AMPA receptor antagonist, GYKI 52466 (1-(aminophenyl)-4-methyl-7, 8-methylenedioxy-5H-2,3-benzodiazepine). Such upregulation did not alter basal cerebrocortical blood flow or O(2) consumption. There was a much higher increase in blood flow and O(2) consumption in the upregulated, agonist (AMPA) stimulated cortices of anesthetized rats.

Animals↗

PubMed1999

Pacing-induced cardiac failure of hypertrophic hearts: effects of cyclic GMP reduction.

BACKGROUND: We tested the hypothesis that pacing-induced cardiac failure of hypertrophic hearts would reduce the functional and metabolic responses of these hearts to guanylate cyclase inhibition and this was associated with alterations in cyclic GMP. MATERIALS AND METHODS: Methylene blue (MB, 2 mg/kg/min, guanylate cyclase inhibitor) was infused into the left anterior descending coronary artery in 5 control, 5 left ventricular hypertrophy (LVH), and 5 LVH pacing-induced failure dogs. Regional myocardial work was calculated as the integrated product of force and segment shortening and regional myocardial O(2) consumption (VO(2)) from coronary blood flow and O(2) extraction measurements. Cyclic GMP was determined by radioimmunoassay. RESULTS: MB increased regional work (635 +/- 169 vs 1649 +/- 500, 781 +/- 184 vs 1569 +/- 203 g * mm/min) and VO(2) (8.3 +/- 1.4 vs 10.9 +/- 1.4, 7.3 +/- 0.7 vs 9.1 +/- 0.7 ml O(2)/min/100 g) in both control and LVH dogs but not in failure dogs (536 +/- 234 vs 623 +/- 193, 3.6 +/- 1.1 vs 4.7 +/- 1.9). MB also decreased cyclic GMP in control dogs (1170 +/- 142 vs 812 +/- 105 pmol/g). LVH dogs had elevated baseline cyclic GMP (5875 +/- 949) compared to control dogs but also demonstrated decreased cyclic GMP in response to MB (2820 +/- 372). In failure dogs, basal cyclic GMP was also elevated (4650 +/- 613) compared to control dogs but there was a lack of response to MB (3670 +/- 640). CONCLUSIONS: We conclude that the myocardial function, VO(2) and cyclic GMP responses to methylene blue are diminished in the transition from hypertrophy to cardiac failure.

Animals↗

PubMed1999

Effects of cyclic GMP on microvascular permeability of the cerebral cortex.

This study was performed to test the hypothesis that a direct application of cyclic guanosine monophosphate (cGMP) to the cortex would increase blood-brain barrier (BBB) permeability. Rats were anesthetized with 1.4% isoflurane and were mechanically ventilated. Two cranial windows (3 mm in diameter) were made on each side of the rat's skull (a total of four windows on each rat) to expose the cerebral cortex. A patch of normal saline, 10(-5) M, 10(-4) M, or 10(-3) M 8-bromo-cGMP was applied to each cranial window. The patches were changed every 5 min. Ten minutes after applying the patches, BBB permeability was determined by measuring the transfer coefficient (Ki) of [alpha-14C]aminoisobutyric acid. Vital signs were not changed after applying 8-bromo-cGMP. Blood gases were within normal limits. In the cortex, 10(-5) M 8-bromo-cGMP did not significantly affect the Ki; 10(-4) M 8-bromo-cGMP increased the Ki by 115%; 10(-3) M 8-bromo-cGMP increased the Ki by 124%. However, there was no statistical difference in the Ki between the doses of 10(-4) M and 10(-3) M 8-bromo-cGMP. In the pons where no patch was applied, the Ki was similar to that of the cortical area where a normal saline patch was applied. Our data demonstrated that a direct application of cGMP to the cerebral cortex significantly increased the permeability of the BBB.

Aminoisobutyric Acids↗

PubMed1999

Cyclic GMP reduces ventricular myocyte stunning after simulated ischemia-reperfusion.

