Alkali Therapy Attenuates the Progression of Kidney Injury via Na/H Exchanger Inhibition in 5/6 Nephrectomized Rats

Metabolic acidosis is a cause of renal disease progression, and alkali therapy ameliorates its progression. However, there are few reports on the role of renal acid-base transporters during alkali therapy. We evaluated the effect of sodium bicarbonate therapy and the role of acid-base transporters on renal disease progression in rats with a remnant kidney. Sprague-Dawley rats consumed dietary sodium bicarbonate (NaHCO3) or sodium chloride (NaCl) with 20% casein after a 5/6 nephrectomy. After being provided with a casein diet, the NaHCO3-treated group had higher levels of serum bicarbonate than the control group. At week 4, the glomerular filtration rate in the NaHCO3 group was higher than that in the NaCl group, and the difference became prominent at week 10. The glomerulosclerosis and tubulointerstitial damage indices in the NaHCO3 group were less severe compared with controls at week 4 and 10. The expression of the Na/H exchanger (NHE) was decreased, and apical reactivity was decreased in the NaHCO3 group, compared with the NaCl group. Endothelin-1 levels in the kidney were also decreased in the NaHCO3 group. Dietary sodium bicarbonate has the effects of ameliorating renal disease progression, which may be related to the altered expression of NHE in the remaining kidney.

Modulación inflamatoria en el shock traumático: [revisión] / Inflammatory modulation in traumatic shock: [review]

Hemorrhagic hypovolemic shock secondary to trauma is an important cause of morbidity and mortality worldwide. During the last few years, new concepts have emerged and the guidelines of fluid resuscitation in these patients have been redefined. The concept of hypotensive resuscitation has been established and new colloid solutions based on starch have been manufactured, been hydroxyethyl starch in a balanced electrolytic solution, the most studied and successful one. It has been reported, as well, the positive effects of the pharmacologic modulation of the inflammatory pathways in experimental model subjects submitted to hypovolemic shock. Products such as, ethyl pyruvate and the Na+/H+ type 1 inhibitor, BIIB513, have been Studies only experimentally in rodent models using colloids as the primary resuscitation fluid. The significant improvement in the hemodinamyc, pattern and the cardiac and inflammatory indexes and mediators, has created the basis for their use in clinical trials in the near future. The systemic inflammatory response is an important cause of multiple organ failure that increases the late mortality of patients surviving the initial early phases of hypovolemic traumatic shock and its experimental modulation in rodent models with products such as ethyl pyruvate and BIIB513 has produced excellent in vivo and in vitro results.

Universalmente se considera el Shock hipovolémico de origen hemorrágico como una importante causa de morbi-mortalidad. Durante los últimos años se ha redefinido los conceptos de la reanimación con líquidos intravenosos en los pacientes con choque hipovolémico y establecido los conceptos de reanimación hipotensa con el uso de nuevos coloides derivados del almidón, tales como el hidroxietil-almidón en solución electrolítica balanceada (Hextend®). Así mismo, se ha reportado el beneficio que conlleva el uso de modificadores de la cascada inflamatoria en modelos experimentales de sujetos sometidos a choque hipovolémico hemorrágico. Productos como el etil piruvato y la BIIB513, un inhibidor selectivo del intercambiador Na+/H+ tipo 1, han sido estudiados sólo experimentalmente en modelos roedores, empleando coloides como principal elemento de reanimación. Al mejorar el perfil hemodinámico, parámetros cardíacos y niveles de mediadores inflamatorios, estos compuestos constituyen una base cierta para ser incluidos en estudios clínicos en un futuro próximo. La respuesta inflamatoria sistémica está íntimamente implicada en la patogénesis de la Falla Orgánica Múltiple, aumentando la mortalidad tardía de pacientes que sobreviven las etapas tempranas del shock hipovolémico hemorrágico traumático. Su modulación experimental con el etil piruvato o bien la BIIB513 ha dado excelente resultado tanto en modelos experimentales in vivo como in vitro.

Myocardial protection by amiloride, a Na[+]-H[+] exchange blocker, after in vivo resuscitation of hemorrhagic shock in rats

Hemorrhagic shock and resuscitation is known to result in moyocardial contractile dysfunction and failure, but the role of the Na[+]-H[+] exchanger in hemorrhagic shock remains unclear. However, there has been intensive research investigating the myocardial protective effects on the Na[+]-H[+] exchange blockers in ischemia-reperfusion. Prior studies in our laboratory found that blocking the Na[+]-H[+] exchanger for short period on an ex vivo perfusion of isolated hearts has a protective effect on myocardial contractile function. The present study examined the protective effects of blocking the cardiac Na[+]-H[+] exchanger, by injecting 100 microM amiloride intra-arterially, on myocardial contractile function after in vivo resuscitation following our hour of hemorrhagic shock in rats. Male Sraque-Dawley rats were assigned to either hemorrhagic-treated, untreated, or similar time-matched control groups [n= 4 per group]. The left carotid artery was cannulated for blood pressure monitoring and hemorrhaging. Rats were hemorrhaged using a reservoir model. Arterial blood pressure was maintained at a mean arterial blood pressure of 40 mm Hg. After a 60-min hemorrhagic shock, rats were treated or not by injection of 1 ml of 100 microM amiloride [0.027 mg/ml] intra-arterially. Rats were then resuscitated in vivo and monitored for 30 min. Hearts were harvested and perfused in the Langendorff system for 60 min for measurement of hemodynamic function. Inhibition of the Na[+]-H[+] exchanger by injecting amiloride i.a. before in vivo resuscitation following hemorrhagic shock improved myocardial contractile function. Blocking the Na[+]-H[+] exchanger using amiloride before in vivo resuscitation following hemorrhagic shock protected the heart against dysfunction

