Renal nerves participation in the effects of nitric oxide and ET(A)/ET(B) receptor inhibition in spontaneously hypertensive rats.

The influence of renal nerves on the effects of concurrent NO synthase inhibition (10 mg kg(-1) b.w. i.v. L-NAME) and ET(A)/ET(B) receptor inhibition (10 mg kg(-1) b.w. i.v. bosentan) on renal excretory function and blood pressure in conscious spontaneously hypertensive rats (SHR) was investigated. L-NAME increased blood pressure, urine flow rate, fractional excretion of sodium, chloride and phosphate in both normotensive Wistar rats and SHR with intact renal nerves (p<0.01). GFR or RBF did not change in any of the groups investigated. The effects of L-NAME on renal excretory function were markedly reduced by bosentan and the values returned to control level in the normotensive rats, while in SHR the values were reduced by bosentan, but they remained significantly elevated as compared to control level (p<0.05). The hypertensive response induced by L-NAME in SHR is partially due to activation of endogenous endothelins, but it does not depend on renal nerves. Chronic bilateral renal denervation abolished the effect of L-NAME on sodium and chloride excretion in normotensive rats, whereas it did not alter this effect in SHR. The participation of endogenous endothelins in changes of renal excretory function following NO synthase inhibition is diminished in SHR as compared to Wistar rats.

Renal effects of acute nitric oxide and ET(A)/ET(B) receptor inhibition in conscious spontaneously hypertensive rats.

The aim of the present study was to investigate the effects of endogenous endothelin on renal excretory function in spontaneously hypertensive rats (SHR) after inhibition of NO synthesis. The effects of non-selective ET(A)/ET(B) receptor blockade on L-NAME-induced changes in renal excretory function and blood pressure (BP) were investigated in conscious, SHR and normotensive Wistar rats with implanted catheters in the bladder for urine collection, in the femoral artery for BP registration and in the femoral vein for L-NAME and bosentan administration. L-NAME increased systolic, mean and diastolic BP, diuresis, sodium and chloride excretion (p < 0.01) in both normotensive and hypertensive rats but bosentan returned the values of diuresis, sodium and chloride excretion to control level without any changes in BP in normotensive rats. In SHR the effects of L-NAME were reduced after bosentan (p < 0.05) but the values of diuresis, sodium and chloride excretion still remained statistically significant as compared to control level despite the fact that bosentan lowered mean and diastolic BP increased due to L-NAME administration. Endogenous endothelins participate in a different manner in the rise of BP and in the changes in renal excretory function that develops after L-NAME-induced NO synthase inhibition in normotensive rats and in SHR.

Influence of renal denervation on renal effects of acute nitric oxide and ETA/ETB receptor inhibition in conscious normotensive rats.

The role of renal nerves in the effects of concomitant NO synthase and non-selective ET(A/)ET(B) receptor inhibition on renal function was investigated in conscious normotensive Wistar rats. NO synthase inhibition alone (10 mg/kg b. w. i.v. L-NAME) in sham-operated rats with intact renal nerves induced an increase in systolic, diastolic and mean arterial pressure, urine flow rate, sodium, chloride and calcium excretion (p<0.05). The effect of L-NAME was markedly reduced by bosentan (10 mg/kg b.w. i.v.) and the values of urine flow rate, sodium, chloride and calcium excretions returned to control level (p<0.05). L-NAME administration one week after a bilateral renal denervation increased blood pressure to a similar extent as in sham-operated rats but decreased urine flow rate (p<0.05) and did not change electrolyte excretion. ET(A/)ET(B) receptor inhibition with bosentan during NO synthase inhibition in the renal denervated rats did not produce changes in urine flow rate or electrolyte excretion. NO synthase inhibition as well as concurrent NO synthase and ET(A/)ET(B) receptor inhibition did not change clearance of inulin or paraaminohippuric acid in sham-operated or renal denervated rats. These results indicate that renal sympathetic nerves play an important modulatory role in NO and endothelin induced effects on renal excretory function.

Renal endothelin system and excretory function in Wistar-Kyoto and Long-Evans rats.