We tested the hypothesis that the second messenger activated by nitric oxide, cyclic GMP, would reduce the effects of myocyte stunning following simulated ischemia-reperfusion and that this was related to cyclic GMP protein kinase. Ventricular cardiac myocytes were isolated from New Zealand White rabbits (n = 8). Cell shortening was measured by a video edge detector and protein phosphorylation was determined autoradiographically after SDS gel electrophoresis. Cell shortening data were acquired at: (i) baseline followed by 8-Bromo-cGMP 10(-6) M (8-Br-cGMP) and then KT 5823 10(-6) M (cyclic GMP protein kinase inhibitor) and (ii) simulated ischemia (20 min of 95% N(2)-5% CO(2) at 37 degrees C) followed by simulated reperfusion (reoxygenation) with addition of 8-Br-cGMP 10(-6) M followed by KT 5823 10(-6) M, (iii) addition of 8-Br-cGMP prior to ischemia followed by the addition of KT 5823 10(-6) M after 30 min of reoxygenation. In the control group, 8-Br-cGMP 10(-6) M decreased percentage shortening (%short) (5.0 +/- 0.6 vs 3.8 +/- 0. 4) and the maximum velocity (V(max), microm/s) (48.6 +/- 6.9 vs 40.2 +/- 6.4). KT 5823 10(-6) M added after 8-Br-cGMP partially restored %short (4.6 +/- 0.5) and V(max) (46.6 +/- 8.0). After stunning, baseline myocytes had decreased %short (3.4 +/- 0.2) and V(max) (36. 0 +/- 4.2). After the addition of 8-Br-cGMP, the %short (2.7 +/- 0. 2) and V(max) (27.6 +/- 2.5) decreased further. The addition of KT 5823 did not change either the %short or the V(max). The myocytes with 8-Br-cGMP during ischemia had increased %short (4.2 +/- 0.2) and V(max) (37.2 +/- 3.4) when compared to the stunned group. The addition of KT 5823 did not significantly alter %short (3.3 +/- 0.4) or V(max) (29.2 +/- 5.0) in the myocytes pretreated with 8-Br-cGMP. Protein phosphorylation was increased by 8-Br-cGMP in control and stunned myocytes. KT 5823 blocked this effect in control but not stunned myocytes, suggesting some change in the cyclic GMP protein kinase. Ischemia-reperfusion produced myocyte stunning that was reduced when 8-Br-cGMP was added prior to but not after ischemia.

Alkaloids↗

PubMed1999

A nonNMDA antagonist, GYKI 52466 improves microscopic O2 balance in the cortex during focal cerebral ischemia.

This study was performed to test whether GYKI 52466, a nonNMDA receptor antagonist, would improve microregional oxygen supply and consumption balance in the focal cerebral ischemic area. Rats were anesthetized with 1.4% isoflurane. For the GYKI Group (n = 8), 10 min before middle cerebral artery (MCA) occlusion, a bolus of 5 mg kg-1 of GYKI 52466 i.v. was administered and was followed by an infusion of 5 mg kg-1 h-1. For the Control Group (n = 8), the same volume of the vehicle was administered. One hour after MCA occlusion, regional cerebral blood flow (rCBF) was measured using the 14C-iodoantipyrine autoradiographic technique. Microscopic arterial and venous oxygen saturations were determined using microspectrophotometry. In the cortex contralateral to MCA occlusion, the average rCBF and the average O2 consumption were lower in the GYKI Group than in the Control Group (rCBF: GYKI 65.5 +/- 24.1 ml 100 g-1 min-1, Control 97.7 +/- 33.4 ml 100 g-1 min-1; O2 consumption: GYKI 3.9 +/- 1.2 ml O2 100 g-1 min-1, Control 6.2 +/- 2.5 ml O2 100 g-1 min-1) without a significant difference in the number of veins with SvO2 < 50%. In the ischemic cortex, the number of veins with SvO2 < 50% was significantly smaller in the GYKI Group (21 veins out of 63) than in the Control Group (45 out of 59) without a significant difference in the average rCBF (GYKI 44.9 +/- 17.7, Control 29.7 +/- 10.4) or regional O2 consumption between these two groups (GYKI 3.3 +/- 1.4, Control 2.7 +/- 1.2). Our data demonstrated that GYKI 52466 was effective in improving microscopic O2 balance in the focal ischemic cortical area of the brain and it decreased O2 consumption in the non-ischemic cortex.

