High salt intake increases blood pressure in normal rats: putative role of 20-HETE and no evidence on changes in renal vascular reactivity.

UNLABELLED: Background/Aims . High salt (HS) intake may elevate blood pressure (BP), also in animals without genetic salt sensitivity. The development of salt-dependent hypertension could be mediated by endogenous vasoactive agents; here we examined the role of vasodilator epoxyeicosatrienoic acids (EETs) and vasoconstrictor 20-hydroxyeicosatetraenoic acid (20-HETE). METHODS: In conscious Wistar rats on HS diet systolic BP (SBP) was examined after chronic elevation of EETs using 4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (c-AUCB), a blocker of soluble epoxide hydrolase, or after inhibition of 20-HETE with 1-aminobenzotriazole (ABT). Thereafter, in acute experiments the responses of renal artery blood flow (Transonic probe) and renal regional perfusion (laser-Doppler) to intrarenal acetylcholine (ACh) or norepinephrine were determined. RESULTS: HS diet increased urinary 20-HETE excretion. The SBP increase was not reduced by c-AUCB but prevented by ABT until day 5 of HS exposure. Renal vasomotor responses to ACh or norepinephrine were similar on standard and HS diet. ABT but not c-AUCB abolished the responses to ACh. Conclusions . 20-HETE seems to mediate the early-phase HS diet-induced BP increase while EETs are not engaged in the process. Since HS exposure did not alter renal vasodilator responses to Ach, endothelial dysfunction is not a critical factor in the mechanism of salt-induced blood pressure elevation.

Treatment efficacy of hypertension in kidney transplant recipients in the Netherlands.

BACKGROUND: Hypertension in kidney transplant recipients jeopardises graft and patient survival. Guidelines suggest blood pressure targets of ≤130/80 mmHg and sodium intake <90 mmol/day. METHODS: Since the efficacy of antihypertensive treatment among kidney transplant recipients is unknown, we analysed data on office-based blood pressure and use of antihypertensive drugs from the Netherlands Organ Transplant Registry on 5415 kidney transplant recipients. Additionally, we studied dosages, prevalence of treatment-resistant hypertension and 24-hour sodium excretion in 534 kidney transplant recipients from our centre to explore possibilities for therapy optimisation. RESULTS: In patients registered in the Netherlands Organ Transplant Registry, median blood pressure was 134/80 mmHg (interquartile range 122-145/70-85). In 77.2%, the blood pressure was ≥130/80 mmHg; of these patients 10.4% had no registered use, 30.0% used one and 25.9% used ≥3 classes of antihypertensive agents. Parameters from our centre were comparable: 78.7% had a median blood pressure of ≥130/80 mmHg of whom 14.5% had no registered use of antihypertensives and 26.4% used ≥3 classes. Sub-maximal dosages were prescribed in 74.0% of the kidney transplant recipients with a blood pressure of ≥130/80 mmHg while using at least one antihypertensive agent. Treatment-resistant hypertension was present in 7.7%. Median 24-hour sodium excretion was 147 mmol/day (interquartile range 109-195). CONCLUSIONS: This study suggests that therapeutic optimisation of antihypertensive treatment in kidney transplant recipients is, in theory, frequently possible by intensifying pharmacological treatment and by providing more advice on dietary sodium restrictions.

Assessment of small bowel function in critical illness: potential role of citrulline metabolism.

Small intestinal function in critically ill patients should ideally be assessed in order to determine the preferred feeding route, timing, and composition of enteral nutrition. Additionally, evaluation of small bowel function may lead to new insights aimed to maintain enterocyte integrity. Critically ill patients are likely to have impaired enterocyte function mainly as a consequence of diminished splanchnic blood flow associated with mucosal hyperpermeability and bacterial translocation, a pathological state believed to be pivotal in the development of sepsis and multiple organ dysfunction syndrome (MODS). However, feasible and validated clinical tools to reliably assess enterocyte function are lacking. This explorative review discusses the promising role of citrulline, a nonprotein amino acid almost exclusively generated by the enterocyte, as a biomarker reflecting enterocyte function in critically ill patients. Citrulline metabolism, its potential as enterocyte biomarker, and literature on citrulline in critically illness will be discussed. Finally, a novel test for enterocyte function, the citrulline generation test (enterocytic citrulline production upon stimulation with enteral or intravenous glutamine) will be considered briefly.

