Effect of chronic angiotensin II inhibition on the nitric oxide synthase in the normal rat during aging.

OBJECTIVE: To assess the effect on the cardiovascular system, of enalapril (E) or losartan (L) given since weaning during 6 or 18 months to normal rats. METHODS: Animals were divided in three groups: control (C), E-treated and L-treated; treated rats received 10 mg/ kg per day of drug. Systolic blood pressure (SBP), body weight, water and food intake (WI, FI), cardiac, left ventricular and aortic weight as well as the length of the tail were recorded. NADPH-diaphorase activity was determined as a marker of nitric oxide synthase (NOS) activity in aorta, arterioles of small intestine, heart and kidney of normal rats. NOS activity was measured as optical density (OD) in the stained tissue. Nitrate + nitrite urinary excretion was measured in 24 h urine. Only significant differences (P < 0.05) are reported. RESULTS: SBP, absolute cardiac, left ventricular and aortic weight increased with age. Both treatments delayed these increments. At 6 and 18 months, NOS activity was higher in aortic endothelium (Em) of L- and E-treated animals. Losartan treatment during 6 months also increased NOS activity in aortic smooth muscle (SM). Aortic Em NOS activity fell in the 18 months-treated and untreated animals. E increased NOS activity in the SM of intestinal arterioles at 6 months but reduced it at 18 months. CONCLUSIONS: The fact that both E and L delayed cardiac hypertrophy/hyperplasia and aortic growth and raised aortic endothelium NOS activity indicates a protective effect on cardiovascular damage due to aging, exerted through inhibition of angiotensin II.

Effect of chronic angiotensin II inhibition on the cardiovascular system of the normal rat.

Previous studies have demonstrated in normal rats that chronic treatment, from weaning to 30 days, with either enalapril or losartan, induced significant changes in cardiovascular structure and function. The present study was performed to assess the effect of either enalapril or losartan on the structure and function of the heart and arteries given to normal rats from weaning until 6 months of age. Animals (n = 48) were divided into three groups: control, enalapril treated, and losartan treated; treated rats received 10 mg/kg/day of drug. Blood pressure, body weight, and water intake were recorded for that time period. DNA, cGMP, collagen, degree of fibrosis, and nitric oxide synthase-NADPH-diaphorase-dependent activity in the heart and arteries were determined. Only significant differences (P < .05) are reported. Blood pressure increased only in control rats (13 +/- 1 mm Hg), enalapril treatment enhanced water intake and reduced the rate of body growth (control, 672.9 +/- 15.4 g; losartan, 692.4 +/- 21.8 g; enalapril, 541.8 +/- 13.8 g). In the heart, DNA (control, 120 +/- 5; losartan, 99 +/- 4; enalapril, 93 +/- 6 microg/100 mg), collagen (control, 2.5 +/- 0.2; enalapril, 1.85 +/- 0.08 microg/100 mg), and fibrosis (control, 3.5 +/- 0.4%; losartan, 2.2 +/- 0.3%; enalapril, 2.1 +/- 0.4%) were reduced by treatment. In the aorta, cGMP (control, 0.15 +/- 0.01; losartan, 0.24 +/- 0.02 pmol/mg), and NADPH-diaphorase (control, 0.114 +/- 0.003; losartan, 0.148 +/- 0.006; enalapril, 0.169 +/- 0.003 as optical density) were enhanced. The enzyme was also higher in the aortic endothelium of treated animals (control, 0.193 +/- 0.010; losartan, 0.228 +/- 0.009; enalapril, 0.278 +/- 0.005). The lower rate of body weight increase, the enhanced water intake, and the reduced cardiac and left ventricular weight attributable to enalapril treatment do not seem to be related to inhibition of the renin-angiotensin system. On the other hand, renin-angiotensin system inhibition induces a protective effect on the heart and aorta through structural and functional changes. Most of this action seems to be exerted through angiotensin II type 1 receptors.

Effect of EXP 3174 on blood pressure of normoreninemic renal hypertensive rats.

