Magali de Araujo

Angiotensin-(1-7) stimulates water transport in rat inner medullary collecting duct: evidence for involvement of vasopressin V2 receptors.

The peptide angiotensin-(1–7) [Ang-(1–7)] is known to enhance water transport in rat inner medullary collecting duct (IMCD). The aim of this study was to determine the mechanism of the Ang-(1–7) effect on osmotic water permeability (Pf). Pf was measured in the normal rat IMCD perfused in vitro in presence of agonists [Ang-(1–7), arginine vasopressin (AVP) and Ang-(3–8)], and antagonists of the angiotensin and the vasopressin cascade. Ang-(1–7), but not Ang-(3–8), increased Pf significantly. The effect of Ang-(1–7) on Pf was abolished by its selective antagonist, A-779, added before or after Ang-(1–7). Prostaglandin E2 and the protein kinase A inhibitor H8 also blocked the Ang-(1–7) effect. Blockade of vasopressin V1 receptors by antagonists did not change the Ang-(1–7) effect, but pre-treatment with a V2 antagonist abolished the effect of Ang-(1–7) on Pf. Similarly, pre-treatment with A-779 inhibited AVP’s effect on Pf. Forskolin-stimulated Pf was blocked both by A-779 and by the V2 antagonist. Finally, Ang-(1–7) increased cAMP levels in fresh IMCD cell suspensions whilst the forskolin-stimulated cAMP synthesis was decreased by A-779 and the V2 antagonist. These data provide evidence that Ang-(1–7) interacts via its receptor with the AVP V2 system through a mechanism involving adenylate-cyclase activation.

Magnesium Supplementation Combined with N-Acetylcysteine Protects against Postischemic Acute Renal Failure

Magnesium is a potent vasodilator whose effects have not been evaluated in renal ischemia. The antioxidant properties of N-acetylcysteine (NAC) partially protect animals from ischemic/reperfusion injury. This study was designed to evaluate magnesium supplementation, alone or combined with NAC, on ischemic acute renal failure. Rats were maintained on normal diets, supplemented or not with MgCl(2).6H(2)O (1% in drinking water) for 23 d, and some rats received NAC (440 mg/kg body wt) on days 20 to 23. On day 21, ischemia was induced by clamping both renal arteries for 30 min. Five groups were studied: Normal, ischemia, ischemia+magnesium, ischemia+NAC, and ischemia+magnesium+NAC. GFR (inulin clearance), renal blood flow (RBF), FEH(2)O, and FENa were determined. Serum magnesium was decreased in ischemia-only rats. Magnesium prevented postischemia GFR and RBF decreases but did not protect against tubular damage. However, NAC completely restored the tubular damage induced by ischemia/reperfusion. Semiquantitative immunoblotting showed that NAC prevented the decreased expression of Na-K-2Cl co-transporter and aquaporin 2 after renal ischemia/reperfusion. Untreated rats with acute renal failure demonstrated markedly decreased endothelial nitric oxide synthase expression. Significantly, treatment with NAC, magnesium, or both completely inhibited downregulation of endothelial nitric oxide synthase. The tubular necrosis scores were lower in rats that were treated with NAC alone or with the magnesium-NAC combination. Magnesium prevented postischemia GFR and RBF decreases but did not protect against tubular damage. The NAC protected tubules from ischemia, decreased infiltrating macrophages/lymphocytes, and had a mild protective effect on GFR. In ischemic/reperfusion injury, renal function benefits more from the magnesium-NAC combination than from magnesium alone.

Cyclooxygenase 2 inhibition suppresses tubuloglomerular feedback: roles of thromboxane receptors and nitric oxide

Thromboxane (TxA(2)) and nitric oxide (NO) are potent vasoactive autocoids that modulate tubuloglomerular feedback (TGF). Each is produced in the macula densa (MD) by cyclooxygenase-2 (COX-2) and neuronal nitric oxide synthase (nNOS), respectively. Both enzymes are similarly regulated in the MD and their interaction may be an important factor in the regulation of TGF and glomerular filtration rate. We tested the hypothesis that TGF is modified by the balance between MD nNOS-dependent NO and MD COX-2-dependent TxA(2). We measured maximal TGF during perfusion of the loop of Henle (LH) by continuous recording of the proximal tubule stopped flow pressure response to LH perfusion of artificial tubular fluid (ATF) at 0 and 40 nl/min. The response to inhibitors of COX-1 (SC-560), COX-2 [parecoxib (Pxb)], and nNOS (l-NPA) added to the ATF solution was measured in separate nephrons. COX-2 inhibition with Pxb reduced TGF by 46% (ATF + vehicle vs. ATF + Pxb), whereas COX-1 inhibition with SC-560 reduced TGF by only 23%. Pretreatment with intravenous infusion of SQ-29,548, a selective thromboxone/PGH(2) receptor (TPR) antagonist, blocked all of the SC-560 effect on TGF, suggesting that this effect was due to activation of TPR. However, SQ-29,548 only partially diminished the effect of Pxb (-66%). Specific inhibition of nNOS with l-NPA increased TGF, as expected. However, the ability of Pxb to reduce TGF was significantly impaired with comicroperfusion of l-NPA. These data suggest that COX-2 modulates TGF by two proconstrictive actions: generation of TxA(2) acting on TPR and by simultaneous reduction of NO.

