Diferencias de género en presión arterial, función renal y respuesta a la dieta hipersódica en ratas Wistar / Gender differences in blood pressure, renal function and response to a high-sodium diet in Wistar rats

Introducción: Es conocido que el sexo es un condicionante de la regulación renal de sodio y de la presión arterial. Material y métodos: Se estudiaron ratas Wistar machos y hembras a los 150 días de vida, con dieta normo o hipersódica (NaCl 1% v.o.) en los últimos cinco. Se determinaron presión arterial media (PAM), natriuresis, filtrado glomerular (VFG), flujo plasmático renal (FPR) y aldosterona plasmática. Se estudió la expresión Na+,K+-ATPasa total (t-NKA) y defosforilada (d-NKA), citocromo P4504A (CYP4A), cotransportadores Na+,K+,2Cl- tipo 2 (NKCC2) y Na+/Cl- (NCC) y por PCR el ARNm de la cadena α1 de NKA (Atp1a1) en corteza y médula renal. Resultados: La PAM fue mayor y la natriuresis menor en los machos bajo ambas dietas. Con ingesta hipersódica la aldosterona bajó en ambos sexos, el VFG fue menor en hembras y el FPR aumentó en machos (4,09 ± 0,17 vs 2,81 ± 0,12 ml/min/gR; p<0,01 vs dieta normosódica). La t-NKA, d-NKA y Atp1a1 en médula fue mayor en machos con ambas dietas. Con ingesta hipersódica, t-NKA en médula y d-NKA en corteza y médula disminuyeron en hembras y solamente d-NKA disminuyó en médula de machos. Asimismo, aumentó CYP4A y disminuyó NKCC2 y NCC en hembras, mientras que aumentó NKCC2, sin cambios en NCC, en machos. Conclusión: El sexo condiciona la presión arterial y el balance de sodio, disminuyendo su reabsorción en hembras y aumentando el FPR en machos. Esto sugiere posibilidades de estudio diferenciales según sexo en trastornos del metabolismo del sodio

Introduction: It is known that sex is a determinant of renal sodium regulation and blood pressure. Methods: Male and female Wistar rats, which were 150 days old and a diet with normal or high levels of sodium (NaCl 1% v.o.), were studied for the last five days. Mean blood pressure (MBP), natriuresis, glomerular filtration rate (GFR), renal plasma flow (RPF) and plasma aldosterone level were established. The following were studied: expressions of total Na+,K+,-ATPase (t-NKA); dephosphorylated NKA (d-NKA); cytochrome P4504A (CYP4A); Na+K+-2Cl- (NKCC2) and Na+/Cl- (NCC) cotransporters. The mRNA expression of the NKA α1 (Atp1a1) chain was examined through PCR analysis in the renal cortex and marrow. Results: Male rats having both types of diet showed higher MBP and lower natriuresis. High sodium intake triggered lower aldosterone levels in both sexes; GFR was lower in females and RPF was higher in males (4.09 ± 0.17 vs. 2.81 ± 0.12 ml/min/gr; p<0.01 vs. diet with a normal sodium level). Marrow t-NKA, d-NKA and Atp1a1 were higher in males on both diets. High sodium intake caused lower marrow t-NKA as well as lower cortex and marrow d-NKA in females. In the case of males, only marrow d-NKA decreased. Furthermore, females showed a higher level of CYP4A and lower levels of NKCC2 and NCC, whereas males showed higher levels of NKCC2 and no variations in NCC. Conclusion: Sex conditions blood pressure and sodium balance, reducing resorption in females and increasing RPF in males. This suggests the possibility of studying sodium metabolism disorders differently according to sex

High Dose Vitamin D3 Attenuates the Hypocalciuric Effect of Thiazide in Hypercalciuric Rats

