High NaCl Intake Research Articles

The Renin-Angiotensin and Renal Dopaminergic Systems Interact in Normotensive Humans.

The renin-angiotensin-aldosterone (RAAS) and renal dopaminergic systems interact to maintain sodium balance. High NaCl intake increases renal synthesis of dopamine and dopaminergic receptor activity, decreasing epithelial sodium transport, whereas sodium deficit activates the RAAS, increasing epithelial sodium transport. We tested the hypothesis that attenuation of the natriuretic effect of dopamine D1-like receptors during salt restriction results in part from increased RAAS activity in seven salt-resistant normotensive adults using a double-blind placebo-controlled balanced crossover design. All subjects attained sodium balance on low (50 mmol Na(+)/day) and high (300 mmol Na(+)/day) NaCl diets, administered 4 weeks apart. Sodium, potassium, lithium, para-aminohippurate, and creatinine clearances were measured before, during, and after a 3-hour infusion of fenoldopam, a D1-like receptor agonist, with and without pretreatment with enalapril, an angiotensin converting enzyme inhibitor. On the high NaCl diet, fenoldopam-induced natriuresis was associated with the inhibition of renal proximal and distal tubule sodium transport. On the low NaCl diet, fenoldopam decreased renal distal tubule sodium transport but did not cause natriuresis. The addition of enalapril to fenoldopam restored the natriuretic effect of fenoldopam and its inhibitory effect on proximal tubule sodium transport. Thus, on a high NaCl diet fenoldopam causes natriuresis by inhibiting renal proximal and distal tubule transport, but on a low NaCl diet the increased RAAS activity prevents the D1-like receptor from inhibiting renal proximal tubule sodium transport, neutralizing the natriuretic effect of fenoldopam. These results demonstrate an interaction between the renin-angiotensin and renal dopaminergic systems in humans and highlight the influence of dietary NaCl on these interactions.

Tumor necrosis factor‐alpha (TNF) regulates NKCC2 and AQP2 expression and adaptation to high NaCl intake

We previously showed that ambient urine osmolality was higher in TNF deficient mice compared with age‐ and sex‐matched wild type (WT) mice. As the role of TNF in the kidney is still being defined, effects of renal‐specific TNF silencing on renal Na(+)‐K(+)‐2Cl(‐) cotransporter (NKCC2) and aquaporin‐2 (AQP2) expression was determined. Six days after intrarenal injection of lentivirus construct EGFP‐TNF‐ex4 into both kidneys of WT mice, renal but not splenic TNF mRNA levels were significantly reduced. Injection of control lentivirus (U6) did not alter TNF mRNA in either kidney or spleen. EGFP‐TNF‐ex4, but not U6, also increased NKCC2A mRNA levels in renal outer medulla and inner medullary AQP2 mRNA and protein expression. Urine osmolality increased in mice injected with EGFP‐TNF‐ex4 compared with those injected with U6, suggesting that the increase in NKCC2 and AQP2 due to renal silencing of TNF contributed to an increase osmolality. As TNF production by the TAL was induced by administration of 1% NaCl in the drinking water for 3 days, the effects of TNF silencing on urine volume were determined. Mice were acclimated in metabolic cages for 3 days after receiving intrarenal injections of either EGFP‐TNF‐ex4 or U6. Food, body weight, water intake, and urine output was then measured in mice given 1% NaCl or tap water for 3 days. Both water intake and urine output were reduced in mice injected with EGFP‐TNF‐ex4 compared with mice given U6 lentivirus. The decrease in urine volume was associated with an increase in NKCC2A and AQP2 mRNA accumulation. Collectively, these findings suggest that TNF produced by the kidney inhibits transporter expression and regulates responses to high NaCl intake.

Abstract 14650: A Monoclonal Antibody to a Sodium Pump Inhibitor Marinobufagenin Reversed Aortic Remodeling and Stiffness in Normotensive Rats on a High Salt Intake