AIM: The role of the kidney endothelin system in the renal regulation of fluid and electrolyte excretion was investigated in Wistar-Kyoto (WKY) and Long-Evans (LE) rats in which we found previously marked differences in the renal excretory responses to endothelin A receptor blockade. METHODS: The selective endothelin A and B receptor antagonists BQ-123 (16.4 nmol kg(-1) min(-1)) and BQ-788 (25 nmol kg(-1) min(-1)) were infused i.v. for 50 min in conscious chronically instrumented WKY and LE rats and their renal function and renal endothelin system were studied. RESULTS: Without effects on glomerular filtration rate or renal blood flow, BQ-123 and BQ-788 decreased by more than 50% (P < 0.01) both urine flow rate and electrolyte excretion in WKY rats but only urine flow rate (P < 0.05) in LE rats. Endothelin-1 content, preproET-1/GPDH mRNA ratio, B(max) and K(d) of total endothelin receptors in renal cortex did not differ between the two strains. In contrast, plasma endothelin-1 concentration (0.58 +/- 0.04 vs. 1.05 +/- 0.01 femtomol mL(-1); P < 0.01), renal papillary ET-1 concentration (68 +/- 5 vs. 478 +/- 62 fmol mg(-1) protein; P < 0.01) and preproET-1/GPDH mRNA ratio (0.65 +/- 0.09 vs. 0.88 +/- 0.05; P < 0.05) as well as total endothelin receptor number in renal papilla (B(max) 5.3 +/- 0.4 vs. and 9.0 +/- 1.2 pmol mg(-1) protein; P < 0.05) were markedly lower in LE than in WKY rats. In vitro studies showed that in both strains ET(B) receptors on renal cortical membranes amounted between 65% and 67% and on papillary membranes between 85% and 88%. CONCLUSION: The present data show that the selective ET(A) or ET(B) receptor blockade differentially affects tubular water and salt handling, which becomes apparent in conditions of low renal papillary endothelin receptor number and tissue endothelin-1 concentration.

Influence of nonselective ET(A)/ET(B) receptor blockade on renal function in conscious rats: effects of renal denervation.

The effects of nonselective ET(A)/ET(B) receptor blockade with intravenous bolus injection of bosentan (10 mg/kg) on renal excretory function and blood pressure were investigated in conscious, male, normotensive Wistar rats before and one week after bilateral renal denervation. Renal denervation was followed by an increase in urine flow rate from 4.54+/-0.38 to 5.72+/-0.36 microl/min x 100 g b.w. (p<0.05) and a decrease in urine osmolality from 855.5+/-44.6 to 707.4+/-47.5 mosm/kg H(2)O (p<0.05). Bosentan administration in sham-operated rats resulted in decrease in urine flow rate from 4.54+/-0.38 to 3.49+/-0.34 microl/min x 100 g b.w. (p<0.05), and increase in urine osmolality from 855.5+/-44.6 to 1075.0+/-76.1 mosm/kg H(2)O (p<0.05). Sodium excretion decreased from 226.9+/-20.0 to 155.1+/-11.0 nmol/min x 100 g b.w. (p<0.01). Bosentan administration in renal denervated rats did not produce any changes in renal water or electrolyte excrections. Blood pressure, heart rate, clearance of Inulin or clearance of paraaminohippuric acid (PAH) did not change in sham-operated or renal denervated rats during nonselective ET(A)/ET(B) receptor blockade. Bosentan did not alter the baroreflex sensitivity or sympatho-vagal balance in sham-operated or renal denervated rats. In conclusion, an interaction between renal nerves and endothelins appears to be involved in the regulation of the renal excretory function.

Blood pressure variability in conscious spontaneously hypertensive rats during endothelin A receptor inhibition.