This study examined the effect on mean blood pressure of a new orally active nonpeptide angiotensin II (Ang II) receptor antagonist, EXP 3174, in doses that completely block exogenous Ang II action. Anesthetized and conscious two-kidney, two clip chronic renovascular hypertensive rats and sham-operated animals were used. In anesthetized hypertensive rats, intracerebroventricular administration of the inhibitor had no effect on blood pressure, whereas blood pressure was normalized by intravenous injection of the antagonist (163 +/- 12 to 110 +/- 9 mm Hg, P < .05). In sham anesthetized rats, intravenous injection of EXP 3174 also lowered blood pressure (112 +/- 6 to 96 +/- 6mm Hg, P < .05). In conscious rats, intravenous EXP 3174 induced a fall in pressure that was larger in hypertensive (156 +/- 9 to 132 +/- 5 mm Hg, P < .05) than in sham (104 +/- 3 to 94 +/- 4 mm Hg, P < .05) rats. Plasma renin activity was very high in anesthetized animals (hypertensive versus sham, 87.8 +/- 8.3 versus 95.7 +/- 10.2 ng Ang I/mL per hour); differences were not significant either between anesthetized hypertensive and sham or in conscious animals (hypertensive versus sham, 9.42 +/- 1.58 versus 6.74 +/- 2.32 ng Ang I/mL per hour). Angiotensinogen concentration was higher in cerebrospinal fluid in anesthetized hypertensive rats (36.4 +/- 3.0 versus 26.0 +/- 2.4 ng Ang I/mL, P < .05) and in the artery wall of hypertensive conscious rats (103.1 +/- 10.3 versus 75.2 +/- 7.8 ng Ang I/g, P < .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Norepinephrine turnover in the heart and blood vessels of Goldblatt hypertensive rats.

The norepinephrine (NE) content, the uptake of [3H]NE, the turnover time, the turnover and the synthesis rate of the neurotransmitter in the heart and blood vessels were studied in the chronic phase of two kidney and one kidney Goldblatt renovascular hypertension, in the rat. Intact and sham operated animals were used as controls. Fifty percent of the rats subjected to renal clipping developed hypertension. This fact allowed us to compare changes in clip operated hypertensive and normotensive animals. The weight of the hearts and blood vessels was significantly increased in clip operated rats. Changes were greater in hypertensive animals. NE concentration and total content in the heart and in the artery wall were significantly decreased in the clipped rats. [3H]NE uptake was significantly diminished in the heart of experimental animals; in the artery wall, uptake was much lower than in the heart but no differences were observed between clip operated and sham animals. The turnover of NE was not different among control and clip operated rats either in the heart or in the blood vessels. Synthesis rate was lower in hypertensive animals than in their respective controls, explaining the lower concentration of the amine in cardiovascular tissues. The present data do not suggest that an increased turnover of NE in the cardiovascular sympathetic nerve endings is involved in the maintenance of high blood pressure in both types of Goldblatt renovascular hypertension.

Effect of central norepinephrine depletion on renovascular hypertension and on the renin system.

Some reports have stated that central norepinephrine (NE) depletion inhibited the development of hypertension in the rat. On the other hand, this pharmacological treatment induces changes on the central renin-angiotensin system. The present study was designed to follow the development of 2 kidney-2 clip (2k-2c) renovascular hypertension in rats depleted of central NE and to analyze the central and peripheral renin-angiotensin system. Male Wistar rats (n = 40) were used. Half of the animals was injected, intracisternally, with 6-hydroxydopamine (6-OHDA), the remaining rats only received the vehicle. One week later a silver clip was placed on each renal artery on half of the 6-OHDA treated rats and on half of the vehicle treated animals. A sham operation was performed on the remaining rats. Blood pressure was measured weekly during 7 weeks. Then, blood and cerebrospinal fluid (CSF) samples were obtained. The brain was dissected in several areas. NE and angiotensinogen concentration (AoC) were determined in tissue samples. AoC was evaluated in plasma and CSF; plasma renin activity was also measured. Hypertension development was not prevented by central NE depletion, which was significant in all central areas (p < 0.001). Other significant results showed that renal ischemia and/or NE depletion induced a significant increase in angiotensinogen concentration in the hypothalamus (p < 0.01) and in CSF (p < 0.05). In summary: central NE depletion was not able to modify the development of 2 k - 2 c hypertension. Treatment and renal ischemia induced an increase of central AoC.

