Possible influence of intrarenal generation of kinins on prostaglandin release from the rabbit perfused kidney.

The effects of bradykinin and kininogen on renal prostaglandin release were studied in rabbit isolated kidneys perfused with oxygenated Krebs solution. The concentration of prostaglandin-like material in kidney effluent was determined by bioassay after extraction of the samples with organic solvents. In 7 experiments the samples were assayed after separation of prostaglandins E and F by thin layer chromatography. Addition of bradykinin to the perfusing fluid increased the venous and urinary effluxes of prostaglandin E-like substance by sixfold and fivefold, respectively, but efflux of prostaglandin F-like material was unaffected. Addition of kininogen to the perfusing fluid augmented the venous and urinary release of prostaglandin E-like substances by fifteenfold and ninefold respectively and caused a twofold increase in the efflux of prostaglandin F-like material into the venous effluent. Aprotinin, a kallikrein inhibitor, reduced the prostaglandin releasing action of kininogen but not of bradykinin. In contrast, inhibition of prostaglandin synthesis by indomethacin suppressed the release of prostaglandin evoked by either bradykinin or kininogen. This study suggests that augmented release of prostaglandins in response to kininogen is a consequence of renal generation of kinins. Thus, changes in the intrarenal activity of the kallikreinkinin system may modulate renal prostaglandin release.     

Effects of atrial natriuretic peptide on adrenergically induced norepinephrine release and vasoconstriction in the dog kidney.

The effects of atrial natriuretic peptide (ANP) on the neural control of renal blood flow were examined in anesthetized dogs. Intrarenal arterial infusion of ANP (alpha-hANP, 10 and 50 ng/kg per min) suppressed the decrease in renal blood flow but not the increase in renal venous plasma norepinephrine concentration induced by renal nerve stimulation (1 and 2 Hz, for 1 min). ANP also attenuated the blood flow response to intrarenal arterial injection of methoxamine (5-20 micrograms). These results suggest that ANP acts at a postsynaptic site to suppress adrenergically induced vasoconstriction in the dog kidney.     

Generation of prostaglandin E-like material by the guinea-pig trachea contracted by histamine.

Challenges of the superfused (1 ml min-1) trachea with histamine (100-200 mug) result in the release of prostaglandin E-like material (3-25 ng in terms of prostaglandin E2) but no prostaglandin F-like activity has been detected in the superfusate. This release is blocked by indomethacin (1mug ml-1) and then the contractile action of histamine is enhanced. It is concluded that the release of a prostaglandin E-like material by histamine from tracheal smooth muscles is a self-defensive mechanism protecting against the strong constriction of airways. The maximal relaxation of trachea by isoprenaline (50-500 mug) is not accompanied by the release of a prostaglandin-like material.     

Effects of adenosine on adrenergically induced renal vasoconstriction in dogs

The role of exogenous and endogenous adenosine in the neural control of renal blood flow was studied in anesthetized dogs. The plasma norepinephrine (NE) concentration was measured by high-performance liquid chromatography and the renal NE secretion rate was calculated. Renal nerve stimulation (1-3 Hz) reduced renal blood flow and increased NE secretion rate. The intrarenal arterial injection of NE (0.3-1.0 micrograms) also reduced renal blood flow. Infusion of adenosine (10-100 micrograms/min) into the renal artery attenuated the increase in NE secretion rate induced by renal nerve stimulation, but the nerve stimulation-induced decrease in renal blood flow was unaffected. On the other hand, adenosine potentiated the NE-induced renal blood flow response. Similar results were obtained with an adenosine potentiator, dipyridamole (1-10 micrograms/min). An adenosine receptor blocker, theophylline (0.3-1.0 mg/min), potentiated the NE secretion rate response induced by nerve stimulation, without any change in the renal blood flow response. The NE-induced renal blood flow response was attenuated by theophylline. These results suggest that adenosine inhibits neural NE release and enhances vasoconstriction in the dog kidney during sympathetic stimulation under in vivo conditions. These post- and presynaptic mechanisms may thus be activated by endogenous adenosine.     