Animals↗

PubMed1999

The effects of propranolol on heterogeneity of rat cerebral small vein oxygen saturation.

UNLABELLED: beta-Adrenergic receptors are involved in altering cerebral metabolism and blood flow. This study was performed to determine whether propranolol would alter the microregional O2 balance in the brain. Rats were anesthetized with 1.4% isoflurane. Isotonic sodium chloride solution (control group), propranolol 2 mg/kg (low propranolol group) or propranolol 20 mg/kg (high propranolol group) was administered IV to the rats. Twenty minutes later, regional cerebral blood flow (rCBF) was measured using the 14C-iodoantipyrine autoradiographic technique. Small (diameter <70 microm) arterial and venous oxygen saturation (SaO2 and SvO2, respectively) was determined using microspectrophotometry in the alternate slices of the tissue sections used to measure rCBF. In both the low and high propranolol groups, average cortical rCBF was 35% lower than that in the control group. The average O2 consumption of the cortex of the propranolol groups was significantly lower than control (low propranolol: -41%, high propranolol: -49%). In all groups, SaO2 was almost identi-cal. The heterogeneity of the microregional SvO2 expressed as the coefficient of variation (CV = 100 x sD/mean) was significantly lower in the propranolol groups (low propranolol: 8.0+/-2.3, high propranolol: 7.3 +/- 2.9) than in the control group (13.4 +/- 3.5). The proportion of cortical veins with Svo2 <55% was significantly smaller in the low and high propranolol groups (4 of 60 and 3 of 60, respectively) than that in the control group (15 of 60). In the other brain regions, the data followed a similar pattern. Our data demonstrated that propranolol is effective in decreasing O2 consumption, improving microregional O2 balance, and reducing its heterogeneity in the brain. IMPLICATIONS: Our data suggest that the linkage of O2 supply and consumption is not tightly coupled under isoflurane anesthesia. beta-Adrenergic blockers may tighten this linkage and reduce the number of low O2-saturated microregions.

Adrenergic beta-Antagonists↗

PubMed1999

Cyclic GMP protein kinase mediates negative metabolic and functional effects of cyclic GMP in control and hypertrophied rabbit cardiac myocytes.

We tested the hypothesis that in isolated cardiac myocytes, the negative metabolic and functional effects of cyclic guanosine monophosphate (GMP) are mediated by cyclic GMP protein kinase activity, and that these effects are altered in renal hypertensive (one-kidney, one-clip, 1K1C) cardiac hypertrophic rabbits. By using isolated cardiac myocytes from control and 1K1C rabbits, oxygen consumption (Mvo2; O2 nl/ min/10(5) cells), cyclic GMP (fmol/10(5) cells), and cell shortening (percentage) data were collected (a) at baseline; (b) with cyclic GMP protein kinase inhibitors KT5823 (10(-6) M) or Rp8-pCPT-cGMP (5 x 10(-6) M); (c) with the cyclic GMP phosphodiesterase inhibitor zaprinast (10(-6), 10(-4) M); and (d) with zaprinast (10(-6), 10(-4) M) and protein kinase inhibitors. Basal levels of cyclic GMP were similar in control versus 1K1C myocytes (62 +/- 10 vs. 66 +/- 17 pmol/10(5) myocytes). Zaprinast produced a dose-dependent increase in cyclic GMP in both control and 1K1C myocytes. The addition of KT5823 did not significantly affect cyclic GMP levels. Zaprinast significantly and dose dependently decreased Mvo2, and KT5823 partially restored it in control and 1K1C. Zaprinast also significantly decreased percentage shortening, and KT5823 partially restored it in control. Similar results were obtained with Rp-8pCPT-cGMP, although neither inhibitor was effective without zaprinast. The hypertrophied myocytes demonstrated comparable responses to all agents. These data suggest that the cyclic GMP protein kinase activity was not significant under basal conditions; however, the importance of cyclic GMP protein kinase in control and 1K1C myocytes was significant under conditions of increased intracellular cyclic GMP.

Alkaloids↗

PubMed1999

Effect of AMPA receptor blockade on the control of cerebral O2 supply/consumption balance in newborn pigs.

