Intrarenal Nitric Oxide Production Research Articles

Regulatory and functional interaction of vasoactive factors in the kidney and extracellular pH

A growing body of evidence suggests that vasoactive factors produced in the kidney such as nitric oxide, endothelins, angiotensin, and prostaglandins participate actively in the regulation of acid-base homeostasis under physiologic conditions. In addition, recent reports indicate that alterations in the systemic acid-base status may also influence the generation of vasoactive cytokines in the kidney, which in turn may mediate the renal effector processes that tend to restore normality under such conditions. Metabolic acidosis, which so frequently accompanies many forms of chronic renal failure (CRF), may contribute to down-regulation of intrarenal nitric oxide production that characterizes CRF. Reduced extracellular pH inhibits inducible nitric oxide production in mesangial cells by altering the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidation, an important posttranslational mechanism in the inducible nitric oxide synthase (iNOS) activation. The underlying defects resulting in the uncoupling of NADPH oxidation in acidemic microenvironment are discussed. Acidosis stimulates renal production of endothelins, which mediate proximal tubular acidification by enhancing sodium-hydrogen exchanger-3 (NHE-3) activity. Renal endothelins mediate enhanced urinary acid excretion following dietary acid ingestion, an effect that is effectively blocked by endothelin receptor blockers. Reduced extracellular pH stimulates endothelin secretion from renal microvascular endothelial cells, which may promote enhanced acid excretion from the distal tubule under conditions of acidosis. These phenomena as well as the role of angiotensin and renal prostaglandins in mediating renal acidification in normal and acidotic conditions are discussed in this review, which describe the regulatory interaction between extracellular pH and renal vasoactive factors.

Role of nitric oxide in diabetic nephropathy

Diabetic nephropathy is the leading cause of end-stage renal disease in the Western hemisphere. Endothelial dysfunction is the central pathophysiologic denominator for all cardiovascular complications of diabetes including nephropathy. Abnormalities of nitric oxide (NO) production modulate renal structure and function in diabetes but, despite the vast literature, major gaps exist in our understanding in this field because the published studies mostly are confusing and contradictory. In this review, we attempt to review the existing literature, discuss the controversies, and reach some general conclusions as to the role of NO production in the diabetic kidney. The complex metabolic milieu in diabetes triggers several pathophysiologic mechanisms that simultaneously stimulate and suppress NO production. The net effect on renal NO production depends on the mechanisms that prevail in a given stage of the disease. Based on the current evidence, it is reasonable to conclude that early nephropathy in diabetes is associated with increased intrarenal NO production mediated primarily by constitutively released NO (endothelial nitric oxide synthase [eNOS] and neuronal nitric oxide synthase [nNOS]). The enhanced NO production may contribute to hyperfiltration and microalbuminuria that characterizes early diabetic nephropathy. On the other hand, a majority of the studies indicate that advanced nephropathy leading to severe proteinuria, declining renal function, and hypertension is associated with a state of progressive NO deficiency. Several factors including hyperglycemia, advanced glycosylation end products, increased oxidant stress, as well as activation of protein kinase C and transforming growth factor (TGF)-β contribute to decreased NO production and/or availability. These effects are mediated through multiple mechanisms such as glucose quenching, and inhibition and/or posttranslational modification of NOS activity of both endothelial and inducible isoforms. Finally, genetic polymorphisms of the NOS enzyme also may play a role in the NO abnormalities that contribute to the development and progression of diabetic nephropathy.