Background: Marinobufagenin (MBG), an endogenous cardiotonic steroid, is a Na/K-ATPase inhibitor and a vasoconstrictor. Previously it was demonstrated, that administration of 3E9 anti-MBG-antibody (mAb) reduced blood pressure (BP) and reversed left ventricular fibrosis in animal models of salt-sensitive hypertension and nephropathy. In the present study we investigated whether mAb alleviates BP and vascular remodeling in normotensive rats on a high salt intake. Methods: Wistar rats (5 months old) received normal salt diet (CTRL; n=8) or high salt intake (2% NaCl in drinking water) for 4 weeks. Rats on a high salt were administered vehicle (SALT; n=8) or mAb (50 ug/kg) (SALT-AB; n=8) 3 times during the last week of a high salt diet. BP was measured at baseline, after 3 and 4 weeks of experiment. Na/K-ATPase activity was measured in erythrocytes. Aortas were weighed, and were used to study sensitivity to the vasorelaxant effect of sodium nitroprusside (SNP), and for the histochemistry analysis of collagen deposition. Renal 24-hr MBG excretion was measured at week 4. Results: In SALT vs. CTRL, in the absence of BP changes, elevated levels of MBG (14.1±1.1 vs. 9.0±1.6 pmol/24hr, p<0.05) were associated with inhibition of erythrocyte Na/K-ATPase (12.6±0.3 vs. 14.2±0.35 μmol Pi/ml/hr, p<0.05), increased aortic weights (217±15 vs. 158±9 mg/kg BW, p<0.01), increased levels of collagen in aorta (2.5-fold; p<0.05), and compromised SNP vasorelaxant effect in aortic explants (EC50=167±19.3 nM vs. 99±2.0 nM; P<0.01). Antibody treatment in SALT-AB vs. SALT increased Na/K-ATPase activity (13.93±0.54 μmol Pi/ml/hr, p<0.05), reduced the aortic weight (180±12 mg/kg; P<0.05) and collagen deposition 3-fold (P<0.05), and restored the vasorelaxation of aortic rings by SNP to the levels in CTRL (70±1.5 nM, p<0.01). Conclusion: These findings for the first time demonstrated that in normotensive rats on a high salt intake heightened MBG levels induced vascular fibrosis and impairment of vasorelaxation in the absence of blood pressure changes. Immunoneutralization of MBG reversed these changes. Thus, high dietary NaCl intake in normotensive animals can stimulate vascular fibrosis via pressure-independent/ MBG-dependent mechanisms, and this remodeling is reversible.

Blood pressure, heart rate and tubuloglomerular feedback in A1AR-deficient mice with different genetic backgrounds.

Differences in genetic background between control mice and mice with targeted gene mutations have been recognized as a potential cause for phenotypic differences. In this study, we have used A1AR-deficient mice in a C57Bl/6 and SWR/J congenic background to assess the influence of background on the effect of A1AR-deficiency on cardiovascular and renal functional parameters. In A1AR+/+ and A1AR-/- mice in C57Bl/6 and SWR/J congenic backgrounds, we assessed blood pressure and heart rate using radio-telemetry, plasma renin concentrations and tubuloglomerular feedback. We did not detect significant differences in arterial blood pressure (MAP) and heart rates (HR) between A1AR+/+ and A1AR-/- mice in either C57Bl/6, SWR/J or mixed backgrounds. MAP and HR were significantly higher in SWR/J than in C57Bl/6 mice. A high NaCl intake increased MAP in A1AR-/- mice on C57Bl/6 background while there was less or no salt sensitivity in the SWR/J background. No significant differences in plasma renin concentration were detected between A1AR-/- and A1AR+/+ mice in any of the strains. Tubuloglomerular feedback was found to be absent in A1AR-/- mice with SWR/J genetic background. While this study confirmed important differences between inbred mouse strains, we did not identify phenotypic modifications of A1AR-related effects on blood pressure, heart rate and plasma renin by differences in genetic background.

Short term effects of increasing dietary salt concentrations on urine composition in healthy cats

High dietary salt (NaCl) concentrations are assumed to be beneficial in preventing the formation of calcium oxalate (CaOx) uroliths in cats, since increased water intake and urine volume have been observed subsequent to intake. In human beings, dietary NaCl restriction is recommended for the prevention of CaOx urolith formation, since high NaCl intake is associated with increased urinary Ca excretion. The aim of the present study was to clarify the role of dietary NaCl in the formation of CaOx uroliths in cats. Eight cats received four diets that differed in Na and Cl concentrations (0.38–1.43% Na and 0.56–2.52% Cl dry matter, DM). Each feeding period consisted of a 21 day adaptation period, followed by a 7 day sampling period for urine collection. Higher dietary NaCl concentrations were associated with increased urine volume and renal Na excretion. Urinary Ca concentration was constant, but renal Ca excretion increased from 0.62 to 1.05 mg/kg bodyweight (BW)/day with higher dietary NaCl concentrations (P ≤ 0.05). Urinary oxalate (Ox), citrate, P and K concentrations decreased when NaCl intake was high (P ≤ 0.05), and urinary pH was low in all groups (6.33–6.45; P > 0.05). Relative supersaturation of CaOx in the urine was unaffected by dietary NaCl concentrations. In conclusion, the present study demonstrated several beneficial effects of high dietary NaCl intake over a relatively short time period. In particular, urinary Ca concentration remained unchanged because of increased urine volume. Decreased urinary Ox concentrations might help to prevent the formation of CaOx uroliths, but this should be verified in future studies in diseased or predisposed cats.