Spontaneously hypertensive rats (SHR) and normotensive Wistar rats were treated with the endothelin(A) (ET(A)) receptor antagonist BQ-123 16.4 nmol/kg.min i.v. at an infusion rate of 25 microl/min for 50 min. Blood pressure was measured in the femoral artery with a Gould/Statham P23ID transducer connected to the computerized data acquisition system Biopac MP100WS. Values of interpulse interval, systolic, diastolic and mean arterial pressure were determined in every heart cycle. Spectrum analysis of systolic, diastolic and mean arterial pressure and interpulse interval variabilities was performed using fast Fourier transform algorithm. In contrast to normotensive rats, BQ-123 infusion in SHR increased mid-frequency band in systolic, diastolic and mean arterial pressure (p < 0.05). The baroreflex sensitivity obtained by the alpha-coefficient in the mid-frequency region (square root of the relation PMFIPI/PMFSAP in msec/mmHg) did not differ between Wistar and SHR before and during BQ-123 administration. Endogenous endothelin acting via endothelin(A) receptors modulates the effects of the sympathetic nervous system on the variations of systolic, diastolic and mean arterial pressure in the hypertensive state. This interrelation between endogenous endothelin and the sympathetic nervous system is not dependent on the arterial baroreflex and is most likely to be realized locally on the vessel level.

Involvement of renal nerves and endothelins in the regulation of renal water excretion in diabetes insipidus rats.

The interaction between renal nerves, endothelins acting via endothelin-A receptors and vasopressin in the regulation of renal excretory function was investigated. In conscious intact and renal denervated diabetes insipidus (DI) Brattleboro rats, as well as their controls, Long-Evans (LE) rats, an infusion of 16.4 nmol/kg/min ET(A) receptor antagonist BQ-123 was performed in the course of 50 min. Femoral artery blood pressure, heart rate, Ccr, V x U(Na), V x U(K) and V x U(Cl) did not alter in any of the groups. Urine flow rate diminished by 38.1% (p < 0.02), while urine osmolality increased by 30.3% (p < 0.05) as a result of BQ-123 infusion in the intact LE rats but neither urine flow rate nor urine osmolality changed in the DI rats. In contrast to intact LE rats, BQ-123 infusion in renal denervated LE rats did not alter urine flow rate or urine osmolality. However, urine flow rate in renal denervated DI rats surprisingly decreased by 71.1% (p < 0.01) while urine osmolality increased by 161% (p < 0.001) as a result of BQ-123 infusion. Endogenous endothelins can regulate renal water excretion through ET(A) receptor activation. Renal sympathetic nerves participate in the modulation of renal water excretion influencing the ET(A) receptor-mediated effects of endothelins in the kidney.

Renal nerves and endothelins interaction in the control of renal excretory function in conscious Long-Evans rats.

The role of renal nerves and endothelins, acting at ET(A) receptors, in the regulation of renal excretory function was investigated in male Long-Evans rats. Catheters were placed in the femoral vein for fluid and drug infusion, in the femoral artery for blood pressure recording as well as in the bladder for urine collection. Infusion of 16.4 nmol/kg/min of the ET(A) receptor antagonist BQ-123 for 50 min was performed in freely moving, intact and renal denervated rats. As a result of BQ- 123 infusion, urine flow rate diminished (P < 0.02) and Uosm increased (P < 0.05) in the intact rats, but not in the renal denervated rats. Bilateral renal denervation itself as well as ET(A) receptor inhibition in both intact and renal denervated rats did not change the mean arterial pressure, heart rate, or the excretion of sodium, potassium and chloride. The data obtained suggest an interrelationship between renal nerves and endothelin-A receptors in the regulation of renal water excretion.

Changes of renal function and blood pressure after nitric oxide synthase inhibition in renal-denervated conscious rats.

Conscious male Wistar rats with implanted catheters in femoral artery for blood pressure registration, femoral vein for drug infusion and bladder for urine collection were used. Nitric oxide synthase inhibition (NOSI) was performed by intravenous administration of 10 mg/kg b.w. N(omega)-nitro-L-arginine methyl ester (L-NAME) before and one week after bilateral renal denervation. Renal denervation led to a decrease of urine osmolality (p<0.05). NOSI decreased heart rate (p<0.001) and increased systolic, mean and diastolic artery pressure both in the intact and the renal-denervated rats (p<0.001). The P(MF)/P(HF) ratio in the heart rate spectrum, considered a criterion for the sympathovagal balance decreased after NOSI in intact and in renal-denervated rats (p<0.001) indicating a reduction in the sympathetic tone. The baroreflex sensitivity after NOSI increased both in conscious rats with intact renal nerves and in rats with bilateral renal denervation. In intact rats NOSI increased urine flow rate by 48.7% (p<0.05), sodium excretion by 339.7% (p<0.01) and chloride excretion by 272.1% (p<0.01). In contrast to the intact rats, NOSI in the renal denervated rats decreased their urine flow rate by 35.5% (p<0.05) and did not alter sodium and chloride excretion. So, chronic renal denervation inversed the diuretic and clearly attenuated the natriuretic and chloruretic response to acute NOSI without inhibiting the rise in blood pressure. Therefore, the increased urine flow as well as the sodium and chloride excretion observed after NOSI are not exclusively the result of a pressure diuresis but are somewhat dependent on the renal sympathetic nerve activity. Renal denervation did not change the pattern of sympathovagal balance and baroreflex sensitivity modified by NO-synthase inhibition.