Acute Erythropoietin Infusion Increases Rat Glomerular Filtration Rate by Partly Stimulating Intrarenal Nitric Oxide Production

Currently, erythropoietin (EPO) treatment in anemic patients is getting more interest, but in long term, development of hypertension is common problem in EPO treated patients. Since it isn't yet known whether EPO affects directly vascular and/or kidney functions, we aimed to study effects of acute EPO treatment on renal tissue nitrite level, and cardiovascular function and glomerular filtration rate (GFR). Experiments were done under 4 groups. Group I: Sham operated control, group II: 150 IU/kg EPO, group III: 50 mg/kg Nw-nitro-L-arginine methyl ester (L-NAME), group IV: L-NAME+EPO. Femoral artery, vein and bladder were catheterized under anesthesia. After stabilization and first 45 min basal control period, drugs were given i.v. bolus infusion according to the group protocols, then 2 × 45 min clearance periods were followed. EPO infusion increased renal tissue nitrite activity causing GFR increase without any influence on systemic blood pressure. L-NAME alone or together with EPO significantly raised the systemic blood pressure with a partial increase in GFR, but L-NAME treatment significantly reduced tissue nitrite level. The present study for the first time suggests that exogenous EPO treatment increases rat GFR and renal tissue nitrite level without affecting systemic blood pressure.

DOI, a 5-HT 2 receptor agonist, induces renal vasodilation via nitric oxide in anesthetized dogs

We have previously reported that (±)-1-(2.5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), a 5-HT 2 receptor agonist, induced renal vasodilation in anesthetized dogs. The present study was designed to investigate whether DOI-induced renal vasodilation might be mediated by increased nitric oxide (NO) release/production in renal tissue. The experiments were performed in anesthetized dogs. A 23-gauge needle was inserted into the left renal artery for infusion of drug solutions. Renal blood flow was measured with an electromagnetic flowmeter. The microdialysis probes were implanted into the renal cortex to collect the dialysate for measurement of guanosine 3′,5′-cyclic monophosphate (cGMP) and nitrite/nitrate (NO 2/NO 3) concentration. Intrarenal infusion of DOI at a rate of 5 μg/kg/min resulted in a significant increase, by 30±4%, in renal blood flow, indicating renal vasodilation. The renal interstitial concentrations of NO 2/NO 3 and cGMP also increased by 70±6% and 60±6%, respectively. These changes induced by DOI were completely abolished by the intrarenal pretreatment with N w-nitro- l-arginine methyl ester ( l-NAME, a NO synthase inhibitor, 100 μg/kg/min) or sarpogrelate (100 μg/kg/min, a highly selective 5-HT 2 receptor antagonist). DOI infusion increased urine volume and urinary excretion of Na +, which were also blocked by l-NAME or sarpogrelate. These results suggest that DOI caused renal vasodilation due to increased NO release/production by stimulation of 5-HT 2 receptors in the kidney. The natriuretic effect of DOI might also be related to increased intrarenal NO production.

Nitric oxide in the control of renal hemodynamics and excretory function

Experimental evidence has now been amassed to indicate that inhibition of nitric oxide (NO) synthase reduces total or regional renal blood flow by approximately 25 to 30% and markedly increases the renal vascular resistance, demonstrating that basal release of NO helps to maintain the relatively low vascular resistance that is characteristic for the kidney. It has been demonstrated that intraarterial administration of NO synthase inhibitors causes marked reductions in sodium excretion without changes in filtered load and suppressed the arterial pressure-induced natriuretic responses in the kidney. We also demonstrated that a constant rate infusion of a NO donor in dogs pretreated with a NOS inhibitor resulted in increases in sodium excretion but failed to restore the slope of the relation between arterial pressure and sodium excretion, suggesting that an alteration in intrarenal NO production rate during changes in arterial pressure is involved in the mediation of pressure natriuresis. Further experiments in dogs performed in our laboratory have confirmed that there is a direct relationship between changes in arterial pressure and intrarenal NO activity measured using NO-sensitive microelectrodes in the renal tissue. These arterial pressure-induced changes in intrarenal NO activity were seen positively correlated with the changes in urinary excretion rates of sodium. Collectively, these data suggest that acute changes in arterial pressure alter intrarenal NO production, which inhibits tubular sodium reabsorption to manifest the phenomenon of pressure natriuresis.

Hyperuricemia exacerbates chronic cyclosporine nephropathy.