High salt intake aggravates aortic dysfunction in rats with adenine‐induced chronic renal failure (1136.13)

Rats with adenine‐induced chronic renal failure (ACRF) develop a reduced rate of relaxation, and increased stiffness of the aorta. The aim of this study was to investigate the effects of 2 weeks of high NaCl intake on vascular function in rats with ACRF.Male Sprague‐Dawley rats received either chow containing adenine (ACRF) or were pair‐fed a control diet (Controls, C). Two weeks prior to sacrifice animals were randomized to diet containing either normal salt (0.6% NaCl, NS) or high salt (4% NaCl, HS), resulting in four groups: C‐NS, C‐HS, ACRF‐NS and ACRF‐HS (n=11‐15 per group). Animals were sacrificed at 7‐10 weeks after study start and mesenteric arteries and thoracic aorta were analyzed with wire myograph.Animals with ACRF showed significant increases in serum creatinine and systolic blood pressure that was aggravated by high NaCl intake (138±14, 164±10, 152±7 and 202±14 mmHg in groups C‐NS, ACRF‐NS, C‐HS and ACRF‐HS, respectively). A decreased sensitivity to norepinephrine was found in mesenteric arteries of ACRF‐HS rats (P<0.05). In the thoracic aorta, the sensitivity to both sodium nitroprusside (SNP) and acetylcholine (ACh) were reduced only in ACRF‐HS animals (P<0.05). Furthermore, the ACRF‐HS group exhibited a marked decrease in aortic relaxation rate compared to ACRF‐NS animals (P<0.01).In rats with ACRF two weeks of high NaCl intake reduced the sensitivity to ACh and SNP in the thoracic aorta and aggravated the impairment in relaxation rate whereas vasodilator responses were unaffected in resistance arteries. These results indicate an abnormality in vascular smooth muscle function specifically in the thoracic aorta of ACRF animals that is augmented by high NaCl intake.

Prostaglandin E2 induces chloride secretion through crosstalk between cAMP and calcium signaling in mouse inner medullary collecting duct cells

Under conditions of high dietary salt intake, prostaglandin E2 (PGE2) production is increased in the collecting duct and promotes urinary sodium chloride (NaCl) excretion; however, the molecular mechanisms by which PGE2 increases NaCl excretion in this context have not been clearly defined. We used the mouse inner medullary collecting duct (mIMCD)-K2 cell line to characterize mechanisms underlying PGE2-regulated NaCl transport. When epithelial Na(+) channels were inhibited, PGE2 exclusively stimulated basolateral EP4 receptors to increase short-circuit current (Isc(PGE2)). We found that Isc(PGE2) was sensitive to inhibition by H-89 and CFTR-172, indicating that EP4 receptors signal through protein kinase A to induce Cl(-) secretion via cystic fibrosis transmembrane conductance regulator (CFTR). Unexpectedly, we also found that Isc(PGE2) was sensitive to inhibition by BAPTA-AM (Ca(2+) chelator), 2-aminoethoxydiphenyl borate (2-APB) (inositol triphosphate receptor blocker), and flufenamic acid (FFA) [Ca(2+)-activated Cl(-) channel (CACC) inhibitor], suggesting that EP4 receptors also signal through Ca(2+) to induce Cl(-) secretion via CACC. Additionally, we observed that PGE2 stimulated an increase in Isc through crosstalk between cAMP and Ca(2+) signaling; BAPTA-AM or 2-APB inhibited a component of Isc(PGE2) that was sensitive to CFTR-172 inhibition; H-89 inhibited a component of Isc(PGE2) that was sensitive to FFA inhibition. Together, our findings indicate that PGE2 activates basolateral EP4 receptors and signals through both cAMP and Ca(2+) to stimulate Cl(-) secretion in IMCD-K2 cells. We propose that these signaling pathways, and the crosstalk between them, may provide a concerted mechanism for enhancing urinary NaCl excretion under conditions of high dietary NaCl intake.