Using 1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine hydrochloride (GYKI 52466), we tested the hypothesis that alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors are important controllers of cerebral O2 supply/consumption balance in newborn piglets during both normoxia and hypoxia. Twenty-seven 2- to 7-day-old piglets were anesthetized with alpha-chloralose and were divided into four groups: 1) normoxia (n = 7), 2) GYKI 52466 (10 mg/kg, n = 7), 3) hypoxia (n = 6), and 4) hypoxia + GYKI 52466 (n = 7). We used [14C]iodoantipyrine to measure regional cerebral blood flow (rCBF) in mL/min/100 g, and we determined O2 extraction by microspectrophotometry, calculating cerebral O2 consumption (VO2) in mL O2/min/100 g in the cortex, hypothalamus, and pons. GYKI 52466 had no effect on regional VO2 or rCBF in normoxic piglets compared with controls. Hypoxia resulted in an increase in local VO2 and rCBF in the cortex and hypothalamus compared with controls: rCBF from 50 +/- 10 to 97 +/- 16 and VO2 from 2.4 +/- 0.5 to 3.7 +/- 0.4 in the cortex, and rCBF from 41 +/- 9 to 99 +/- 17 and VO2 from 2.5 +/- 1 to 3.8 +/- 0.5 in the hypothalamus. GYKI 52466 abolished this hypoxic flow effect in both the cortex (68 +/- 14) and hypothalamus (73 +/- 12). GYKI 52466 also blocked the increased VO2 in the cortex (2.5 +/- 0.4) and hypothalamus (3.0 +/- 0.5) of the hypoxic group. These findings suggest that the AMPA receptor is an important controller of VO2 in the cortex and hypothalamus during hypoxia in this immature porcine model.

Animals↗

PubMed1999

Altered effects of acetylcholine on cyclic AMP and GMP induced changes in O2 consumption of hypertrophic dog cardiac myocytes.

1. We hypothesized that acetylcholine would attenuate the metabolic effect of increasing cAMP and decreasing cGMP on cardiac myocyte O2 consumption (VO2) in dog, and this effect would be altered in left ventricular hypertrophy (LVH) produced by aortic valve placation. 2. Steady-state VO2 of a suspension of ventricular myocytes from control (n = 7) and LVH (n = 6) dogs was measured by Clark O2 electrodes during electrical stimulation (5 ms, 1 Hz, in 2 mm Ca2+). Cyclic AMP and cyclic GMP were determined by radioimmunoassay. Cellular cAMP was increased by forskolin (adenylate cyclase stimulator) and cGMP was decreased by LY83583 (guanylate cyclase inhibitor) both at 10(-7,-6,-5,-4) M with and without 10(-6) M acetylcholine. 3. Baseline cGMP level in LVH (62 +/- 10 fmol 10(-5) myocytes) was significantly greater than that in control (20 +/- 3), although the myocyte VO2 (356 +/- 39 nL O2 min(-1) 10(-5) myocytes) and cAMP levels (3.9 +/- 0.6 nmol 10(5-1) myocytes) were similar to control (312 +/- 23 and 6.9 +/- 3.1). 4. Forskolin increased myocyte cAMP in both control and LVH myocytes and increased VO2 by 51 +/- 13 in control and 91 +/- 65 in LVH myocytes. LY83583 decreased myocyte cGMP levels in control and LVH myocytes and increased VO2 by 128 +/- 57 in control and 43 +/- 26 in LVH myocytes. 5. Acetylcholine altered the cAMP, cGMP, and VO2 levels in control to 2.4 +/- 0.4, 30 +/- 3 and 213 +/- 27 and LVH to 2.5 +/- 0.3, 85 +/- 9 and 261 +/- 32. Acetylcholine attenuated the maximal effects of forskolin on VO2 to 32 +/- 27 in control and 66 +/- 56 in LVH myocytes. Acetylcholine also decreased the maximal effects of LY83583 to 82 +/- 50 in control and 19 +/- 19 in LVH myocytes. 6. The positive metabolic effects of both increases in myocyte cAMP and decreases in cGMP were blunted by acetylcholine. There was a significant increase in myocyte cGMP with forskolin in LVH myocytes. Acetylcholine decreased the increased myocyte VO2 caused by elevated cAMP or decreased cGMP in both control and LVH myocytes, although the absolute decrease in cAMP was reduced and the absolute values of cGMP were higher in LVH myocytes.