Hyperuricemia frequently complicates cyclosporine (CSA) therapy. The observation that longstanding hyperuricemia is associated with chronic tubulointerstitial disease and intrarenal vasoconstriction raised the hypothesis that hyperuricemia might contribute to chronic CSA nephropathy. CSA nephropathy was induced by the administration of CSA (15 mg/kg/day) for 5 and 7 weeks to rats on a low salt diet (CSA group). The effect of hyperuricemia on CSA nephropathy was determined by blocking the hepatic enzyme uricase with oxonic acid (CSA-OA). Control groups included rats treated with vehicle (VEH) and oxonic acid alone (OA). Histological and functional studies were determined at sacrifice. CSA treated rats developed mild hyperuricemia with arteriolar hyalinosis, tubular injury and striped interstitial fibrosis. CSA-OA treated rats had higher uric acid levels in association with more severe arteriolar hyalinosis and tubulointerstitial damage. Intrarenal urate crystal deposition was absent in all groups. Both CSA and CSA-OA treated rats had increased renin and decreased NOS1 and NOS3 in their kidneys, and these changes are more evident in CSA-OA treated rats. An increase in uric acid exacerbates CSA nephropathy in the rat. The mechanism does not involve intrarenal uric acid crystal deposition and appears to involve activation of the renin angiotensin system and inhibition of intrarenal nitric oxide production.

Furosemide prevents the inhibitory effect of cyclosporine on intrarenal nitric oxide production

Furosemide prevents the inhibitory effect of cyclosporine on intrarenal nitric oxide production

University of Wisconsin preservation solution enhances intrarenal nitric oxide production

University of Wisconsin preservation solution enhances intrarenal nitric oxide production

Angiotensin II and its receptors in the diabetic kidney

Studies using either angiotensin-converting enzyme inhibitors or type 1 (AT1 ) angiotensin II (ANG II)-receptor blockers indicate that ANG II is a mediator of progressive injury in diabetic nephropathy. However, suppression of the systemic renin-angiotensin system (RAS) generally has been shown in diabetes mellitus. Evidence suggests that intrarenal RASs within glomeruli and proximal tubules may be activated with hyperglycemia, leading to stimulation of local ANG II production, which may exert feedback inhibition of systemic renin release. Once formed, intrarenal ANG II exerts most of its well-described effects through binding to AT1 receptors that are abundantly present in cells of the glomeruli, tubules, vasculature, and interstitium. Thus, AT1 -receptor activation increases vascular resistance, reduces renal blood flow, and stimulates production of extracellular matrix in the mesangium and tubulointerstitium. Recent studies suggest that the adult kidney also expresses type 2 (AT2 ) ANG II receptors in glomeruli, tubular segments, and vasculature. AT2 -receptor activation is associated with increased intrarenal nitric oxide production, stimulation of natriuresis, and inhibition of cell growth and matrix synthesis, effects that oppose those of kidney AT1 receptors. A number of studies have shown a reduction in kidney AT1 -receptor expression in diabetic nephropathy, suggesting that the balance between AT1 - and AT2 -receptor–mediated cell-signaling events may be a determinant of progression rate in diabetic nephropathy and that unopposed stimulation of AT2 receptors by ANG II with use of AT1 -receptor blockers may contribute to the beneficial properties of these agents. Determination of the expression pattern of AT2 receptors in diabetes and further definition of the role of AT2 receptors in opposing the detrimental effects of AT1 receptors may lead to more selective targeting of the RAS in diabetic nephropathy.