Deletion of cyclooxygenase-2 in the mouse increases arterial blood pressure with no impairment in renal NO production in response to chronic high salt intake

Experiments were designed to test the hypothesis that cyclooxygenase-2 (COX-2) activity attenuates the blood pressure increase during high NaCl intake by stimulation of endothelial nitric oxide synthase (eNOS)-mediated NO synthesis in the kidney medulla. COX-2(-/-) (C57BL6) an COX-2(+/+) mice were fed a diet with 0.004% (low salt, LS) or 4% (high salt, HS) NaCl for 18 days. Arterial blood pressure was recorded continuously using indwelling catheters. Food and water intake and diuresis were measured in metabolic cages. Urine osmolality and excretion of electrolytes, cGMP, cAMP, and NOx were determined, as well as plasma NOx and cGMP. There was a significant dependence of blood pressure on salt intake and genotype: COX-2(-/-) exhibited higher blood pressure than COX-2(+/+) both on HS and LS intake. COX-2(+/+) littermates displayed an increase in blood pressure on HS versus LS (102.3 ± 1.1 mmHg vs. 91.9 ± 0.9 mmHg) day and night. The mice exhibited significant blood pressure increases during the awake phase (night) that were larger in COX-2(-/-) on HS diet compared with COX-2(+/+). Water intake, diuresis, Na(+), and osmolyte excretions and NOx and cGMP excretions were significantly and similarly elevated with HS in COX-2(-/-) and COX-2(+/+). In summary, C57BL6 mice exhibit a salt intake-dependent increase in arterial blood pressure with increased renal NO production. COX-2 activity has a general lowering effect on arterial blood pressure. COX-2 dampens NaCl-induced increases in arterial blood pressure in the awake phase. In conclusion, COX-2 activity attenuates the changes in nocturnal blood pressure during high salt intake, and COX-2 activity is not necessary for increased renal nitric oxide formation during elevated NaCl intake.

Effects of high dietary sodium chloride content on performance and sodium and potassium balance in growing pigs

Thirty castrated male Moo Lath pigs (6-8 weeks of age) were used in a 15-week growth trial to study the effect of high dietary sodium chloride (NaCl) content on feed and water intake, performance, sodium (Na) and potassium (K) balance, and plasma aldosterone concentration. The pigs were randomly allocated (ten per treatment) to diets containing 0.24 % Na (Na0.24), 0.28 % Na (Na0.28), and 0.32 % Na (Na0.32) per kg diet. Feed and water was provided ad libitum, and water consumption, feed offered, and feed residues were recorded daily. Every third week, the pigs were weighed, blood samples were collected, and a 3-day total collection of urine and feces was performed. Water intake was higher (P = 0.001) in pigs fed with diets Na0.28 (3.7 L/day) and Na0.32 (3.9 L/day) than in pigs fed with diet Na0.24 (3.4 L/day), and dry matter (DM) intake was higher on diet Na0.32 (P = 0.041) than on the other diets. The average daily body weight (BW) gain was higher on diet Na0.32 than on the other diets (P = 0.031). The feed conversion ratio (in kilogram feed DM to kilogram BW gain) was 4.6, 4.6, and 4.1 on treatments Na0.24, Na0.28, and Na0.32, respectively (P = 0.14). The highest Na balance was observed on diet Na0.32 followed by diets Na0.28 and Na0.24 (P < 0.001), while there was no treatment-related pattern for the K balance. The Na/K ratio in feces and urine increased (P < 0.001), and the K/Na ratio in feces (P < 0.001) decreased with increasing Na content in the diet. Plasma aldosterone concentration decreased (P < 0.001) with increasing dietary content of Na. These results indicate that high NaCl intake and free access to water will increase Na balance but do not negatively influence feed intake and performance of growing local pigs.

Alkaline Salts to Counteract Bone Resorption and Protein Wasting Induced by High Salt Intake: Results of a Randomized Controlled Trial

High sodium chloride (NaCl) intake can induce low-grade metabolic acidosis (LGMA) and may thus influence bone and protein metabolism. We hypothesized that oral potassium bicarbonate (KHCO(3)) supplementation may compensate for NaCl-induced, LGMA-associated bone resorption and protein losses. Eight healthy male subjects participated in a randomized trial with a crossover design. Each of two study campaigns consisted of 5 d of dietary and environmental adaptation followed by 10 d of intervention and 1.5 d of recovery. In one study campaign, 90 mmol KHCO(3)/d were supplemented to counteract NaCl-induced LGMA, whereas the other campaign served as a control with only high NaCl intake. When KHCO(3) was ingested during high NaCl intake, postprandial buffer capacity ([HCO(3)(-)]) increased (P = 0.002). Concomitantly, urinary excretion of free potentially bioactive glucocorticoids [urinary free cortisol (UFF) and urinary free cortisone (UFE)] was reduced by 14% [∑(UFF,UFE); P = 0.024]. Urinary excretion of calcium and bone resorption marker N-terminal telopeptide of type I collagen was reduced by 12 and 8%, respectively (calcium, P = 0.047; N-terminal bone collagen telopeptide, P = 0.044). There was a trend of declining net protein catabolism when high NaCl was combined with KHCO(3) (P = 0.052). We conclude that during high salt intake, the KHCO(3)-induced postprandial shift to a more alkaline state reduces metabolic stress. This leads to decreased bone resorption and protein degradation, which in turn might initiate an anticatabolic state for the musculoskeletal system in the long run.