Development of renovascular hypertension after central serotonin depletion.

The participation of the central serotonergic system in the development of two-kidney, two clip (2K2C) Goldblatt renovascular hypertension in the rat has been examined. Half of the rats were treated with desmethylimipramine intraperitoneally and 5,7-dihydroxytryptamine intracisternally; the other half received only desmethylimipramine and the 5,7-dihydroxytryptamine vehicle. Two days later, a silver clip was placed in both renal arteries in half of the rats of each group. A sham operation was performed in the remaining rats. Blood pressure was recorded during the 5 weeks after treatment. At the end of the experiment, blood and cerebrospinal fluid samples were obtained. The brain was dissected into several areas and kept frozen. Norepinephrine, serotonin, angiotensinogen, and renin-like concentration were evaluated in the brain areas. Plasma renin activity and angiotensinogen concentration in the plasma and cerebrospinal fluid were estimated. In the sham-operated groups, blood pressure was lower in the treated than in the control rats. The curve of blood pressure increase, as well as the final blood pressure, was similar in the treated and control 2K2C rats. Serotonin was significantly depleted by the 5,7-dihydroxytryptamine treatment in all brain areas. Treatment did not induce any changes in central norepinephrine concentration. Plasma renin activity was diminished in the treated sham-operated rats. These data indicate that the central serotonin depletion does not prevent the development of hypertension and confirm the role of the amine in normal blood pressure regulation. On the other hand, the peripheral renin-angiotensin system might participate in the development of high blood pressure in serotonin-depleted animals.

Further studies on the noradrenergic and the renin-angiotensin systems in the brain of the rat.

Previous investigations have shown that depletion of brain norepinephrine (NE) induced by chemical sympathectomy resulted in significant changes in the central renin-angiotensin system. The purpose of the present work was to increase the NE concentration in the central nervous system (CNS) in order to analyze its effect on the peptidergic complex and on the blood pressure (BP) levels. Treated rats were given the following drugs in the drinking water: 1-dopa (12 mg/rat/day), carbidopa (6 mg/rat/day) and pargyline (10 mg/rat/day) during 25 days. BP was determined, blood and cerebrospinal fluid (CSF) samples were obtained. The CNS was dissected into several areas. NE, angiotensinogen (AoC) and renin concentration (RC) were determined in the brain parenchyma; AoC was evaluated in CSF and plasma samples. Pharmacological treatment resulted in an hypotensive effect and, at the same time, an increase of NE in the CNS (about 100%; p less than 0.0005). These changes were accompanied by a significant decrease in the peripheral and central AoC. These results add new evidence to the postulated relationship between these two important regulatory systems involved in cardiovascular control.

The central and peripheral renin-angiotensin and noradrenergic systems in the spontaneous hypertensive rats (SHR).

The aim of this study was to evaluate the components of the renin-angiotensin system in the periphery and in the central nervous system (CNS) of the spontaneous hypertensive rats (SHR). On the other hand, the norepinephrine (NE) content of the different areas and of the mesenteric artery were also measured. Sixteen SHR and 9 Wistar Kyoto (WKY) control animals were used at about 6 months of age. Blood and cerebrospinal fluid (CSF) samples were collected. The brain was dissected into several areas and the mesenteric artery was excised. Plasma renin activity (PRA), plasma angiotensinogen concentration (P1AoC), brain renin (RC) and angiotensinogen concentrations (AoC) were evaluated by radioimmunoassay. NE was determined in all the tissues by a fluorimetric technique. PRA, P1AoC and NE concentration in the mesenteric artery were similar in both groups. An increase in the NE content of the cerebellum was detected in the SHR without changes in the other areas of the CNS. AoC was decreased in the CSF and in the brain stem of the SHR animals. RC was evaluated in the hypothalamus, brain stem, cerebral cortex and cerebellum of the same strain of rats. These results seem to indicate the some alteration of the peptidergic system in the CNS is present in the hypertensive animals.