Down-Regulation of Renal Endothelial Nitric Oxide Synthase Expression in Experimental Glomerular Thrombotic Microangiopathy

Infection with certain strains of Escherichia coli and endotoxemia results in renal glomerular thrombotic microangiopathy (TMA) characterized by endothelial swelling and prominent glomerular microthrombus formation. Nitric oxide (NO) is an endogenous biologic modulator with diverse physiologic functions including vasodilation and inhibition of platelet adhesion and aggregation. NO is synthesized from conversion of L-arginine to L-citrulline by a family of NO synthases (NOS), which include constitutive and inducible isoforms. Indirect evidence supports the hypothesis that TMA is associated with depressed intrarenal NO production. However, the effect of TMA on renal tissue NOS expression has not been fully elucidated. We studied rats with TMA induced by iv bolus injection of high dose (20 mg/kg) E. coli endotoxin. Subgroups of six animals each were sacrificed before or at 30, 90, 180, 360, and 720 minutes after the administration of endotoxin. Renal histology and tissue expression of endothelial and inducible nitric oxide synthases (eNOS and iNOS) were examined. Additionally, we examined the effect of endotoxin on glomerular NO production, and eNOS and iNOS protein expression in vitro. Glomerular capillary thrombosis developed by 180 minutes after endotoxin administration in approximately half of the animals. The glomeruli without thrombotic lesions apparent by light microscopy disclosed early signs of TMA characterized by endothelial swelling, platelet accumulation/adhesion, and patchy fibrinogen deposition. These morphologic changes were associated with a marked reduction of renal tissue eNOS expression beyond 180 minutes after the endotoxin administration. The fall in eNOS expression was coupled with a significant rise in iNOS protein abundance, which was expressed largely by glomerular circulating neutrophils and endothelial cells, peritubular vascular endothelium, and collecting ducts of cortex and medulla. In vitro incubation of isolated glomeruli with endotoxin also resulted in a marked reduction in eNOS expression and a significant rise in iNOS content. Administration of E. coli endotoxin leads to a sustained fall in renal eNOS expression both in vivo and in vitro. The associated decline in intrarenal endothelial NO production/availability may result in renal vasoconstriction and a hypercoagulative state, which may contribute to the pathogenesis of endotoxin-induced TMA.

Responses to acute changes in arterial pressure on renal medullary nitric oxide activity in dogs.

A direct relationship between renal arterial pressure (RAP) and cortical tissue nitric oxide (NO) activity in the canine kidney was reported earlier. We have conducted further experiments to evaluate medullary NO responses to alterations in RAP with the use of a NO-selective microelectrode inserted into the renal medulla of 6 anesthetized, sodium-replete dogs. Graded reductions in RAP (from 140 to 80 mm Hg) elicited decreases in medullary tissue NO concentration, with a slope of 10.2+/-4.5 nmol x L(-1) x mm Hg(-1). These changes in NO levels were associated with decreases in urinary excretion rate of nitrate and nitrite (U(NOx)V; control value, 1.7+/-0.03 nmol x min(-1) x g(-1); slope, 0.02+/-0.004 nmol x min(-1) x g(-1) x mm Hg(-1)) and sodium excretion (U(Na)V; control, 3.2+/-0.7 micromol x min(-1) x g(-1); slope, 0.06+/-0.02 micromol x min(-1) x g(-1) x mm Hg(-1)) without changes in glomerular filtration rate control (0.84+/-0.06 mL x min(-1) x g(-1)). Intra-arterial administration of the NO synthase inhibitor N(omega)-nitro-L-arginine (NLA; 50 microg x kg(-1) x min(-1)) decreased medullary NO concentration by 218+/-55 nmol x L(-1) (n=5) and attenuated the relationship between RAP and NO concentration (slope, 2.7+/-2.2 nmol x L(-1) x mm Hg(-1)). NLA infusion decreased U(NOx)V (0.8+/-0.06 nmol x min(-1) x g(-1)) and U(Na)V (1.1+/-0.2 micromol x min(-1) x g(-1)) without changes in glomerular filtration rate and attenuated RAP versus U(Nox)V and U(Na)V relationships. Total and regional blood flows, as measured by electromagnetic and laser Doppler needle flow probes, respectively, remained autoregulated both before and during NLA infusion. These data support the hypothesis that acute changes in RAP elicit changes in intrarenal NO production, which may participate in the mediation of pressure natriuresis.