Spectral coherence between blood pressure and inter-beat intervals in hypertension.

Continuous registration of blood pressure (BP) was obtained in 31 normotensives and 76 hypertensives (divided into 3 stages of hypertension) by a method where the blood pressure in a human finger was measured non-invasively in a beat-by-beat fully calibrated manner. From this signal, the power density spectra of inter-beat intervals (IBI), systolic blood pressure (SBP) and diastolic blood pressure (DBP) were derived. Total spectral power (TP) and low/high spectral frequency ratios of all spectra were calculated. Cross-correlation analysis between IBI, SBP and DBP spectra showed that the level of amplitude and phase spectral coherence (SC) was strongly dependent of the stage of hypertension. SC was calculated by means of appropriate mathematical algorithms. Multiple step regression analysis pointed out that the observed phenomenon is highly significant for both--the amplitude and the phase SC (p < 0.01).

Renal excretory function in conscious Long Evans and vasopressin deficient (Brattleboro) rats after endothelin-A receptor inhibition.

All experiments were performed on conscious, freely moving male Long Evans as well as Diabetes incipidus (Brattleboro) rats (300-320 g). The endothelin-A (ETA) receptor antagonist BQ-123 (Neosystem) was administered through femoral vein cannula. Arterial blood pressure was measured trough femoral artery catheter. The bladder was cannulated for urine collection via a small suprapubic incision. After a 40 min control period BQ-123 infusion (16.4 nmol/kg/min, 25 microliters/min) was started and continued for 50 min. The effect of 32.8 nmol/kg/min BQ-123 infused in conscious Brattleboro rats was also investigated. Plasma and urine concentrations of sodium, potassium and chloride as well as osmolality were determined. Glomerular filtration rate (GFR) was estimated using the clearance of endogenous creatinine. Endothelin-A receptor inhibition by 16.4 nmol/kg/min BQ-123 infusion in conscious Long-Evans rats decreased urine flow rate by 38.4% (p < 0.02) and increased urine osmolality by 30.3% (p < 0.05). Sodium, potassium, chloride excretion did not alter. Endothelin-A receptor inhibition by 16.4 nmol/kg/min and by 32.8 nmol/kg/min BQ-123 infusion in conscious Brattleboro rats did not produce any change in urine flow rate, urine osmolality or excretion of the electrolytes studied. Endothelins acting via ETA receptors may function as an inhibitor of water reabsorption in the kidneys of conscious rats.

Spectral analysis of heart rate and arterial pressure variability after nitric oxide synthase inhibition.

The experiments were performed on male, conscious Wistar rats. Femoral arterial pressure was registered by Statham GOULD P23 ID pressure transducer connected to MP 100WS BIOPAC work station after analog to digital conversion during 40 minutes long control period. Nitric oxide synthase inhibition was performed by injection of 100 microliters, 10 mg/kg b.w. N-omega-nitro-L-arginine methyl ester (L-NAME) in saline through femoral vein catheter. Twenty minutes later arterial pressure registration was started and was continued for 40 minutes. The pulse-by-pulse values of systolic, diastolic and mean arterial pressure as well as the pulse intervals were measured by peak and rate detectors of the AcqKnowledge 2.0 software. Row data were processed using a virtual instrument developed in our laboratory in the graphical programming environment Lab VIEW 3.1.1. L-NAME increased systolic, diastolic and mean arterial pressure by 16.6%, 25% and 35%, respectively. The PMF/PHF ratio in heart rate spectrum decreased, indicating an increased vagal effect on the heart. Nitric oxide synthase inhibition increased the low-frequency component of systolic arterial blood pressure variability by 39.5%. Nitric oxide is a physiological regulator of rapid fluctuations of arterial blood pressure.