Effects of ATP on rat renal haemodynamics and excretion: role of sodium intake, nitric oxide and cytochrome P450.

AIM: Adenosine-5'-triphosphate (ATP) affects intrarenal vascular tone and tubular transport via P2 receptors; however, the actual role of the system in regulation of renal perfusion and excretion remains unclear and is the subject of this whole-kidney study. METHODS: Effects of suprarenal aortic ATP infusion, 0.6-1.2 mg kg(-1) h(-1), were examined in anaesthetised rats maintained on low- (LS) or high-sodium (HS) diet. Renal artery blood flow (RBF, transonic flow probe) and the perfusion (laser-Doppler flux) of the superficial cortex (CBF) and outer and inner medulla (OM-BF, IM-BF) were measured, together with sodium and water excretion and urine osmolality. RESULTS: Adenosine-5'-triphosphate did not change arterial pressure, RBF or CBF while the effects on medullary perfusion depended on sodium intake. In LS rats ATP increased IM-BF 19 +/- 6%, the effect was prevented by inhibition of nitric oxide (NO) with N-nitro-l-arginine methyl ester. In HS rats ATP decreased OM-BF 16 +/- 3% and IM-BF (7 +/- 4%, not significant); previous inhibition of cytochrome P450 with 1-aminobenzotriazol blunted the OM-BF decrease and reversed the previous decrease of IM-BF to a 13 +/- 8% increase. Inhibition of P2 receptors with pyridoxal derivative (PPADS) abolished medullary vascular responses to ATP. In HS rats pre-treated with PPADS, ATP increased tubular reabsorption, probably via adenosine formation and stimulation of P1 receptors. CONCLUSION: The data indicate a potential role of ATP in the selective control of renal medullary perfusion, different in sodium depleted and sodium replete rats. The action of ATP appears to be mediated by the NO system and the cytochrome P450 dependent vasoactive metabolites.

Renal vascular effects of frusemide in the rat: influence of salt loading and the role of angiotensin II.

We showed recently that post-frusemide (furosemide) natriuresis was associated with a major depression of medullary circulation. In the present study, prior to administration of frusemide the tubular transport of NaCl was modified by loading the animals with 5% saline to elucidate a possible interrelation between the tubular and vascular effects of the drug. Moreover, a possible involvement of the renin-angiotensin system was examined by pharmacological blockade using captopril, an inhibitor of angiotensin converting enzyme (1 mg x kg(-1), I.V.), or losartan, a selective inhibitor of angiotensin AT1 receptor (10 mg x kg(-1), I.V.). The effects of frusemide (0.25 mg x kg(-1) I.V., then the same dose given over 1 h) on renal medullary and cortical circulation (using laser-Doppler flowmetry) and renal excretion of sodium (U(Na)V), water and total solutes were measured in anaesthetised rats. With no pre-treatment, frusemide decreased the medullary flow (36.6 +/- 6.0%) significantly more than the cortical flow (10.1 +/- 1.0%; P < 0.001). The difference between the medulla and cortex was not significant in rats which showed high U(Na)V after hypertonic saline loading (2.0 +/- 0.4 vs. 0.4 +/- 0.1 micromol x min(-1) in non-loaded rats): 21.1 +/- 3.9% and 15.8 +/- 1.5%, respectively. At very high U(Na)V (9.5 +/- 1.1 micromol x min(-1)) the post-frusemide decrease in blood flow tended to be smaller in the medulla (7.6 +/- 7.7%) than in the cortex (16.2 +/- 2.6%). The fall in medullary blood flow was attenuated by pre-treatment with captopril (22.0 +/- 3.3%) and abolished by pre-treatment with losartan (2.8 +/- 11.8%). The decrease in cortical blood flow was not changed by hypertonic saline or angiotensin II blockers. The abolition of the post-frusemide depression of medullary blood flow by previous salt loading confirms the proposed link between tubular transport status and vasoconstriction. A similar modification of the response by blockade of the renin-angiotensin system suggests that the system is involved in the mechanism of medullary vasoconstriction.

Differential effect of frusemide on renal medullary and cortical blood flow in the anaesthetised rat.