Effects of endothelin on adrenergic neurotransmission in the dog kidney.

Endothelin infused into the renal artery (0.3-3 ng/kg per min) of anesthetized dogs suppressed the increases in renal venous plasma norepinephrine (NE) concentration and calculated NE efflux induced by renal nerve stimulation at low frequencies (0.7-1.2 Hz) but not at high frequencies (2-3.5 Hz). Endothelin failed to affect the decrease in renal blood flow induced by renal nerve stimulation or exogenous NE. These findings suggest that endothelin inhibits NE release during a slight rise in renal nerve activity, while it does not affect alpha-adrenoceptor-mediated vasoconstriction in the dog kidney.     

Graded effects of saralasin on prostaglandin E, plasma renin activity and renal blood flow in anaesthetized dogs

The objective of this study was to examine in anaesthetized dogs the graded effect of saralasin on renal prostaglandin E (PGE) release and to attempt to associate this change with its effects on plasma renin activity and renal blood flow. Blood pressure and renal blood flow were monitored. Renal PGE concentration and plasma renin activity were measured by radioimmunoassay. Saralasin or saline vehicle was infused into the renal artery for 20 min. Infusion of saralasin at the lowest dose of 0.25 micrograms/kg per min or saline vehicle did not alter either renal blood flow or plasma renin activity. Saralasin increased renal blood flow and caused a complete blockade of the renal vasoconstrictor response to exogenous angiotensin II at the two higher doses used (0.5 and 1 micrograms/kg per min). Only the highest dose of saralasin increased plasma renin activity significantly. Renal venous PGE concentration at the 5, 10 and 20 min periods of infusion was not changed significantly by any of these three doses of saralasin. We conclude therefore that the increases in renal blood flow and plasma renin activity caused by saralasin in the anaesthetized dog occur by mechanisms independent of changes in renal PGE.     

Effects of alpha,beta-methylene ATP on resistance and capacitance blood vessels of the cat intestinal circulation; a comparison with other vasoconstrictor agents and sympathetic nerve stimulation.

The autoperfused intestinal circulation of pentobarbitone anaesthetized cats was used to study the effects of alpha,beta-methylene ATP (1-100 micrograms i.a.) on pre-capillary resistance vessels and post-capillary capacitance (venous) blood vessels in comparison with other vasoconstrictor agents (also given i.a.) and the effects of sympathetic nerve stimulation (0.25-16 Hz). All cats were treated with atropine and propranolol. alpha,beta-Methylene ATP, noradrenaline and sympathetic nerve stimulation all caused dose- or frequency-dependent constriction of both resistance and capacitance vessels. alpha,beta-Methylene ATP was particularly active on capacitance vessels causing a greater constriction than either noradrenaline or sympathetic nerve stimulation. In comparison, angiotensin II and vasopressin caused a selective constriction of resistance vessels and prostaglandin F2 alpha a selective constriction of capacitance vessels. The results demonstrate that functional P2x purinoceptors are present on both arterial and venous blood vessels of the cat intestinal circulation.     

Effects of cocaine and nomifensine on canine renal venous norepinephrine and dopamine content during electrical stimulation of the renal nerves

Dopamine has been suggested to play a role in the regulation of renal blood flow following its neuronal release from within the kidney. Skepticism remains, however, as to whether a vascular dopaminergic innervation plays a physiologic role in renal blood flow regulation. Thus, to investigate this possibility, we examined whether electrical stimulation of the renal nerves evokes an overflow of dopamine into renal venous blood that could be augmented by cocaine or nomifensine, two inhibitors of presynaptic uptake. To verify uptake inhibition, the stimulation-induced overflow of norepinephrine (NE) was also examined. The increases in renal venous NE content observed during 0.5 and 2.0 Hz renal nerve stimulation were significantly increased by administration of cocaine (2.5 and 5.0 mg/kg) and nomifensine (1 mg/kg). Renal nerve stimulation (0.5 and 2.0 Hz) did not significantly increase free renal venous or total dopamine sulfate content before or following administration of either cocaine or nomifensine. Renal venous minus arterial dopamine content indicated that a neuronal release of dopamine into renal venous blood could not be clearly demonstrated in the canine kidney. These results do not support the existence of an active dopaminergic innervation of the renal vasculature involved in the regulation of renal blood flow.     