NaCl Potentiates Human Fibrocyte Differentiation

Excessive NaCl intake is associated with a variety of fibrosing diseases such as renal and cardiac fibrosis. This association has been attributed to increased blood pressure as the result of high NaCl intake. However, studies in patients with high NaCl intake and fibrosis reveal a connection between NaCl intake and fibrosis that is independent of blood pressure. We find that increasing the extracellular concentration of NaCl to levels that may occur in human blood after high-salt intake can potentiate, in serum-free culture conditions, the differentiation of freshly-isolated human monocytes into fibroblast-like cells called fibrocytes. NaCl affects the monocytes directly during their adhesion. Potassium chloride and sodium nitrate also potentiate fibrocyte differentiation. The plasma protein Serum Amyloid P (SAP) inhibits fibrocyte differentiation. High levels of extracellular NaCl change the SAP Hill coefficient from 1.7 to 0.8, and cause a four-fold increase in the concentration of SAP needed to inhibit fibrocyte differentiation by 95%. Together, our data suggest that NaCl potentiates fibrocyte differentiation. NaCl-increased fibrocyte differentiation may thus contribute to NaCl-increased renal and cardiac fibrosis.

Effects of Endothelin Receptor Antagonists on Renal Hemodynamics in Angiotensin II-Infused Rats on High NaCl Intake

Aim: The aim was to investigate effects of selective endothelin (ET) receptor antagonists on renal hemodynamics and dynamic renal blood flow autoregulation (RBFA) in angiotensin II (Ang II)-infused rats on a high NaCl intake. Methods: Sprague-Dawley rats received Ang II (250 ng/kg/min, s.c.) and an 8 % NaCl diet for 14 days after which renal clearance experiments were performed. After baseline measurements animals were administered either: (a) saline vehicle; (b) ET_A receptor antagonist BQ-123 (30 nmol/kg/min); (c) ET_B receptor antagonist BQ-788 (30 nmol/kg/min); or (d) BQ-123 + BQ-788, for six consecutive 20-minute clearance periods. Results: BQ-123 reduced arterial pressure (AP) and selectively increased outer medullary perfusion versus vehicle (p<0.05). These effects were attenuated or abolished by combined BQ-123 and BQ-788. BQ-788 reduced renal blood flow and increased renovascular resistance (p<0.05). Ang II-infused rats on high NaCl intake showed abnormalities in dynamic RBFA characterized by an impaired myogenic response that were not significantly affected by ET receptor antagonists. Conclusion: In hypertensive Ang II-infused rats on a high-NaCl intake selective ET_A antagonism with BQ-123 reduced AP and specifically increased OM perfusion and these effects were dependent on intact ET_B receptor stimulation. Furthermore, ET receptor antagonists did not attenuate abnormalities in dynamic RBFA.

High sodium chloride intake exacerbates immobilization-induced bone resorption and protein losses

We examined, in immobilization, the effect of a diet high in sodium chloride (NaCl) on bone markers, nitrogen balance, and acid-base status. Eight healthy male test subjects participated in a 14-day head-down-tilt bed rest (HDBR) study. During the bed rest period they received, in a randomized crossover design, a high (7.7 meq Na(+)/kg body wt per day) and a low (0.7 meq Na(+)/kg body wt per day) NaCl diet. As expected, 24-h excretion of urinary calcium was significantly greater in the high-NaCl-intake HDBR phase than in the low-NaCl-intake HDBR phase (P < 0.001). High NaCl intake caused a 43-50% greater excretion of the bone resorption markers COOH- (CTX) and NH(2)- (NTX) terminal telopeptide of type I collagen in HDBR than low NaCl in HDBR (CTX/NTX: P < 0.001). Serum concentrations of the bone formation markers bone-specific alkaline phosphatase (bAP) and NH(2)-terminal propeptide of type I procollagen (PINP) were identical in both NaCl intake phases. High NaCl intake led to a more negative nitrogen balance in HDBR (P < 0.001). Changes were accompanied by increased serum chloride concentration (P = 0.008), reduced blood bicarbonate (P = 0.017), and base excess (P = 0.009) whereas net acid excretion was lower during high than during low NaCl intake in immobilization (P < 0.001). High NaCl intake during immobilization exacerbates disuse-induced bone and muscle loss by causing further protein wasting and an increase in bone resorption. Changes in the acid-base status, mainly caused by disturbances in electrolyte metabolism, seem to determine NaCl-induced degradation processes.