Acetylcholine↗

PubMed1999

Cyclic GMP and cyclic AMP induced changes in control and hypertrophic cardiac myocyte function interact through cyclic GMP affected cyclic-AMP phosphodiesterases.

We tested the hypothesis that the negative functional effects of cyclic GMP (cGMP) would be greater after increasing cyclic AMP (cAMP), because of the action of cGMP-affected cAMP phosphodiesterases in cardiac myocytes and that this effect would be altered in left ventricular hypertrophy (LVH) produced by aortic valve plication. Myocyte shortening data were collected using a video edge detector, and O2 consumption was measured by O2 electrodes during stimulation (5 ms, 1 Hz, in 2 mM Ca2+) from control (n = 7) and LVH (n = 7) dog ventricular myocytes. cAMP and cGMP were determined by a competitive binding assay. cAMP was increased by forskolin and milrinone (10(-6) M). cGMP was increased with zaprinast and decreased by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxilin-1-one (ODQ) both at 10(-6) and 10(-4) M, with and without forskolin or forskolin + milrinone. Zaprinast significantly decreased percent shortening in control (9 +/- 1 to 7 +/- 1%) and LVH (10 +/- 1 to 7 +/- 1%) myocytes. It increased cGMP in control (36 +/- 5 to 52 +/- 7 fmol/10(5) myocytes) and from the significantly higher baseline value in LVH (71 +/- 12 to 104 +/- 18 fmol/10(5) myocytes). ODQ increased myocyte function and decreased cGMP levels in control and LVH myocytes. Forskolin + milrinone increased cAMP levels in control (6 +/- 1 to 15 +/- 2 pmol/10(5) myocytes) and LVH (8 +/- 1 to 18 +/- 2 pmol/10(5) myocytes) myocytes, as did forskolin alone. They also significantly increased percent shortening. There were significant negative functional effects of zaprinast after forskolin + milrinone in control (15 +/- 2 to 9 +/- 1%), which were greater than zaprinast alone, and LVH (12 +/- 1 to 9 +/- 1%). This was associated with an increase in cGMP and a reduction in the increased cAMP induced by forskolin or milrinone. ODQ did not further increase function after forskolin or milrinone in control myocytes, despite lowering cGMP. However, it prevented the forskolin and milrinone induced increase in cAMP. In hypertrophy, ODQ lowered cGMP and increased function after forskolin. ODQ did not affect cAMP after forskolin and milrinone in LVH. Thus, the level of cGMP was inversely correlated with myocyte function. When cAMP levels were elevated, cGMP was still inversely correlated with myocyte function. This was, in part, related to alterations in cAMP. The interaction between cGMP and cAMP was altered in LVH myocytes.

3',5'-Cyclic-AMP Phosphodiesterases↗

PubMed1998

Exogenous nitric oxide reduces oxygen consumption of isolated ventricular myocytes less than other forms of guanylate cyclase stimulation.

We tested the hypothesis that increasing cyclic GMP with nitric oxide (NO) would reduce cardiac myocyte metabolism less than other forms of guanylate cyclase stimulation. The steady state O2 consumption (VO2) of a suspension of ventricular myocytes in 2.0 mM Ca2+ isolated from hearts of New Zealand white rabbits was measured in a glass chamber using Clark-type oxygen electrode. The cellular cyclic GMP levels, determined by radioimmunoassay, were increased by (1) adding 3-morpholinosydnonimine (SIN-1, 10(-8)-10(-5) M) and nitroprusside (10(-8)-10(-5) M), NO donors-soluble guanylate cyclase stimulators; (2) carbon monoxide (CO, 1.5 x 10(-8)-1.5 x 10(-6) M), soluble guanylate cyclase stimulator and (3) guanylin (10(-8)-10(-5) M), particulate guanylate cyclase stimulator. The baseline myocyte cyclic GMP level was 86 +/- 13 fmol/10(5) myocytes with a corresponding VO2 of 268 +/- 21 nl O2/min per 10(5) myocytes. An inverse relationship between cellular cyclic GMP levels and VO2 existed in these myocytes. The regression equations for the four treatments were: VO2 = -0.45 x [cyclic GMP] + 294.4, r = 0.94 for SIN-1; VO2 = -1.46 x [cyclic GMP] + 444.7, r = 0.96 for CO; VO2 = -1.25 x [cyclic GMP] + 389.1, r = 0.84 for guanylin and VO2 = -0.55 x [cyclic GMP] + 322.8. r = 0.79 for nitroprusside. The regression lines of the two NO donors were parallel. A similar result was also evident for the regressions of CO and guanylin. However, the slopes of both the SIN-1 and nitroprusside regression line were significantly less steep than that of either the CO or guanylin lines. Therefore, VO2 is reduced less for a similar increase in cyclic GMP with NO donors compared to direct stimulation with CO or guanylin. These results suggest that NO has metabolic effects on myocytes in addition to its stimulatory effects on cellular cyclic GMP.