EFFECTS OF ZAPRINAST ON RENAL NERVE STIMULATION-INDUCED ANTI-NATRIURESIS IN ANAESTHETIZED DOGS

1. We examined whether zaprinast, a putative cGMP-specific phosphodiesterase inhibitor, affects neural control of renal function in pentobarbital-anaesthetized dogs. 2. Renal nerve stimulation (1Hz, 1ms duration) reduced urine flow rate, urinary Na⁺ excretion (UNaV) and fractional excretion of Na⁺ (FENa) with little change in either renal blood flow (RBF) or glomerular filtration rate (GFR). 3. Intrarenal arterial infusion of zaprinast (10 and 100 μg/kg per min) increased basal urine flow rate, UNaV and FENa but not RBF or GFR. Zaprinast infusion (100 μg/kg per min) also increased renal venous plasma cGMP concentration and urinary cGMP excretion. 4. Renal nerve stimulation-induced reductions in UNaV and FENa were attenuated during zaprinast infusion, whereas the reduction in urine flow rate was resistant to zaprinast. 5. Renal nerve stimulation increased the renal venous plasma noradrenaline concentration and renal noradrenaline efflux, which remained unaffected during infusion of zaprinast (100 μg/kg per min). 6. The results of the present study suggest that zaprinast induces natriuresis and counteracts adrenergically induced anti-natriuresis by acting on renal tubular sites in the dog kidney in vivo.     

Output of prostaglandins from the rabbit kidney, its increase on renal nerve stimulation and its inhibition by indomethacin

1. In each of six experiments, prostaglandins were identified in the renal venous blood of the rabbit. The concentrations in renal venous blood were up to 45 times higher than in aortic blood, suggesting that most of the prostaglandins originate from the kidney.2. Prostaglandins E(2), F(2a) and a prostaglandin of the A, C or B series were estimated by biological assay after solvent partition and column or thin-layer chromatography.3. Prostaglandins E(2) and F(2a) were identified conclusively by combined gas chromatography-mass spectrometry.4. Electrical stimulation of the renal nerve increased the output of prostaglandins.5. Indomethacin (10 mg/kg) injected intravenously, reduced the output of prostaglandins into renal venous blood and prevented the increase in output on renal nerve stimulation.     

Effect of SA-446, an angiotensin-converting enzyme inhibitor, on renal function in anesthetized dogs: special reference to arachidonic acid metabolites

Intravenous infusion of SA-446 (1 mg/min) decreased systemic blood pressure and increased renal blood flow in anesthetized dogs. These changes were accompanied by a slight natriuretic response. During the infusion of this dose of SA-446, the pressor response to angiotensin I was abolished and the depressor response to bradykinin was markedly potentiated. Administration of indomethacin (13 mg/kg i.v.) suppressed natriuresis and, to some extent, the renal vasodilation caused by SA-446. Before and after administration of SA-446, four arachidonate metabolites, prostaglandin E2, prostaglandin F2 alpha, 6-keto-prostaglandin F1 alpha, and thromboxane B2, were determined in plasma and urine by radioimmunoassay. There were no remarkable changes in levels of prostaglandins and thromboxane B2 in arterial and renal venous plasma. The urinary excretion of the metabolites varied little, but thromboxane B2 excretion did significantly decrease. Thus, reduction in the biosynthesis of thromboxane B2 in the kidney may relate to the effects of SA-446 on renal hemodynamics and urine formation.     