Effect of central serotonin depletion on blood pressure and the renin system in rats.

In the present study we examined the effect of depletion of central nervous system serotonin by 5,7-dihydroxytryptamine on blood pressure in male Wistar rats. We also analyzed the relationship between the serotonergic and renin-angiotensin systems. Blood pressure was determined before and after intracisternal administration of 5,7-dihydroxytryptamine, 200 micrograms in saline with 1 mg/ml ascorbic acid (n = 9). Control rats (n = 8) received intracisternal vehicle. Before sacrifice, blood and cerebrospinal fluid samples were obtained. The brain was dissected in several areas. Serotonin, norepinephrine, angiotensinogen, and reninlike concentrations were determined in the brain parenchyma; angiotensinogen concentration was evaluated in cerebrospinal fluid and plasma samples; plasma renin activity was also measured. Treatment produced a significant decrease in blood pressure (-10 mm Hg; p less than 0.025) and, simultaneously, a high depletion of serotonin stores in the studied central areas (p less than 0.001), except in the cerebral cortex. Reninlike concentration was increased in the medulla oblongata (p less than 0.005) and the brainstem (p less than 0.02) after 5,7-dihydroxytryptamine treatment. Angiotensinogen concentration was decreased in the hypothalamus and elevated in the spinal cord. Angiotensinogen concentration in cerebrospinal fluid, plasma angiotensinogen concentration, and plasma renin activity did not change with treatment. Serotonin concentration in the cerebrospinal fluid remained unchanged, while the 5-hydroxyindoleacetic acid level was diminished (-47%; p less than 0.001). Intracisternal administration of 5,7-dihydroxytryptamine produced a hypotensive effect in normal rats and several modifications of the renin-angiotensin complex, suggesting a relationship between the monoaminergic and peptidergic systems.

Chronic administration of ketanserin and the development of two-kidney-two-clip Goldblatt renovascular hypertension in the rat.

The effect of ketanserin (Kt) has been analyzed during the development of two-kidney-two-clip (2k-2c) renovascular hypertension in the rat. Male Wistar rats were divided into four experimental groups: (1) clip Kt (ClKt) (n = 12)--A silver clip (0.25 mm width) was placed in each renal artery 3 days after beginning the administration of Kt (10 mg/kg/day) in the drinking water; (2) sham Kt (ShKt) (n = 13)--Similar to group 1, but the clips were placed in, and immediately removed from, the renal arteries; (3) untreated clip (UCl) (n = 10)--Similar to group 1, but the rats drank water; (4) untreated sham (USh) (n = 10)--Similar to group 2, but the rats drank water. Blood pressure (BP) was measured before surgery and was followed weekly for 7 weeks. At the end of this period, blood and cerebrospinal fluid (CSF) samples were obtained in all the animals. Plasma renin activity (PRA) and plasma and CSF angiotensinogen concentration (AoC) were evaluated. The results have shown that Kt partially inhibited the increase in BP induced by bilateral renal ischemia (BP: UCl rats 180.5 +/- 12.4 versus ClKt rats 149.8 +/- 5.1 mm Hg; p less than 0.01; USh rats 116.7 +/- 3.7; ShKt rats 114.4 +/- 5.0 mm Hg). PRA was similar in hypertensive and control rats whether or not they had received Kt. AoC in plasma was decreased in clipped treated and untreated rats.(ABSTRACT TRUNCATED AT 250 WORDS)

The brain renin-angiotensin system and the development of DOC-salt hypertension.