Animals↗

PubMed1998

Myocardial effects of cyclic AMP phosphodiesterase inhibition are dampened in thyroxine-induced cardiac hypertrophy.

We tested the hypothesis that the increase in myocardial O2 consumption (MVO2) and myocardial wall thickening in response to milrinone would not be limited by thyroxine (T4)-induced (0.5 mg/kg for 16 days) cardiac hypertrophy. Anesthetized open-chest New Zealand white rabbits were divided into four groups: control vehicle (CV, n = 5), control milrinone (CM, n = 8), T4 vehicle (T4V, n = 7), and T4 milrinone (T4M, n = 9). Vehicle or milrinone (10(-3) M) were topically applied to the left ventricular epicardium for 15 min. Coronary blood flow (radioactive microspheres) and O2 extraction (microspectrophotometry) were used to determine O2 consumption. Cyclic AMP levels were determined by radioimmunoassay. T4 increased the heart weight to body weight ratio from 2.6 +/- 0.1 to 3.1 +/- 0.1 (g/kg). T4 rabbits had significantly higher baseline heart rates, blood pressures, and dP/dtmax and both subepicardial (EPI) and subendocardial (ENDO) blood flows. Topical application of milrinone did not have significant hemodynamic effects in either group. Baseline cyclic AMP levels (pmol/g) in the EPI and ENDO myocytes were comparable between control and T4 rabbits (CVEPI = 599 +/- 34, CVENDO = 532 +/- 26, T4VEPI = 656 +/- 42, T4VENDO = 657 +/- 17). Milrinone increased cyclic AMP in all groups although the increases were less in the T4 rabbits (CMEPI = 742 +/- 115, CMENDO = 698 +/- 101, T4MEPI = 742 +/- 103, T4MENDO = 690 +/- 55). Baseline MVO2 (ml O2/min/100 g) was significantly higher in T4 rabbits than controls (T4VEPI = 17.7 +/- 3.5 vs CVEPI = 8.5 +/- 1.5, T4VENDO = 17.2 +/- 3.2 vs CVENDO = 9.2 +/- 1.5). Significant increases in MVO2 were noted with the addition of milrinone in control (CMEPI = 14.8 +/- 3.0, CMENDO = 13.5 +/- 1.6) and T4 (T4MEPI = 25.5 +/- 3.4, T4MENDO = 22.0 +/- 3.3) rabbits; however, the percentage increase in MVO2 was significantly greater in controls (CEPI = 73%, CENDO = 47%) than T4 (T4,EPI = 44%, T4,ENDO = 28%). Thus, although the cyclic AMP phosphodiesterase activity was comparable between T4 rabbit hearts and controls, the metabolic effects and cyclic AMP effects of milrinone were dampened in this form of hypertrophy.

Animals↗

PubMed1998

Positive functional effects of milrinone and methylene blue are not additive in control and hypertrophic canine hearts.