Atrial natriuretic peptide and dopamine in a dog model of acute renal ischemia.

Atrial natriuretic peptide (ANP) has been shown to reverse functional impairment in ischemic acute renal failure (ARF). To prolong and/or to enhance the effects of peptide, in this investigation dopamine (D) (3 micrograms/kg BW/min) was applied together with ANP (100 ng/kg BW/min) after 90 min unilateral renal artery occlusion in anesthetized dogs. ANP significantly increased creatine clearance, filtration fraction, diuresis, sodium excretion, sodium reabsorption, and free water clearance, as in postinfusion period only V remained elevated. D alone did not effect renal function beneficially. ANP+D improved kidney function impairment to a level comparable with that of ANP alone, but V and UNa.V remained increased in the postinfusion period. MAP was elevated during ANP+D infusion as compared to ANP alone and was sustained to the end of the experiment. We conclude that D does not potentiate the positive effects of ANP on postischemic kidney, but prolongs its action on UNa.V, possibly by maintenance of high MAP after renal ischemia.

Intact kidney function during contralateral renal artery clamping in dogs.

Experiments were carried out on 32 Nembutal anaesthetized mongrel dogs from both sexes. After 45 min control period unilateral renal ischemia was achieved by clamping the left renal artery for 90 min. In part of the experiments (n = 8) after clamp removal 3 consecutive 45 min periods were performed. The function of the intact right kidney was investigated. Mean arterial pressure (MAP), heart rate (HR), glomerular filtration rate (GFR), urine flow rate (V), fractional excretions of sodium (FENa), potassium (FEK) and chloride (FECl) and plasma levels of atrial natriuretic peptide, dopamine and antidiuretic hormone were evaluated. During ischemia MAP was elevated from 122.5 +/- 3.1 to 140.2 +/- 2.7 mmHg (p < 0.001), HR decreased from 119 +/- 4 to 102.5 +/- 3.9 beats/min (p < 0.01) as compared to the control period. GFR did not change significantly, while all excretory parameters increased: V from 8.7 +/- 1.2 to 14.5 +/- 1.7 microliters/min/gr kidney tissue (p < 0.05); FENa from 2.3 +/- 0.2 to 3.6 +/- 0.3% (p < 0.01); FEK from 40.0 < 3.5 to 51.2 < 2.8% (p < 0.05); FECl from 1.8 < 0.3 to 2.6 < 0.3% (p < 0.05). MAP remained elevated in the first and the second postischemic periods and was paralleled by the sustained increase in FENa and FECl, while FEK remained higher to the end of the experiment. ANP was significantly elevated during ischemia: on 75 min--p < 0.01 and on 105 min.--p < 0.05. AVP and dopamine showed no statistically significant changes during the investigated periods.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrial natriuretic peptide (ANP) as a neuropeptide: interaction with angiotensin II on volume control and renal sodium handling.

1. Angiotensin II (ANG II) and atrial natriuretic peptide (ANP) are functionally antagonistic circulating hormones involved in blood pressure and body fluid regulation. An inappropriate atrial secretion of ANP has been implicated in the pathogenesis of hypertension, but clinical and experimental results on the role of ANP in hypertension are still conflicting. 2. In the brain both peptides have been localized in close proximity, preferentially in areas involved in central cardiovascular, electrolyte and volume control. ANP was shown to inhibit ANG II-induced drinking, release of pituitary hormones and natriuresis, and to induce sodium retention when given alone. 3. These findings suggest that also in the brain ANG II and ANP exert functionally antagonistic effects. However, in contrast to their peripheral effects, ANG II induces natriuresis while ANP appears to cause antinatriuresis in the brain.

Excretory function after unilateral renal denervation and administration of propranolol to unanaesthetized dogs.