Infiltrating T lymphocytes in the kidney increase oxidative stress and participate in the development of hypertension and renal disease

The present studies examined the role and mechanism of action of infiltrating T lymphocytes in the kidney during salt-sensitive hypertension. Infiltrating T lymphocytes in the Dahl salt-sensitive (SS) kidney significantly increased from 7.2 ± 1.8 × 10(5) cells/2 kidneys to 18.2 ± 3.9 × 10(5) cells/2 kidneys (n = 6/group) when dietary NaCl was increased from 0.4 to 4.0%. Furthermore, the expression of immunoreactive p67(phox), gp91(phox), and p47(phox) subunits of NADPH oxidase was increased in T cells isolated from the kidneys of rats fed 4.0% NaCl. The urinary excretion of thiobarbituric acid-reactive substances (TBARS; an index of oxidative stress) also increased from 367 ± 49 to 688 ± 92 nmol/day (n = 8/group) when NaCl intake was increased in Dahl SS rats. Studies were then performed on rats treated with a daily injection of vehicle (5% dextrose) or tacrolimus (0.25 mg·kg(-1)·day(-1) ip), a calcineurin inhibitor that suppresses immune function, during the period of high-NaCl intake (n = 5/group). In contrast to the immune cell infiltration, increased NADPH oxidase expression, and elevated urine TBARS excretion in vehicle-treated Dahl SS fed high salt, these parameters were unaltered as NaCl intake was increased in Dahl SS rats administered tacrolimus. Moreover, tacrolimus treatment blunted high-salt mean arterial blood pressure and albumin excretion rate (152 ± 3 mmHg and 20 ± 9 mg/day, respectively) compared with values in dextrose-treated Dahl SS rats (171 ± 8 mmHg and 74 ± 28 mg/day). These experiments indicate that blockade of infiltrating immune cells is associated with decreased oxidative stress, an attenuation of hypertension, and a reduction of renal damage in Dahl SS rats fed high salt.

Vitamin B 6 and Oxalic Acid in Clinical Nephrology

Vitamin B(6) (VB(6)) is a water-soluble vitamin, which is important for the normal functioning of multiple organ systems. It is metabolized to the active molecule pyridoxal-5-phosphate (PLP). Oxalic acid (OA) is thought to be a uremic toxin that participates in the pathogenesis of the uremic syndrome. The objectives of this study were as follows: (1) to evaluate the plasma and erythrocyte VB(6) (effect of PLP; effect of PLP was in indirect relationship with the concentration of erythrocyte VB(6)), and plasma and urinary OA in marathon runners, in patients with acute intermittent porphyria (AIP) and variegate porphyria, and in patients with stage 1 chronic kidney disease (CKD), chronic glomerulonephritis and nephrotic syndrome (CGNS); (2) to examine the influence of water diuresis in healthy subjects, and the influence of sodium diuresis (high sodium intake) and an intravenous administration of furosemide on the urinary excretion of VB(6) and OA in CKD stage 3-4 patients; and (3) to evaluate the influence of erythropoietin treatment on erythrocyte VB(6) (effect of PLP) in hemodialysis (HD) patients, and the influence of continuous ambulatory peritoneal dialysis (CAPD) therapy on plasma VB(6) and OA and their peritoneal clearance and transfer. This study was conducted at the Nephrological Clinic of L. Pasteur Faculty Hospital and of Medical School of P. J. Safarik University. A combination of 29 marathon runners, 15 patients with CG and NS, 11 patients with AIP, 1 patient with variegate porphyria, 15 healthy subjects, 27 CKD stage 3-4 patients, 30 HD, and 27 CAPD patients were used in the study. After a marathon run, plasma and erythrocyte VB(6) significantly decreased and plasma OA increased. Plasma (15.5 +/- 3.8 nmol/L) and erythrocyte VB(6) (effect of PLP: 42.1% +/- 7.5%) were decreased and plasma OA (9.8 +/- 2.3 micromol/L) was significantly elevated in patients with CGNS and stage 1 CKD. In patients with AIP, deficiency of plasma (24.3 +/- 5.2 nmol/L) and erythrocyte VB(6) (effect of PLP: 46.2% +/- 7.0%) and hyperoxalemia (9.39 +/- 2.5 micromol/L) were present. The urinary excretion of VB(6) and of OA during maximal water diuresis and after intravenous administration of furosemide increased significantly (P < .01), but was not affected by the high intake of NaCl (P > .05). Erythropoietin treatment in HD patients led to the erythrocyte VB(6) deficiency. This finding is an indirect evidence that erythrocyte VB(6) is consumed by the hemoglobin synthesis much more during EPO treatment. In CAPD patients, plasma value of VB(6) (127.3 +/- 66.9 micromol/L) was in the normal range and plasma OA (23.6 +/- 7.4 micromol/L) was significantly elevated. Mean value of peritoneal clearance of VB(6) was 8.8% and of OA was 76.9% of urea clearance. Our study indicates that deficiency of VB(6) led to hyperoxalemia and hyperoxaluria in patients with CKD. Deficiency of VB(6) in CKD stage 4-5 patients potentiates the uremic hyperoxalemia and hyperoxaluria.