In addition to its known effect on renal tubular transport, frusemide (furosemide) has been shown to affect renal circulation. This study in the anaesthetised rat examined the influence of frusemide (bolus 0.25 or 0.5 mg kg(-1) I.V., then infusion delivering the same dose over 1 h) on renal cortical and medullary circulation measured as laser-Doppler blood (cell) flux. The responses were compared with simultaneously measured changes in renal excretion and in the tissue admittance, an index of medullary ionic hypertonicity of the interstitium. Renal vascular responses to frusemide were significant but not dose dependent. During low-dose frusemide infusion cortical flux decreased 11.5 +/- 0.9% and medullary flux decreased 32.3 +/- 3.5% (difference significant at P < 0.001). During high-dose infusion the decreases were by 13.5 +/- 1.4 and 29.3 +/- 3.8%, respectively (difference significant at P < 0.001). Sodium excretion increased 15-fold (by 3.7 +/- 0.4 micromol x min(-1)) and 30-fold (by 5.9 +/- 1.1 micromol x min(-1)) during low- and high-rate infusion of frusemide, respectively. By contrast, medullary tissue admittance decreased similarly with the two doses: maximally by 13.4 +/- 1.4 and 10.9 +/- 0.9%, respectively. The observations that an exaggerated post-frusemide decrease in blood flow within the medulla coincided with decreasing tissue admittance in this zone and that neither medullary blood flow nor admittance changes were related to the dose suggest a causal relationship between interstitial ionic hypertonicity and vascular resistance. We propose that the post-frusemide decrease in medullary tissue NaCl depressed medullary circulation by inhibiting local generation of vasodilator prostaglandins.

Osmotic hypertonicity of the renal medulla during changes in renal perfusion pressure in the rat.

1. The relationship between renal perfusion pressure (RPP) and ion concentration in renal medulla was studied in anaesthetized rats. RPP was changed in steps within the pressure range 130-80 mmHg, while tissue electrical admittance (Y, index of interstitial ion concentration) and medullary and cortical blood flow (MBF and CBF; laser Doppler flowmetry) were measured, along with glomerular filtration rate (C in) and renal excretion. 2. With a RPP reduction from 130 to 120 mmHg, tissue Y remained stable; at 100 and 80 mmHg, Y was 5 and 17 % lower, respectively, than at 120 mmHg. 3. CBF fell less than RPP (partial autoregulation) in the range 130-100 mmHg only. MBF was autoregulated within 120-100 mmHg, but not above or below this range. 4. Each step of RPP reduction was followed by a decrease in sodium and water excretion (UNaV and V). The osmolality of excised inner medulla fragments was similar at 120 and 105 mmHg (586 +/- 45 and 618 +/- 35 mosmol (kg H2O)-1, respectively) but lower at 80 mmHg (434 +/- 31 mosmol (kg H2O)-1, P < 0.01); the ion concentration changed in parallel. 5. The data show that medullary hypertonicity was well preserved during RPP fluctuations within 130-100 mmHg, but not below this range. RPP-dependent changes of UNaV and V were not clearly associated with changes in solute concentration in medullary tissue.

Simultaneous recording of tissue ion content and blood flow in rat renal medulla: evidence on interdependence.

The relationship of renal medullary tissue ion concentration and medullary blood flow (MBF) has never been closely evaluated because of limitations of available measuring methods. In an attempt to overcome this difficulty, an integrated probe was developed for simultaneous recording in rat renal medulla of tissue electrical admittance (Y), an index of interstitial ion concentration, and tissue perfusion with blood (laser-Doppler method). During spontaneous-selective MBF variations tissue Y showed inverse changes (r = -0.77, P < 0.001). The inverse correlation of the two variables was also seen after MBF has been reduced (-43%) by indomethacin, 5 mg/kg body wt iv (r = -0.77, P < 0.01). A modest selective MBF reduction (15%) induced by glibenclamide, an inhibitor of ATP-dependent K channels, did not alter medullary tissue admittance. The data support experimentally the concept that the rate of medullary tissue perfusion with blood is one determinant of interstitial solute concentration; however, changes in the latter were demonstrable only with major alterations of the MBF.

Cardiovascular and renal effects of endothelin receptor blockade with PD 145065 and interaction with urodilatin.