The effects of ionizing irradiation on production of thromboxane and prostacyclin by the isolated perfused rat kidney

Exposure to whole body radiation is associated with prompt changes in urinary excretion of prostaglandins. We investigated the separate effects of radiation on rat kidney capillary and tubular system prostaglandin synthesis. Animals were irradiated to the left kidney area with a single dose of 15 Gy. One week later the rats were anesthesized, the renal artery, vein and ureter of the left kidney cannulated, and the kidney removed and perfused with Krebs-Henseleit physiological buffer at a rate of 10-12 ml/min. Effluent fluids were collected separately from the renal vein and from the ureter of irradiated and control (operated sham-irradiated animals) and were assayed by radioimmunoassays for thromboxane A2 (TXB2) and prostacyclin (6 keto PGF1 alpha). Histological examination of the irradiated kidneys showed no significant changes, and electron microscopy revealed minimal interstitial edema. In contrast to these minimal changes, TXB2 assays showed a significant increase both in the venous and ureter effluents. Following stimulation with angiotensin II in the perfusate, a further significant increase in TXB2 production was observed both by the capillary and the tubular systems. With 6 Keto PGF1 a slightly different response was seen. The basal production was increased only in the ureter effluent of the irradiated animals, while there were no changes in the release in the venous effluents. In parallel, radiation significantly increased the angiotensin II stimulated production capacity of prostacyclin by the tubular system. The response of the capillary system following irradiation may create imbalance between these two important substances and lead to the radiation effects in the renal tissue.     

Inhibitory effect of cilazaprilat on norepinephrine release induced by renal nerve stimulation in anesthetized dogs

In pentobarbital-anesthetized dogs, the increase in the renal norepinephrine secretion rate elicited by renal nerve stimulation (1 Hz) during infusion of angiotensin I (15 ng/kg/min) was partially but significantly inhibited (by 21-37%) after dosing with cilazaprilat (0.1 mg/kg), an angiotensin converting enzyme inhibitor, accompanied by a decrease in the renal venous plasma norepinephrine concentration. These results may suggest that cilazaprilat exerts at least a part of its hypotensive effect through decreasing the facilitatory action of endogenous angiotensin II on adrenergic transmission.     

EFFECTS OF A NON-VASOCONSTRICTOR DOSE OF PHENYLEPHRINE ON PROSTAGLANDIN E2 AND RENIN RELEASE IN ANAESTHETIZED DOGS

Phenylephrine (1 μg/min) was infused into the renal artery of pentobarbitone anaesthetized dogs. Systemic blood pressure and renal blood flow were monitored. Arterial and renal venous blood samples were collected before and 10 min after the start of the infusion. Plasma prostaglandin E₂ concentration and plasma renin activity were measured by radioimmunoassay. This dose of phenylephrine did not affect systemic blood pressure and renal blood flow. However, both prostaglandin E₂ and renin secretion rates were increased by the infusion of phenylephrine. These results suggest that renal α-adrenoceptors participate in prostaglandin and renin release independent of the changes in systemic blood pressure and renal blood flow.     

EVIDENCE AGAINST THE INVOLVEMENT OF VASOACTIVE INTESTINAL PEPTIDE IN OVINE PAROTID SECRETION AND BLOOD FLOW

1. The proposition that stimulation of the secretomotor nerve to the ovine parotid gland might involve co-release of vasoactive intestinal peptide (VIP) was tested by studying responses to infusion of VIP directly into the gland's arterial blood supply and by assay of VIP in parotid venous blood. 2. In unstimulated glands, an arterial blood concentration of 1.5 - 2.5 X 10(-9) mol/L VIP did not evoke fluid secretion but it increased K+ and phosphate secretion and glandular blood flow. The same blood concentration of VIP potentiated the stimulation of salivary flow rate caused by intraarterial infusion of bethanechol but nerve stimulation was not potentiated. VIP increased glandular blood flow in both conditions of stimulation. 3. Atropine blocked neurally stimulated salivary secretion but an increase in glandular blood flow was still detectable. There was therefore no evidence for a non-cholinergic neural mechanism for salivary secretion. 4. Furthermore, VIP concentrations in glandular venous blood were not increased by nerve stimulation. 5. The results indicate that exogenous VIP can affect the flow and composition of ovine parotid secretion but was not involved in the response to secretomotor nerve stimulation.     