This study was designed to test whether increased inotropy caused by raising intracellular cAMP would add to the positive inotropy caused by reducing cGMP and whether this relationship was affected by experimental hypertrophy. We used open chest anesthetized dogs with left ventricular hypertrophy (LVH) induced by valvular aortic stenosis and age matched controls (CON). Hearts were instrumented to measure local segment shortening, force, and regional work. Milrinone (MIL), a selective cyclic AMP-phosphodiesterase inhibitor, and methylene blue (MB), a guanylate cyclase inhibitor, were used to alter cAMP and cGMP levels. Ten CON and 11 LVH animals were randomly assigned to receive first either MIL (1 microg/kg/min) or MB (2 mg/kg/min) intracoronary (i.c.) infusion. After 10 min, simultaneous i.c. infusion of the other agent was begun. MIL increased regional minute work (g x mm/min) in both CON (1311 +/- 207 to 2072 +/- 285) and LVH (1052 +/- 136 to 1371 +/- 351) hearts. MB did not increase work significantly, but did increase contractile force. MIL + MB increased work from baseline; however, the combination did not increase work more than either agent alone (1961 +/- 510 CON; 1390 +/- 285 LVH). Myocardial cAMP levels (pmol/g) were significantly increased by MIL in both CON (329 +/- 53 to 437 +/- 13) and LVH hearts (351 +/- 67 to 538 +/- 32), and the addition of MB further raised cAMP (879 +/- 115 CON; 741 +/- 96 LVH). MB resulted in decreased myocardial cGMP (pmol/g) (3.20 +/- 0.61 to 2.16 +/- 0.92 CON; 5.21 +/- 1.15 to 2.46 +/- 0.56 LVH), while MIL increased cGMP (3.20 +/- 0.61 to 6.24 +/- 1.79 CON; 5.21 +/- 1.15 to 6.53 +/- 0.41 LVH). Both MIL and MB caused increases in O2 consumption, with MIL + MB together increasing O2 consumption further. The current findings demonstrate a potentiation of cAMP production in the presence of MIL + MB above either agent alone, but this did not lead to potentiation of positive functional effects. High levels of cGMP caused by milrinone may have limited the positive functional effects of cAMP.

Animals↗

PubMed1998

Altered relationship between cyclic GMP and myocardial O2 consumption in renal hypertension-induced cardiac hypertrophy.

We tested the hypothesis that preventing cyclic GMP degradation with zaprinast, (a selective cyclic GMP-phosphodiesterase inhibitor) would produce a blunted reduction in myocardial O2 consumption in renal hypertension (One Kidney-One Clip, 1K1C)-induced cardiac hypertrophy. Four groups of anesthetized open-chest New Zealand white rabbits (n = 26) were utilized. Either vehicle or zaprinast (3 x 10(-3) M) was applied topically to the left ventricular surface of control or 1K1C rabbits. Coronary blood flow (radioactive microspheres) and O2 extraction (microspectrophotometry) were used to determine O2 consumption. Myocardial cyclic GMP levels were determined by radioimmunoassay. The 1K1C rabbits had a greater heart weight-to-body weight ratio (2.94 +/- 0.08 g/kg) than controls (2.58 +/- 0.17). Systolic blood pressure was higher in 1K1C (102 +/- 9 mm Hg) than in controls (86 +/- 3). Zaprinast significantly and similarly increased cyclic GMP in both control (3.90 +/- 0.47 to 4.66 +/- 0.89 pmol/g) subepicardium (EPI) and (5.08 +/- 0.69 to 7.06 +/- 1.36) subendocardium (ENDO) and 1K1C hearts (5.53 +/- 0.61 to 7.48 +/- 1.51 EPI and 6.48 +/- 0.42 to 8.88 +/- 1.08 ENDO). Myocardial O2 consumption (ml O2/min/ 100 g) was significantly lower in controls treated with zaprinast (EPI: 8.8 +/- 0.1; ENDO: 9.5 +/- 1.9) than in controls treated with vehicle (EPI: 13.6 +/- 1.3; ENDO: 16.2 +/- 2.9). This effect was diminished in 1K1C rabbits treated with zaprinast (EPI: 10.3 +/- 2.4; ENDO: 11.2 +/- 2.6) compared with the vehicle-treated 1K1C group (EPI: 13.3 +/- 1.2; ENDO: 14.5 +/- 2.4). There was a similar increase in myocardial cyclic GMP after treatment with zaprinast, but a greater depression of myocardial O2 consumption in control animals than in 1K1C after treatment with zaprinast. This suggested that the reduction in myocardial O2 consumption, related to increases in cyclic GMP caused by cyclic GMP-phosphodiesterase blockade, was less in 1K1C cardiac hypertrophy.

Analysis of Variance↗