Thiazide is known to decrease urinary calcium excretion. We hypothesized that thiazide shows different hypocalciuric effects depending on the stimuli causing hypercalciuria. The hypocalciuric effect of hydrochlorothiazide (HCTZ) and the expression of transient receptor potential vanilloid 5 (TRPV5), calbindin-D(28K), and several sodium transporters were assessed in hypercalciuric rats induced by high calcium diet and vitamin D3. Urine calcium excretion and the expression of transporters were measured from 4 groups of Sprague-Dawley rats; control, HCTZ, high calcium-vitamin D, and high calcium-vitamin D with HCTZ groups. HCTZ decreased urinary calcium excretion by 51.4% in the HCTZ group and only 15% in the high calcium-vitamin D with HCTZ group. TRPV5 protein abundance was not changed by HCTZ in the high calcium-vitamin D with HCTZ group compared to the high calcium-vitamin D group. Protein abundance of NHE3, SGLT1, and NKCC2 decreased in the hypercalciuric rats, and only SGLT1 protein abundance was increased by HCTZ in the hypercalciuric rats. The hypocalciuric effect of HCTZ is attenuated in high calcium and vitamin D-induced hypercalciuric rats. This attenuation seems to have resulted from the lack of HCTZ's effect on protein abundance of TRPV5 in severe hypercalciuric condition induced by high calcium and vitamin D.

Decreased Expression of Na+/K+-ATPase, NHE3, NBC1, AQP1 and OAT in Gentamicin-induced Nephropathy

The present study was aimed to determine whether there is an altered regulation of tubular transporters in gentamicin-induced nephropathy. Sprague-Dawley male rats (200~250 g) were subcutaneously injected with gentamicin (100 mg/kg per day) for 7 days, and the expression of tubular transporters was determined by immunoblotting and immunohistochemistry. The mRNA and protein expression of OAT was also determined. Gentamicin-treated rats exhibited significantly decreased creatinine clearance along with increased plasma creatinine levels. Accordingly, the fractional excretion of sodium increased. Urine volume was increased, while urine osmolality and free water reabsorption were decreased. Immunoblotting and immunohistochemistry revealed decreased expression of Na+/K+-ATPase, NHE3, NBC1, and AQP1 in the kidney of gentamicin-treated rats. The expression of OAT1 and OAT3 was also decreased. Gentamicin-induced nephropathy may at least in part be causally related with a decreased expression of Na+/K+-ATPase, NHE3, NBC1, AQP1 and OAT.

Altered Expression of Sodium Transporters and Water Channels in the Submandibular Gland of Rats Treated with Nitric Oxide Synthesis Inhibitors

A role of nitric oxide (NO) in the regulation of sodium transporters and water channels in the salivary gland was investigated. Male Sprague-Dawley rats were treated with NG-nitro-L-arginine methyl ester (L- NAME, 100 mg/L drinking water) for 1 week. The control group was supplied with normal tap water. The expression of Na+,K+-ATPase, type 2 Na+/K+/2Cl- cotransporter (NKCC2), type 1 Na+/H+ exchanger (NHE1), alpha-subunit of epithelial sodium transporter (ENaC), and aquaporin-5 (AQP5) and aquaporin-1 (AQP1) proteins were determined in the submandibular gland by Western blot analysis. Following the treatment with L-NAME, the expression of Na+,K+-ATPase alpha1-subunit, NKCC2, NHE1, and ENaC alpha- subunit increased significantly. On the contrary, the expression of AQP5 was significantly decreased, while that of AQP1 was not significantly altered. These findings indicate that the sodium transporters and water channels may be under a tonic regulatory influence of NO in the salivary gland.

Effects of Antioxidant Drugs in Rats with Acute Renal Injury

Acute renal failure is mainly caused by ischemia/reperfusion (I/R) injury or nephrotoxic drugs, in which reactive oxygen species (ROS) may play an important role. Therefore, antioxidants are expected to decrease the vulnerability of renal injury associated with oxidative challenges. alpha-Lipoic acid (alpha-LA), potent antioxidant, could act as ROS scavengers, iron chelators and enzyme modulators. In rats with acute renal injury, dysregulation of aquaporin (AQP) water channels and sodium transporters has been noted. I/R injury or cisplatin induced marked down-regulation of AQP1, AQP2 and AQP3 water channels, and type-3 Na-H exchanger, Na,K-ATPase, and Na-K-2Cl cotransporters, in association with impairment of urinary concentration and tubular sodium reabsorption. Treatment with alpha-LA prevented the dysregulation of AQP channels and sodium transporters, along with improved urinary concentrating capability and renal sodium reabsorption.