Renal function and renal venous prostaglandin concentrations during different stages of experimental renal hypertension in the rat.

Renal hypertension was produced in rats and the changes in renal function, renal venous prostaglandin E2 and F2alpha concentrations and secretion rates were studied at various times. Renal plasma flow transiently fell in the ischaemic kidney 2 weeks after clamping, whilst that of the other kidney did not change. Glomerular filtration rate remained constant in both kidneys throughout the entire study. Prostaglandins E2 and F2alpha concentrations rose in the venous plasma from the ischaemic kidney, but did not change in the other kidney and appeared to be inversely related to renal plasma flow. Calculated secretion rate of both prostaglandins fell in the ischaemic kidney and did not change in the other kidney. Clamping the second kidney, two weeks after the first, caused a further elevation in blood pressure, a fall in renal plasma flow and a fall in prostaglandin secretion rate in both kidneys. The implications of these prostaglandin changes are discussed.     

Interaction between norepinephrine release and intrarenal angiotensin II formation during renal nerve stimulation in dogs

We examined possible interactions between intrarenal angiotensin II (ANG II) formation and norepinephrine (NE) release during renal sympathetic nerve stimulation (RNS) in anesthetized dogs. During 10 min of continuous RNS (1.5-2 Hz), the ANG II formation rates (ANG II-FR) and NE secretion rates (NE-SR) were determined at 1 and 10 min. Under control conditions, almost the same extent of increase in the NE-SR was observed at 1 and 10 min of RNS, whereas a significant increase in ANG II-FR was observed at 10 min but not at 1 min. During intrarenal arterial infusion of enalaprilat or losartan, the increase in NE-SR and reduction in renal blood flow at 10 min of RNS were suppressed, whereas the NE release and vasoconstriction responses at 1 min remained unaffected. The RNS-induced increases in ANG II-FR were completely abolished during infusion of enalaprilat. These results suggest that NE release on continuous RNS is enhanced by concomitantly formed ANG II, and this interaction depends on the time-related changes in intrarenal ANG II formation during RNS in the canine kidney.     

Renal vascular and biochemical responses to systemic renin inhibition in dogs at low renal perfusion pressure.

Renin is produced by the kidney and secreted into the systemic circulation. However, its biochemical and physiological role of regulating renal blood flow with changing renal perfusion pressure (RPP) is not fully understood. In this study, the function of the intrarenal renin for production of angiotensin (Ang) I and maintenance of vascular tone was evaluated in dogs under normal conditions and when the kidney was perfused at low RPP. The dog left kidney was perfused first at normal (100 mm Hg) and then at low (30 mm Hg) RPP in the presence or absence of the renin inhibitor ciprokiren (3 mg/kg, i.v.). Both hemodynamic and biochemical parameters were measured. Lowering RPP markedly reduced left renal blood flow and elevated left renal vascular resistance. These effects were prevented by ciprokiren, which blocked the intrarenal production of Ang I. Lowering RPP increased the renal venous/ arterial ratio from 1.4+/-0.1 to 3.6+/-0.3 for plasma renin activity and from 2.4+/-0.2 to 9.8+/-1.1 for Ang I, but did not change the venous/arterial ratio for Ang II. The net renal venous conversion rate of Ang I to Ang II decreased from 0.22 to 0.09 after RPP was lowered, whereas the conversion rate in arterial blood was 1.35 and did not decrease significantly. Our results demonstrated the importance of intrarenal renin-angiotensin system for Ang I production and for the maintenance of the vascular tone, especially at low RPP. Our study also shows the limited capacity for Ang I conversion in the renal vasculature in vivo.     