UNLABELLED: The effect of captopril, given in the drinking fluid, on the development of DOC-salt hypertension was analyzed. Although captopril did not prevent an increase in blood pressure (BP) elicited by DOC-salt, captopril did diminish BP in both DOC-salt and control animals. From the first week of treatment DOC-salt rats increased their fluid intake (FI). At the end of the experiment, captopril reduced this increment (655% to 357%). At the same time plasma angiotensinogen was diminished (-35%; p less than 0.001) and cerebrospinal fluid (CSF) substrate concentration increased (+33%; p less than 0.02) in DOC-salt rats, captopril did not modify these changes. In control rats captopril did not alter FI, depleted plasma angiotensinogen, (-73%; p less than 0.001), did not change the central prohormone and increased plasma renin activity (PRA) (+260%; p less than 0.001). IN CONCLUSION: CSF angiotensinogen concentration changes as previously found in CNS while a clear dissociation between plasma and CSF angiotensinogen was found in DOC-salt rats. In these animals the hypertension was not clearly affected by captopril treatment. However the effect of the converting enzyme inhibitor suggests that the central renin-angiotensin system could participate in the increase in FI.

Norepinephrine turnover in the heart and blood vessels of rats subjected to bilateral renal infarction.

The total norepinephrine (NE) content, the uptake of [3H]NE, the turnover rate and the synthesis rate of the neurotransmitter at the heart and blood vessels have been studied during the development of hypertension in rats subjected to bilateral renal infarction. Normal and sham-operated rats were used as controls. Fifty percent of the rats with renal infarction became hypertensive. The weight of the hearts and blood vessels of the experimental animals was significantly increased 15 days after renal infarction. Changes were greater in hypertensive animals. NE concentration in the heart was slightly decreased without achieving statistical significance, while total NE content was unchanged. In the artery wall NE concentration was significantly decreased in normotensive and hypertensive operated rats. [3H]NE uptake in the heart and blood vessels was similar in experimental and control animals. In relation to NE turnover, in both the heart and blood vessels, normal and sham-operated animals behaved as one population while normotensive and hypertensive rats behaved as another population. The rate constant of NE turnover was increased in both tissues of operated experimental animals without achieving statistical significance in the case of the heart. NE synthesis rate was unchanged in the cardiac muscle but was significantly increased in the blood vessels of operated animals. Present data indicate that results describing NE dynamics in the heart cannot be extrapolated for the blood vessels level; on the other hand changes in the neurotransmitter do not seem to be related to the development of high blood pressure after renal infarction in the rat.

Neonatal 6-hydroxydopamine treatment and the development of renal hypertension in the rat.

The onset and development of 2 kidney-2 clip renal hypertension was studied in chronically sympathectomized rats treated with 6-hydroxydopamine (6-OHDA) immediately after birth and with adrenal demedullation performed at the time of clipping. Blood pressure (BP) was lower in 6-OHDA treated animals than in untreated controls and the rate of hypertension development was similar in both groups. Urinary excretion of norepinephrine (NE) was significantly decreased during the 15th week and normal by the end of the 20th week. The cardiac NE content reached negligible levels while the mesenteric arteries retained 50% of its content. In the central nervous system (CNS) the 6-OHDA treatment induced a significant increase in NE concentration in the brain stem and medulla oblongata and a significant decrease in cerebellum. Hypertension produced a significant decrease in NE content in the brain stem while 6-OHDA treatment in hypertensive rats resulted in a generalized NE depletion in all the CNS areas. Results have shown that 6-OHDA treatment does not produce a complete and generally distributed sympathectomy; treatment reduces the level of BP but does not change the slope in BP increase.

Further studies on the development of two-kidney, two clip Goldblatt hypertension in 6-hydroxydopamine-treated rats.

The development of two-kidney, two clip Goldblatt hypertension (2K2C) in the rat seems to be unaffected by chemical sympathectomy with 6-hydroxydopamine (6-HODA) and adrenal demedullation. Since this treatment only partially depletes vascular norepinephrine (NE) content, we evaluated the degree of sympathectomy achieved with the 6-HODA treatment and the structural vascular changes in treated and untreated animals. Litters of male Wistar rats were divided in two groups: 6-HODA-treated (6-HODA) animals (group 1) were injected with 6-HODA since the day of birth until the end of the experiment; control group (CG) animals (group 2) received the vehicle solution. When rats reached about 250 g, a silver clip (0.25 mm width) was placed on both renal arteries in half of them in each group; a sham operation was performed on the rest of the animals. Adrenal demedullation and denervation was performed in all 6-HODA animals. Blood pressure was followed weekly by the tail cuff method for 7 weeks. At the 8th week, Silastic cannulas were placed in the carotid artery and jugular vein in all the animals. Pressor responses to tyramine (0.05 and 0.1 mg), angiotensin (10, 40 and 160 ng), and norepinephrine (NE) (25, 100 and 400 ng) and the hypotensive effect of prazosin (1 mg/kg) were determined in the conscious rats. The pressor effect of carotid occlusion was registered 24 hours later. Animals were sacrificed, and the heart and artery weight as well as the nucleic acids and alkali soluble proteins content in the vascular wall were determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Water-salt balance, plasma renin activity and catecholamine excretion in renovascular hypertension in the rat.