Tubuloglomerular feedback is decreased in COX-1 knockout mice after chronic angiotensin II infusion

Prostaglandins (PGs), produced by two isoforms of cyclooxygenase (COX), COX-1 and COX-2, are important modulators of renal hemodynamics. COX-1 and COX-2 are expressed in the kidney often at distinct sites. Thromboxane (TxA(2)), PGE(2), and prostacyclin (PGI(2)) are the major PGs in the renal cortex of mice. Acute infusion of the vasoconstrictor ANG II increases COX-2-dependent PGE(2) and PGI(2). COX-2 is primarily expressed in the macula densa (MD), where several PG synthases are also expressed. We previously showed that MD COX-2 products modulate tubuloglomerular feedback (TGF) in the rat. Genetic deletion of COX-1 enhances COX-2 production of PGs, decreases renal and urinary PGs, and attenuates ANG II-induced hypertension. The present study tested the effects of chronic ANG II infusion on TGF in COX-1 knockout (KO) mice. Basal TGF was similar in COX-1 KO and wild-type (WT) mice. Chronic ANG II infusion increased TGF in WT mice (WT: 9.3 +/- 0.7 vs. WT + ANG II: 12.2 +/- 1.6 mmHg, P < 0.02). However, chronic ANG II decreased TGF in COX-1 KO mice (KO: 11.4 +/- 1.1 vs. KO + ANG II: 8.3 +/- 0.6 mmHg, P < 0.01). Pretreatment with the COX-2 inhibitor SC-58,236 in COX-1 KO mice prevented the ANG II-associated reduction in TGF (11.4 +/- 1.0 vs. 11.5 +/- 0.28 mmHg, not significant). Excretion of 6-keto-PGF(2alpha), the metabolite of PGI(2), was increased by ANG II infusion, whereas excretion of TxB(2), the stable metabolite of TxA(2), was not changed. ANG II infusion increased mean arterial pressure similarly in both WT and KO mice (WT: 93 +/- 2 vs. KO: 92 +/- 3 mmHg), but not in KO mice pretreated with SC-58,236 (85 +/- 2 mmHg). This study shows that COX-1-generated PGs partially mediate ANG II increases in TGF and that COX-2 PGs offset that effect.

Aquaporin 2 expression increased by glucagon in normal rat inner medullary collecting ducts

It is well known that Glucagon (Gl) is released after a high protein diet and participates in water excretion by the kidney, principally after a protein meal. To study this effect in in vitro perfused inner medullary collecting ducts (IMCD), the osmotic water permeability (Pf; mum/s) at 37 degrees C and pH 7.4 in normal rat IMCDs (n = 36) perfused with Ringer/HCO(3) was determined. Gl (10(-7) M) in absence of Vasopressin (AVP) enhanced the Pf from 4.38 +/- 1.40 to 11.16 +/- 1.44 microm/s (P < 0.01). Adding 10(-8), 10(-7), and 10(-6) M Gl, the Pf responded in a dose-dependent manner. The protein kinase A inhibitor H8 blocked the Gl effect. The specific Gl inhibitor, des-His(1)-[Glu(9)] glucagon (10(-7) M), blocked the Gl-stimulated Pf but not the AVP-stimulated Pf. There occurred a partial additional effect between Gl and AVP. The cAMP level was enhanced from the control 1.24 +/- 0.39 to 59.70 +/- 15.18 fm/mg prot after Gl 10(-7) M in an IMCD cell suspension. The immunoblotting studies indicated an increase in AQP2 protein abundance of 27% (cont 100.0 +/- 3.9 vs. Gl 127.53; P = 0.0035) in membrane fractions extracted from IMCD tubule suspension, incubated with 10(-6) M Gl. Our data showed that 1) Gl increased water absorption in a dose-dependent manner; 2) the anti-Gl blocked the action of Gl but not the action of AVP; 3) Gl stimulated the cAMP generation; 4) Gl increased the AQP2 water channel protein expression, leading us to conclude that Gl controls water absorption by utilizing a Gl receptor, rather than a AVP receptor, increasing the AQP2 protein expression.