PROSTAGLANDINS AND THE RENAL RESPONSES TO HAEMORRHAGE, ANGIOTENSIN II AND METHOXAMINE IN CONSCIOUS RABBITS

1. The responses to 20% haemorrhage were examined in conscious rabbits with or without inhibition of prostaglandin production by indomethacin (5 mg/kg +0.5 mg/kg per h i.v.). In rabbits not pretreated with indomethacin, haemorrhage lowered mean arterial pressure by 6.3 (s.e.m. = 1.6) mmHg, renal blood flow by 22.8 (s.e.m. = 3.4) ml/min and glomerular filtration rate (GFR) by 3.4 (s.e.m. = 0.6) ml/min, and raised plasma renin activity by 5.2 (s.e.m. = 1.0) ng/ml per h. Pretreatment of the rabbits with indomethacin did not significantly alter the responses to haemorrhage. Mean arterial pressure fell by 10.9 (s.e.m. = 1.8) mmHg, renal blood flow by 24.9 (s.e.m. = 3.9) ml/min and GFR by 4.2 (s.e.m. = 1.8) ml/min and plasma renin activity rose by 3.2 (s.e.m. = 0.5) ng/ml per h. 2. In a separate group of 5 rabbits, angiotensin II was infused at 10, 25 and 50 ng/kg per min i.v. or methoxamine was infused at 10 and 25 μg/kg per min i.v. After indomethacin pretreatment, angiotensin II caused a significantly greater rise in mean arterial pressure and greater fall in renal vascular conductance, but there was no effect on the GFR response. In contrast, methoxamine caused significantly smaller falls in GFR, renal blood flow and renal vascular conductance after indomethacin pretreatment. 3. Indomethacin significantly lowered resting GFR but not renal blood flow or arterial pressure. 4. Thus, indomethacin pretreatment accentuated the renal vasoconstriction to angiotensin II, reduced the renal vasoconstriction to methoxamine and had no effect on the responses to haemorrhage. 5. The results therefore suggest that prostaglandins do not act to lessen the renal effects of all vasoconstrictor stimuli, but that the prostaglandin response depends on the nature of the ischaemic stimulus.     

Renal hemodynamic responses to intrarenal infusion of ligands for the putative angiotensin IV receptor in anesthetized rats.

Angiotensin IV, a hexapeptide fragment (3-8) of angiotensin II metabolism, has been reported to produce vasodilatation within the renal vasculature by activation of the putative AT4 receptor. However, there are conflicting findings, with previous in vivo studies providing evidence for and against a renal vasodilator action of angiotensin IV. In this study, the renal hemodynamic responses to activation of the putative AT4 receptor were studied in anesthetized rats by left renal arterial infusion of two endogenous ligands, angiotensin IV and LVV-hemorphin-7. Angiotensin IV (10, 100, and 1,000 pmol/min) infusion caused dose-dependent reductions in blood flow to the infused kidney, which were abolished by pretreatment with losartan. In respect to this effect, angiotensin IV was approximately 300-fold less potent than angiotensin II. There were no significant effects of angiotensin IV on mean arterial pressure, heart rate, or blood flow to the noninfused kidney. Intrarenal infusion of LVV-hemorphin-7 (10, 100, and 1,000 pmol/min) had no significant effect on renal blood flow in the infused and noninfused kidneys, or on mean arterial pressure or heart rate. These results provide no evidence for a renal vasodilatory action of angiotensin IV or LVV-hemorphin-7. On the contrary, intrarenal angiotensin IV infusion produced vasoconstriction of the renal vasculature, mediated by activation of AT1 receptors. These observations provide evidence against a vasodilatory role of putative AT4 receptors in the rat kidney.