Sodium, Potassium, and Target Organ Damage

The relationship between blood pressure (BP) and sodium intake is well documented. Large observational studies have shown that sodium chloride (NaCl) intake is directly related to BP, whereas various interventional studies demonstrated a relationship between the reduction in NaCl intake and magnitude of BP reduction. Similarly, a low-potassium diet is associated with high BP and increased cardiovascular risk. In the Intersalt Study, urinary potassium excretion (a surrogate for intake) was associated with a lower BP after adjustment for various confounders, including urinary sodium excretion. The effects of dietary sodium and potassium on BP are closely related. The first strain of rats bred by Dahl for the trait of “salt sensitivity” demonstrated less NaCl-induced hypertension when potassium intake was high. Well-controlled human studies subsequently showed that the BP-rising effect of extremely high NaCl intake can be blunted by potassium supplementation. Clinical trials showed that potassium supplementation modestly reduces BP, an effect that is, however, enhanced in blacks and in the presence of a high-sodium diet. Conversely, hypertension in potassium deficiency is associated with increased renal sodium reabsorption, whereas sodium restriction may prevent the development of hypertension induced by potassium restriction. Recent literature has identified regulatory pathways of tubular sodium and potassium handling (eg, those mediated by with-no-lysine-kinase 1)1 that may, at least partially, mediate interactions between dietary sodium and potassium on extracellular volume and BP. Importantly, whereas high-sodium/low-potassium intake are associated with hypertension, both have been shown to predict an increased cardiovascular risk even after adjustment for brachial BP and other observed factors, indicating that unmeasured factors played a role. Accumulating evidence indicates that central arterial pulsatile hemodynamic abnormalities predict cardiovascular risk independent of brachial BP in various populations.2 It should be noted that there is a highly variable relationship between brachial pulse pressure (PP) and aortic pulsatile hemodynamics. …

High-NaCl intake impairs dynamic autoregulation of renal blood flow in ANG II-infused rats

The aim of this study was to investigate dynamic autoregulation of renal blood flow (RBF) in ANG II-infused rats and the influence of high-NaCl intake. Sprague-Dawley rats received ANG II (250 ng·kg(-1)·min(-1) sc) or saline vehicle (sham) for 14 days after which acute renal clearance experiments were performed during thiobutabarbital anesthesia. Rats (n = 8-10 per group) were either on a normal (NNa; 0.4% NaCl)- or high (HNa; 8% NaCl)-NaCl diet. Separate groups were treated with 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (tempol; 1 M in drinking water). Transfer function analysis from arterial pressure to RBF in the frequency domain was used to examine the myogenic response (MR; 0.06-0.09 Hz) and the tubuloglomerular feedback mechanism (TGF; 0.03-0.06 Hz). MAP was elevated in ANG II-infused rats compared with sham groups (P < 0.05). RBF in ANG II HNa was reduced vs. sham NNa and sham HNa (6.0 ± 0.3 vs. 7.9 ± 0.3 and 9.1 ± 0.3 ml·min(-1)·g kidney wt(-1), P < 0.05). transfer function gain in ANG II HNa was significantly elevated in the frequency range of the MR (1.26 ± 0.50 dB, P < 0.05 vs. all other groups) and in the frequency range of the TGF (-0.02 ± 0.50 dB, P < 0.05 vs. sham NNa and sham HNa). Gain values in the frequency range of the MR and TGF were significantly reduced by tempol in ANG II-infused rats on HNa diet. In summary, the MR and TGF components of RBF autoregulation were impaired in ANG II HNa, and these abnormalities were attenuated by tempol, suggesting a pathogenetic role for superoxide in the impaired RBF autoregulatory response.