Effect of Low and High Potassium Diets on H+-K+-ATPase and Na+-K+-ATPase Activities in the Rat Inner Medullary Collecting Duct Cells

This study was designed to measure H-K-ATPase and Na-K-ATPase activities in rat inner medullary collecting duct (IMCD) cells during potassium depletion and loading. Both enzyme activities were similarly distributed from the proximal to the distal portion of this nephron segment. One week K depletion did not stimulate H-K-ATPase activity but reduced Na-K-ATPase activity. Within 2 weeks after the onset of potassium depletion, the rats developed metabolic alkalosis, and H-K-ATPase activity was suppressed while Na-K-ATPase activity had returned to control values. H-K-ATPase activity was suppressed following potassium loading whereas Na-K-ATPase activity was unchanged. The results are consistent with the presence of H-K-ATPase in IMCD and indicate that H-K-ATPase and Na-K-ATPase activities are modulated by potassium intake.

Study of Erythrocyte ATPases in Infants Evaluated during the Recovery Phase of Severe Dehydration Caused by Diarrhea

Background/Aims: Patients severely dehydrated from diarrhea are at risk of developing hyperkalemia consequent to fluid therapy treatment. In parallel with the regulation of external potassium balance by the kidney and gastrointestinal tract, plasma potassium is rapidly regulated by redistribution of potassium between the extracellular and intracellular compartments. Erythrocytes contain ATPases that play a role in this potassium movement. In this study, erythrocyte ATPase effectiveness was evaluated in infants dehydrated from diarrhea and compared to that of healthy infants. Methods: Blood samples were collected from dehydrated and healthy infants. The activity of Na+,K+-ATPase and of an ouabain-insensitive K+-ATPase were assessed. Serum electrolytes and blood pH were also determined. Results: No hyperkalemia was found, even in dehydrated infants presenting with severe hyperchloremic metabolic acidosis. In the erythrocytes of dehydrated infants, Na+,K+-ATPase activity was increased correlating positively with the amount of sodium administered. High K+-ATPase activity in the erythrocytes correlated with low plasma potassium. The K+-ATPase activity correlated positively with the amount of potassium administered to dehydrated infants. Conclusion: These findings suggest that the erythrocytes Na+,K+-ATPase and K+-ATPase both protect against plasma potassium abnormalities in dehydrated infants. In such infants, the risk of hyperkalemia is probably low.

Erratum by the Publisher – Announcement

In continuation of our tradition of joint congresses, the meeting in Feiburg will focus on new experimental and clinical data. The program, consisting of posters, free contributions, and invited reviews, will have three focuses: (1) basic research; (2) experimental clinical science, and (3) lectures on hot clinical topics. The congress also encompasses free symposia, plenary sessions, and a social program. The major topics will be defined by the Joint Program Committee of the Gesellschaft für Nephrologie and the Deutsche Arbeitsgemeinschaft für Klinische Nephrologie. The congress language will be German. Free communications or posters should be submitted by using the specific abstract form which, together with the preliminary program, will be available in February 1999. The success of the meeting will depend firstly on broad participation of nephrologists and those interested in nephrology and secondly on the contributions submitted. We will try to generate an interesting program and secure sufficient time also for social activities.

ACTIVITY OF Na, K-ATPase DURING RECOVERY OF INFANTILE DIARRHEAL DEHYDRATION 1

Severe electrolyte disorders occur frequently associated with diarrheal dehydration in infants. With this objective we evaluated the erythrocyte activity of Na,K-ATPase in 9 infants dehydrated (D) by diarrhea, on the first and seventh day of recovery. A control group, constituted by 5 normal infants(CI) in addition to another group of 6 normal children (2 to 6 years old-CC) were compared. Na,K-ATPase activity of D was elevated in the first(540±64) and in the seventh day (435±45 nMPi/mg p/h) compared with controls (220±22nMPi/mgp/h). In the D, hyperchloremic metabolic acidosis was associated with normal serum Na and K concentrations. In controls, Na,K-ATPase activity increased with age (r=0,73, p<0,01) and showed an inverse correlation with serum K (r=0,65, p<0,01). Plasma aldosterone concentration was significantly lower in CC(18±4,5,p<0,01) than Cl (73 ± 13 ng/dl). There was an inverse linear correlation between (Na) y and Na,K-ATPase activity (r=0,73; p<0,01) and a positive correlation between the amount of Na administered and Na,K-ATPase activity (r=0,93; p<0,01): In conclusions: 1) The maturity of Na,K-ATPase was related to extra renal regulation of K; 2) the lower activity of this group of this pump in infants is not due to aldosterone deficiency; 3) Na,K-ATPase of crythrocytes was activated maintaining an important electrolyte regulation. This activation is probably a consequence to the K deppletion that occurs in diarrhea.