Nitric oxide metabolism following unilateral renal ischemia/reperfusion injury in rats.

Renal ischemia/reperfusion (I/R) injury results in decreased glomerular filtration and renal blood flow (RBF) and increased urine output, characterized by natriuresis and impaired concentrating ability. We studied unilateral I/R in rats to assess renal handling of nitric oxide (NO). Prior to I/ R, we measured urine flow rate (V), inulin clearance (C[IN]), para-aminohippuric acid clearance (C[PAH]), NO clearance (C[NOx] determined from metabolites NO2 and NO3), tubular transport of NOx (T[NOx], filtered load +/- urinary excretion), urine sodium and potassium excretion (U[Na]V, U[K]V), fractional excretion of sodium (FENa), and fractional excretion of NOx (FENOx) in each kidney. The left renal artery was then ligated for 30 min, followed by 30 min of reperfusion, and all measurements were repeated. C(IN) and C(PAH) were decreased in I/R kidneys compared with the contralateral kidney or pre-ischemia controls. V, FENa, and U(K)V were all significantly increased in I/R kidneys. Plasma NOx concentration was lower after injury in all animals (23.3 +/- 2.8 post injury vs. 30.4 +/- 7.7 microM pre injury, P < 0.05). C(NOx) was significantly higher in I/R kidneys (0.14 +/- 0.05 ml/min per g kidney weight) than in pre-injury kidneys (0.03 +/- 0.02 right, 0.04 +/- 0.30 left) or the contralateral controls (0.04 +/- 0.02) (P < 0.05 for all three controls). T(NOx) showed net tubular reabsorption of NOx in all kidneys (11 +/- 6 in post-ischemic left kidneys vs. 25 +/- 20 in left pre-ischemia, 33 +/- 13 in right pre-ischemia, and 21 +/- 4 right post-ischemia, nM/min per g kidney weight, P = NS). FENOx was higher in injured kidneys (28% +/- 18) than in pre-injury (3% +/- 0.6, 5% +/- 3) or contralateral controls (6% +/- 3) (P < 0.05 for all three controls). Renal NOx excretion and clearance are increased despite decreased plasma levels of NO metabolites after I/R injury. This increased excretion is not dependent on RBF or glomerular filtration, but may be related to impaired tubular reabsorption of NOx combined with increased intra-renal NO production.

NITRIC OXIDE IN THE MEDIATION OF PRESSURE NATRIURESIS

1. Recent studies have indicated that nitric oxide (NO) production in the kidney contributes to the regulation of renal haemodynamics and excretory function. Inhibition of nitric oxide synthase (NOS) reduces renal blood flow by approximately 25% and markedly reduces sodium excretion without reductions in filtered load. In particular, inhibition of NO synthesis markedly suppresses the slope of the arterial pressure-mediated response in sodium excretion. 2. Further studies have shown that constant intrarenal infusion of a NO donor in dogs treated with a NOS inhibitor produced diuretic and natriuretic responses but failed to restore the slope of the pressure-induced natriuretic response. These data indicate that an alteration in intrarenal NO activity, rather than the simple presence of NO during changes in arterial pressure is required for full expression of pressure natriuretic responses. 3. In support of the hypothesis that NO is involved in the mediation of pressure natriuresis, we also recently demonstrated a direct relationship between changes in arterial pressure and urinary excretion rate of sodium as well as nitrate and nitrite (a marker for endogenous NO activity) in the presence of efficient autoregulation of cortical and medullary blood flow. 4. The direct inhibitory actions of NO on tubular sodium reabsorption have also been observed in cultured tubular cells as well as isolated, perfused cortical collecting duct segments. 5. Thus, the collective data suggest that acute changes in arterial pressure induce changes in intrarenal NO production, which may directly alter tubular reabsorptive function to manifest the phenomenon of pressure natriuresis.