In vivo stimulation of AMP-activated protein kinase enhanced tubuloglomerular feedback but reduced tubular sodium transport during high dietary NaCl intake

AMP-activated protein kinase (AMPK) is expressed in the apical membrane of cortical thick ascending limb, distal, and collecting tubules as well as macula densa cells of the kidneys. AMPK is an active modulator of epithelial Na(+) channels, Na(+)-2Cl(-)-K(+) cotransporter, and the ATP-dependent potassium channel. The present experiments explored whether AMPK participates in the regulation of tubuloglomerular feedback (TGF) and renal tubular sodium handling. To this end, renal clearance and micropuncture experiments were performed in anesthetized rats. Under normal NaCl diet, neither TGF response nor renal fluid and sodium excretion were altered by pharmacological activation of AMPK in vivo. However, under high NaCl diet, the TGF response was significantly enhanced after intravenous or intratubular application of the AMPK activator AICAR. Moreover, AICAR application significantly increased fractional delivery of fluid and sodium to the end of the proximal tubule. High dietary NaCl intake increased the renal transcript levels encoding the AMPK-alpha1 subunit, while it decreased the expression of AMPK-beta1 and AMPK-gamma2 subunits. Immunoblots revealed that high dietary NaCl intake reduced renal expression of activated AMPK by about three times compared to normal NaCl diet whereas additional AICAR application increased AMPK activity. Our results suggest that AMPK regulates tubuloglomerular balance as well as tubular transport upon change of renal work load.

Adenosine Activates A2b Receptors and Enhances Chloride Secretion in Kidney Inner Medullary Collecting Duct Cells

In the kidney, defects in the regulation of urine salt excretion can result in extracellular fluid volume expansion, leading to salt-sensitive hypertension. Previous studies have demonstrated that, when rats are maintained on a high sodium chloride (NaCl) diet, adenosine production increases in the renal medulla with parallel changes in adenosine receptor expression. These studies suggest that adenosine signaling in the kidney can respond to high NaCl loading; however, the functional consequences of these changes in adenosine signaling are not clear. We used the immortalized cell line mIMCD-K2, a murine model system for the renal inner medullary collecting duct, to study the direct effects of adenosine on NaCl transport across the inner medullary collecting duct epithelium with an Ussing chamber system. When epithelial Na(+) channels were inhibited, the addition of adenosine to the apical side of mIMCD-K2 cell sheets stimulated short-circuit current in a dose-dependent manner. This increase in short-circuit current was inhibited by a cystic fibrosis transmembrane conductance regulator Cl(-) channel inhibitor. Pharmacological studies with a panel of adenosine receptor agonists and antagonists demonstrated that adenosine activates apical A2b adenosine receptors to enhance the short-circuit current. Furthermore, adenosine application to mIMCD-K2 cell sheets increased intracellular cAMP, whereas inhibition of protein kinase A completely blocked the adenosine response. Together, our findings indicate that adenosine stimulates Cl(-) secretion through the cystic fibrosis transmembrane conductance regulator in mIMCD-K2 cells by activating apical A2b receptors and signaling through cAMP/protein kinase A. We propose that this adenosine receptor pathway may provide one mechanism for enhancing urine NaCl excretion in the setting of high dietary NaCl intake.

Mineral metabolism of sheep fed saltbush or a formulated high-salt diet

Old man saltbush ( Atriplex nummularia) is sown in saline farming systems worldwide, and is commonly utilised as fodder for livestock. Saltbush contains very high concentrations of NaCl and KCl, however, the complex interactions between minerals means that sheep grazing saltbush may be susceptible to mineral deficiencies. The aim of this experiment was to assess the effect of high NaCl and KCl intakes on the apparent absorption of some macro-minerals (Mg, Ca, P) by sheep fed formulated high-salt diets or old man saltbush to help identify likely mineral deficiencies. Two experiments were conducted. In both experiments 18-month-old Merino wethers were offered one of 4 diets at maintenance: a control pellet (no added salt), control pellet + 10% salt (low salt), control pellet + 20% salt (high salt), or old man saltbush containing approximately 20% salt. The salt added to all diets was 2NaCl:1KCl. Sheep were fed the diets for a minimum of 3 weeks before any measurements were taken. In the first experiment we measured the accumulation of Na and K in the rumen of cannulated sheep for 24 h following feeding. In the second experiment we measured the apparent digestion and absorption of Na, K, Cl, Mg, Ca and P over 6 days. There was a net loss of Mg (0.83 g/day), Ca (0.61 g/day) and P (0.46 g/day) from sheep fed the saltbush diet, but the addition of salt to the control pellet and high levels of Na and K in the rumen had no effect on the apparent digestion and absorption of Mg, Ca and P. The loss of Mg, Ca and P from the gastrointestinal tract of sheep fed saltbush is likely to be due to a combination of reduced availability and absorption of minerals from the saltbush leaves. Deficiencies in Mg, Ca and P can reduce animal production and even cause stock deaths. These results indicate that saltbush as a sole source of feed may be unsuitable for sheep with high nutritional demands, and further research is required to fully assess the mineral balance of sheep grazing saltbush pastures.