The experiments were carried out on female dogs with exteriorized ureters prior to and following surgical denervation of the left kidney. Propranolol 1.0 mg/kg b.w. was administered intravenously. Sodium, potassium, chloride, calcium, magnesium, zinc, copper, creatinine and urea concentrations in the urine from the denervated and intact kidneys as well as in blood drawn were determined. After renal denervation PAH clearance was determined. As a result of denervation diuresis and calcium and copper excretion were increased while urine osmolality was diminished. No change occurred in kidney blood flow and GFR. After propranolol administration diuresis, calcium and copper excretion in the intact kidney significantly increased. Changes in the excretory function of the left kidney following its denervation were not a result of alterations in renal haemodynamics. Results obtained indicative of that beta-adrenergic receptors contribute to the excretion of calcium and copper ions.

Excretory function of denervated kidney after inhibition of prostaglandin synthesis and furosemide administration in conscious dogs.

The experiments were carried out on unanaesthetized dogs with exteriorized ureters for separate urine collection from the left (denervated) and the right (intact) kidney. The osmolality and concentrations of sodium, potassium, calcium, magnesium, zinc, copper, chloride and creatinine were determined in the plasma as well as in the urine of the two kidneys. The function of the denervated and the innervated kidney was compared prior to and after indomethacin administration (5.0 mg/kg b.w.). The excretory function of both kidneys was also compared after furosemide treatment alone (0.5 mg/kg b.w.) as well as indomethacin pretreatment. Renal denervation increased urine flow rate, calcium and copper excretion. After administration, sodium excretion from the denervated kidney was higher than that from the intact one. Calcium excretion of the two kidneys did not differ significantly, while copper excretion from the denervated kidney was diminished, Furosemide administration after pretreatment with indomethacin did not lead to any difference between the denervated and intact kidney. The results show that renal nerves and prostaglandins participate jointly in the regulation of sodium, copper and calcium excretion. Renal prostaglandins do not change the response of the denervated kidney to furosemide as compared to the intact kidney.

Potentiation of the diuretic and natriuretic effect of furosemide by the calcium antagonist nifedipine.

The renal excretory function was studied in rats under control conditions, after treatment with nifedipine, with furosemide and combined treatment with nifedipine and furosemide. The experimental animals were investigated in metabolic cages. Spontaneously excreted urine was collected for 6 h (under control conditions and when nifedipine was applied) and for two hours in the experiments with furosemide treatment. The diuresis, the excretion of sodium, potassium, chlorine, creatinine and the total amount of osmotically active substances were studied. Blood samples were taken from the heart under nembutal anaesthesia. The clearances and the excretory fractions of the substances tested were determined. The glomerular filtration was determined by the clearance of the endogenous creatinine. The blood pressure in the tail artery was measured in control experiments. Increased diuresis was observed after treatment with nifedipine, without substantial changes in the sodium, chlorine, potassium and total osmotic excretion. When furosemide was applied after pretreatment with nifedipine, the diuresis was by 33 per cent higher than when only furosemide was applied (p less than 0.01), sodium excretion was by 40% higher (p less than 0.01) and that of chlorine--by 16.9% (p less than 0.01). Potassium excretion remained unchanged. The results of the excretory fractions and of the clearances of the substances studied support the evidence about potentiation of the natriuretic and chlorouretic effect of furosemide under the effect of nifedipine. The concentrating function of the kidney, determined through the clearance parameter Cosm/TcH2O manifested an additional limitation under the effect of nifedipine.

Excretory function of intact kidney after contralateral denervation on unanaesthetized dogs.

The experiments were carried out on unanaesthetized dogs with exteriorized ureters. The left kidney was denervated. The function of the left and right kidneys was compared prior to and after furosemide administration, 0.2 mg/kg b. w. The function of the right intact kidney prior to and after left kidney denervation was also compared. The concentrations of sodium, potassium, chloride, calcium, magnesium, zinc, copper, urea, and creatinine in plasma and urine were determined. Denervation of the left kidney was followed by an increase of diuresis, calcium, and copper excretion from the denervated kidney. After furosemide administration the chloride excretion from the side of denervation was also observed to increase. The changes in the excretory function of the left kidney after denervation were not accompanied by changes in excretory function of right (intact) kidney. The excretory function of the intact kidney after denervation of contralateral kidney and furosemide administration did not differ from that prior to denervation and furosemide administration. In unanaesthetized dogs no changes in the excretory function of intact kidney were observed as a result of contralateral kidney denervation.