Sympathetic and parasympathetic regulation of sodium transporters and water channels in rat submandibular gland

The present study was aimed to explore the role of sympathetic and parasympathetic nerves in the regulation of sodium transporters and water channels in the salivary gland. Rats were denervated of their sympathetic and parasympathetic nerves to the submandibular gland, and the glandular expression of sodium transporters and water channels was determined by Western blot analysis. The expression of either alpha1 or beta1 subunit of Na, K-ATPase was not significantly affected either by the sympathetic or by the parasympathetic denervation. The expression of subunits of epithelial sodium channels was significantly increased both in the denervated and contralateral glands either by the sympathetic or by the parasympathetic denervation. Neither the sympathetic nor the parasympathetic denervation significantly altered the expression of aquaporin-1 (AQP1). Nor was the expression of AQP4 affected significantly by the parasympathetic or the sympathetic denervation. On the contrary, the expression of AQP5 was significantly increased not only by the parasympathetic but also by the sympathetic denervation. These results suggest that sympathetic and parasympathetic nerves have tonic regulatory effects on the regulation of certain sodium transporters and AQP water channels in the salivary gland.

Altered expression of sodium transporters and water channels following sympathetic and parasympathetic denervation in rat submandibular gland

The flow of saliva is controlled entirely by nervous stimuli. The present study was aimed to explore the role of sympathetic and parasympathetic nerves in the regulation of sodium transporters and water channels in the salivary gland. Rats were denervated of their sympathetic and parasympathetic nerves to the submandibular gland, and the expression of sodium transporters and water channels was determined. The expression of either alpha-1 or beta-1 subunit of Na, K-ATPase was not significantly affected by the sympathetic denervation. On the contrary, the expression of both subunits was decreased by the parasympathetic denervation. The expression of alpha-, beta-, and gamma-subunits of ENaC was not significantly affected by the sympathetic denervation, but was increased by the parasympathetic denervation. On the contrary, the expression of NHE3 was markedly decreased by both the sympathetic and the parasympathetic denervation. The sympathetic denervation significantly increased the expression of AQP1, while the parasympathetic denervation was without effect. The sympathetic and parasympathetic denervation significantly increased the expression of AQP4. The sympathetic denervation did not affect the expression of AQP5, but the parasympathetic denervation significantly decreased it. These results suggest that sympathetic and parasympathetic nerves have tonic effects on the regulation of sodium transporters and AQP water channels in the salivary gland. The sympathetic and parasympathetic denervation may then result in alterations of secretory rate and electrolyte composition of the saliva.

Pathogenesis of Postobstructive Diuresis: Role of Aquaporin Water Channels, Sodium Transporters and Natriuretic Peptide System in the Kidney in Rats

Although the obstruction is potentially reversible with treatment, marked and sometimes prolonged diuresis and natriuresis associated with an impaired ability to concentrate the urine may follow relief of the obstruction. Various factors contributing to the postobstructive diuresis and natriuresis have been suggested, including decreases of tubular sodium reabsorption, retention of urea and expansion of extracellular fluid volume. Tubular damage as a consequence of obstruction may occur one or more nephron segments and may result in decreased reabsorption of filtrate. The discovery of aquaporin (AQP) membrane water channels and sodium (co)transporters and channels provided insight, at the molecular level, into the fundamental physiology and pathophysiology of water and sodium balance. In addition, recent studies have shown that the kidney per se is also a site of production and release of atrial natriuretic peptide (ANP). The locally synthesized ANP may act in a paracrine manner to increase the urinary excretion of sodium and water. In this context, an altered regulation of ANP in the kidney may result in an altered urinary excretion. The combined interactions of multiple independent mechanisms are thought to be involved in the pathogenesis of postobstructive diuresis and natriuresis. We examined the changes of AQP water channels, sodium (co)transporters and natriuretic peptide system in obstructed kidneys. The expression of AQP water channels and sodium transporters was decreased in the obstructed kidneys, which may at least in part account for the urinary concentration defect associated with postobstructive diuresis and natriuresis. In addition, the postobstructive natriuresis was associated with an enhanced renal expression of ANP mRNA and an increased urinary excretion of ANP. The plasma dendroaspis natriuretic peptide (DNP) level was increased following an experimental ureteral obstruction. The urinary excretion of DNP was increased along with the postobstructive diuresis. An enhanced activity of DNP system may in part play a role in mediating the postobstructive diuresis