Water-electrolyte balance, plasma renin activity and urinary catecholamine excretion were studied for a period of 10 weeks after clipping the renal artery in the rat. Two groups of rats were examined; in Group I, a silver clip was applied on the left renal artery leaving the contralateral kidney untouched; in Group II, both renal arteries were clipped. Neither water-salt retention nor the increase inthe activity of the renin-angiotensin system or in the neural tone seem to be essential in the development of high arterial pressure after renal ischemia. All these factors would seem to be secondary mechanisms the contribution of which would depend on the experimental model or the hypertensive period under consideration.

Humoral and neurogenic factors in two-kidney renovascular hypertension.

A "bolus" dose (110 microgram) of the angiotesin II (A II)-blocker 1-Sar-8-Ala-A II (Saralasin, S) followed by its slow rate infusion (5 microgram/min/rat) for thirty min, was injected before and after the complete ganglionic blockade by pentolinium (P) in unanaesthetized unilaterally clipped renal hypertensive rats (the opposite kidney remained untouched). Pentolinium was also injected like a "bolus" dose (3 mg) followed by slow infusion (0.1 mg/min/rat) for thirty min. The observations were made until the fifth week after clipping the left renal artery. A consistent maximal hypotensive response was observed after the "bolus test" with both drugs. When S was the first drug injected, an inverse correlation was found between the percent decrease in arterial pressure (BP) by S and the percent decrease in BP by P (r = --0.83, P < 0.01, n = 8). Thus whenever a greater hypotensive effect was obtained by S, a smaller neural pressor component remained to be blocked by P. On the other hand, when P was the first drug injected a lesser A II pressor component remained to be blocked by S in the hypertensive rats. The results suggest that a considerable A II pressor effect in two-kidney renovascular hypertension is mediated via neurogenic mechanisms from the first week. A direct pressor vasoconstriction was found to be significant in cases with very high plasma-renin activity.

Vascular renin-like activity and blood pressure.

Plasma renin activity, reninlike activity present at the artery wall, pressor response to exogenous hog renin, renin half-life time, and renin-like activity present at the artery wall 1 hour after injection of renin were measured in conscious rats 1 month after inducing hypertension by renal artery constriction and contralateral nephrectomy (one-kidney hypertension). Plasma renin activity was higher but without statistical significance in one-kidney hypertensive rats when compared with normotensive or sham-operated animals. Renin-like activity present at the artery wall was significantly increased in hypertensive animals only when compared with one-kidney normotensive rats. Pressor responses to renin in one-kidney hypertensive and normotensive rats were of significantly longer duration than in sham-operated animals. The inactivation rate of exogenous renin followed a first-order reaction with a half-life of 6 minutes in sham-operated rats and of 12 minutes in one-kidney hypertensive and normotensive animals. Decreased inactivation of circulating renin could explain the protraction of the pressor response; however, the slope of the regression equation describing the inactivation of renin in all of the rats was steeper than the slope of the pressor response, indicating a dissociation between blood pressure and plasma renin activity. The renin-like activity present at the artery wall 1 hour after injection of renin was determined in the three groups; the arterial tissue of one-kidney hypertensive rats bound more circulating renin than that of normotensive rats and the latter more than that of sham-operated animals, suggesting the participation of this binding capacity in the protraction of the pressor response and in the maintenance of hypertension.