Role of endogenous endothelins (ET) in the control of cardiovascular system and renal function, and ET interaction with urodilatin (URO) were studied in anaesthetized rats. Activity of ET was blocked using PD 145065 (5 mg kg-1 body wt i.v.) a non-selective antagonist of ETA and ETB receptors. PD 145065 decreased mean arterial blood pressure (MBP) from 114 +/- 4 to 109 +/- 4 mmHg and the renal blood flow (RBF) from 6.6 +/- 0.3 to 5.8 +/- 0.4 mL min-1 (P < 0.02) and increased renal vascular resistance (RVR) from 17.7 +/- 1.2 to 20.1 +/- 2.1 mmHg min mL-1. Heart rate (HR) and renal function were not affected. URO was infused i.v. at 0.1 nmol min-1 kg-1 body wt without or with previous ET receptor blockade. After pre-treatment with PD 145065, infusion of URO decreased MBP more than did URO alone: 15 +/- 3% vs. 7 +/- 2% (P < 0.05). RVR and HR did not change after URO alone but decreased with URO given to PD 145065 treated rats (19 +/- 5% and 14 +/- 3%, respectively, P < 0.01). It is concluded that in anaesthetized surgically prepared rats endogenous ET can cause renal vasodilation, in contrast to constriction of systemic vasculature. Enhancement by ET blockade of vascular systemic and renal effects of URO supports ET interaction with natriuretic peptides in the control of cardiovascular and renal function.

Nitric oxide modulates angiotensin II- and norepinephrine-dependent vasoconstriction in rat kidney.

This study compared the vasoconstrictor action of angiotensin II (ANG II) and norepinephrine (NE) with different levels of nitric oxide (NO) in the kidney of anesthetized rats. In one series of experiments, the drugs were infused intravenously, and systemic NO content was reduced by a NO synthase inhibitor, nitro-L-arginine methyl ester (L-NAME). L-NAME significantly enhanced the renal blood flow (RBF) reduction produced by ANG II from 26 to 49%, but it had no significant effect on the change in RBF induced by NE. Medullary blood flow was not influenced by either ANG II or NE given alone or given after L-NAME. In the second series of experiments, all drugs were infused into the renal artery to avoid their systemic and, hence, extrarenal effects. In these experiments, renal content of NO was increased by the NO donor sodium nitroprusside (SNP), decreased by L-NAME, or restored by replacing endogenous NO by exogenous NO (L-NAME + SNP). Effects of both ANG II and NE on RBF were similarly and significantly attenuated by SNP (60% of control), enhanced by L-NAME (200% of control), and restored by L-NAME + SNP (90% of control, not significant). Our results indicate that NO attenuates the renal vasoconstriction due to ANG II or NE and that the antagonism between vasoconstrictors and NO is not due to a constrictor-induced production of NO because exogenous and endogenous NO were equally effective.

Modulation of renal medullary ionic hypertonicity by prostaglandins: data from tissue admittance studies in the rat.

1. Modulation of the cortico-papillary electrolyte gradient by prostaglandins (PG) was studied in the kidney of anaesthetized rats. The intrarenal PG activity was varied by synthesis blockade with indomethacin (Ind) or meclophenamate (Me) and by intrarenal infusion of prostaglandin E2 (PGE2). 2. The interstitial electrolyte concentration in the medulla was continuously recorded in the kidney in situ as tissue electrical admittance (reciprocal impedance); the total renal blood flow (RBF), inulin clearance (Cin) and renal excretion were measured simultaneously. 3. Indomethacin and Me (15 mg kg-1 h-1) increased tissue admittance 15-20% in the inner and 12-15% in the outer medulla (P < 0.001) whereas PGE2 (300 ng kg-1 min-1) decreased admittance 14 and 8%, respectively (P < 0.01). 4. Renal blood flow and Cin were not affected by intrarenal PG activity changes. There was an increase in urine concentration after PG blockade and a delayed decrease after PGE2 infusion. 5. A joint analysis of the dynamics of medullary tissue admittance, renal haemodynamics and renal excretion provides evidence that PGs modify the medullary ionic hypertonicity by affecting NaCl transport in the ascending limb of the loop of Henle.

Role of vasopressin V2 receptors in modulation of the renal cortico-papillary NaCl gradient.

In order to examine if modulation by vasopressin of NaCl transport in Henle's loops (via V2 receptors) can significantly modify medullary ionic hypertonicity, the effects of stimulation or inhibition of these receptors were studied in anaesthetized Wistar rats. Total electrolyte concentration in the medullary interstitium was continuously measured as tissue admittance (reciprocal impedance), using needle electrodes recording from the inner and outer medulla of the in situ kidney. Deamino-[Cys1, D-Arg8]vasopressin (dDAVP], a V2 agonist, infused i.v. at 7.5 ng.min-1.kg-1, significantly increased admittance by 9% and 8% in the inner and outer medulla, respectively. A slightly pressor i.v. infusion of natural arginine vasopressin (AVP) induced pressure natriuresis and did not affect medullary electrolyte concentration. Inhibition of V2 receptors with [d(CH2)5, D-Phe2, Ile4]-AVP, infused i.v. at 133 micrograms.h-1 kg-1 in indomethacin-treated rats, decreased admittance (significant in the inner medulla). Neither of the three agents used caused significant changes in the renal blood flow (RBF) or clearance of inulin (Cin). The demonstration that changing activity of V2 receptors affects the corticopapillary NaCl gradient indicates that, at least in rodents, stimulation of loop salt transport by AVP may represent an additional mechanism enhancing urine concentration.

Conductance studies of rat renal medulla for rapid estimation of extracellular electrolyte concentration.

A method is described for estimation of tissue extracellular electrolytes in fresh excised rat renal medulla on the basis of its electrical conductance. The inner medulla was placed between two standard flat electrodes and the conductance was measured at a frequency of 3.5 kHz. The data were corrected for the surface area and thickness of the tissue fragment. The standard conductance so determined clearly correlated with total electrolyte concentration, and Na+ and 2(Na+ + K+) concentrations in the whole tissue (r = 0.84, 0.81 and 0.77, respectively; P < 0.001). The method enables a rapid and simple estimation of medullary extracellular electrolytes, a variable that is physiologically more meaningful than whole tissue electrolyte concentration.

Cysteine enhances in vivo natriuretic potency of ethacrynic acid in the rat.

Total renal blood flow, glomerular filtration rate, and renal excretory function were determined in anesthetized rats treated with intravenous infusion of ethacrynic acid, 0.36 mg.min-1.kg-1, alone or in combination with cysteine. Simultaneously, the corticomedullary electrolyte gradient was evaluated in vivo from measurement of tissue electrical admittance (reciprocal impedance). Renal hemodynamics was not altered by drug infusion. Sodium excretion increased 1.7-fold with ethacrynic acid alone and 5-fold after the addition of cysteine. Tissue electrolytes of inner medulla decreased much more in rats given ethacrynic acid plus cysteine. We conclude that the addition of cysteine to intravenous infusion of ethacrynic acid greatly enhances its in vivo natriuretic potency in the rat.

Atrial peptide natriuresis in the rat without genuine rise in filtration rate or wash-out of medullary electrolytes.

1. Effects of synthetic atrial natriuretic peptide (ANP) on renal excretion, total renal blood flow (RBF), glomerular filtration rate (GFR) and tissue electrical admittance (reciprocal impedance, an estimate of tissue electrolytes) were determined in pentobarbitone-anaesthetized rats. GFR was measured both as inulin clearance (Cin) and as a product of renal plasma flow (RPF) and inulin extraction ratio (Ein). 2. With the lowest dose of ANP (0.35 micrograms/(kg min) I.V.) a 5-fold increase in sodium excretion occurred without measurable change in Cin, RPF x Ein nor medullary electrolyte concentration estimated from tissue electrical admittance. 3. With medium and high dosage (2 and 6 micrograms/(kg min), respectively), major and rapid increases in sodium excretion and urine flow were associated with an acute increase in Cin but not RPF x Ein. 4. The RBF increase observed in all groups of rats was not dose-related and did not parallel the natriuresis. Electrolyte concentration in the medullary tissue showed a modest transient decrease in rats given medium and high ANP doses. 5. We conclude that pronounced ANP natriuresis can develop in the absence of a measurable increase of GFR, estimated by a method not subject to urinary dead space error (RPF x Ein). The small transient decrease in medullary tissue electrolytes observed with higher peptide doses does not support solute wash-out as an important mechanism of increased sodium excretion.