Myocardial protection by Ethyl-isopropyl amiloride, a specific NA[+] -H[+] exchange inhibitor, following hemorrhagic shock

Hemorrhagic shock and resuscitation is well known to result in myocardial dysfunction and injury. Stimulation of the Na[+] -H[+] exchanger plays an important role in the pathway of myocardial injury. The purpose of the present study was to examine the protective effects of blocking the cardiac Na[+] -H[+] exchange, using 100mM ethyl-isopropyl amiloride [EIPA], a specific Na[+] -H[+] exchanger blocker, on myocardial contractile function on ex vivo resuscitation of isolated rat heart following one hour of hemorrhagic shock. Sprague- Dawley rats were assigned to hemorrhage, hemorrhage + EIPA, sham hemorrhage and sham hemorrhage + EIPA groups. Rats were hemorrhaged for one hour. Hearts were harvested and ex vivo treated and resuscitated by perfused in the Langendorff System. Myocardial function was determined. The results showed that inhibition of the Na[+] -H[+] exchanger using EIPA improved the post-resuscitation myocardial contractile function. Blocking the Na[+] -H[+] exchanger using 100mM EIPA following 60 minutes of hemorrhagic shock improved myocardial function

Analisis ecocardiografico del efecto de diferentes inhibidores del intercambiador Na+/H+ sobre la estructura y funcion sistolica del ventriculo izquierdo en ratas espontaneamente hipertensas / Echocardiographic analysis of the effect of different Na+/H+ exchanger inhibitors on left

El presente estudio fue proyectado para analizar mediante ecocardiograma los efectos del HOE 642 (cariporide) (HOE) y del BIIB 723 (BIIB) sobre la estructura y función sistólica del ventrículo izquierdo en ratas espontáneamente hipertensas (SHR)- 8 con 30 mg/kg/día de HOE, 8 con 30 mg/kg/día de BIIB durante 30 días y 4 sin tratamiento (grupo control) durante esos 30 días. Los distintos parámetros analizados no mostraron cambios durante ese período en las ratas controles. Si bien el HOE determinó un leve descenso de la presión arterial (C: 184 ± 1.75 mm Hg; 30d:176.20 ± 2.60 mm Hg, p <0.01), no detectada con BIIB,ambas drogas provocaron un aumento del estrés sistólico pico (HOE C:166 ± 29 kdinas/cm2; 30d: 204 ± 34kdinas/cm2, p <0.04. BIIB C: 164 ± 25.90 kdinas/cm2; 30d: 234 ± 29.30 kdinas/cm2, p<0.02).Tanto HOE comoBIIB redujeron significativamente la masa ventricular izquierda (MVI) (HOE C: 612.50±50 mg; 30d:452 ± 37 mg,p <0.01; BIIB C: 544 ± 16mg; 30 d: 374 ± 25 mg, p<0.01). El porcentaje de acortamiento endocárdico no semodificó luego del tratamiento con HOE (C: 62.30 ± 2.75; 30d 65.50 ± 2.40%, ns) y BIIB (C: 63.20 ± 2.39%;30d 67.20 ± 1.62%, ns). Los resultados analizados permiten concluir que ambos inhibidores determinaron similarreducción de la MVI. Esa reducción se acompañó de mejoría en los índices de evaluación de la función sistólica ventricular izquierda, pese al incremento del estrés sistólico pico. Estas evidencias sugieren que independientemente del inhibidor utilizado se encuentra un aumento del inotropismo, previamente comprometidodurante el desarrollo de la hipertrofia

The aim of this study was to analyze by echocardiogram, the action of two Na+/H+ exchange, inhibitors, HOE 642 (HOE) and BIIB 723 (BIIB) on left ventricular (LV) mass and LV systolic function. We studied 16 spontaneously hypertensive rats (SHR), 8 treated with HOE 30 mg/kg/day, 8 with 30 mg/kg/day of BIIB during 30 days and 4 SHR as controls during those 30 days. Results are expressed as mean values ± SEM. The systolic blood pressure and theechocardiograpic parameters examined did not evidence changes during that period in the controls rats. Eventhough HOE determined a slight decrease in blood pressure (HOE C: 184 ± 1.75 mm Hg; HOE 30d: 176.20 ±2.60 mm Hg - p <0.01) which was not detected with BIIB, both drugs provoked an increase of peak systolic stress (HOE C: 166 ± 29 kdynes/cm2; HOE 30d: 204 ± 34 kdynes/cm2, p <0.04; BIIB C: 164 ± 25.90 kdynes/cm2; BIIB 30d: 234 ± 29.30 kdynes/cm2, p <0.02). HOE and BIIB reduced LV mass after 30 days of administration (HOE C: 612.50 ± 50 mg; 30d: 452 ± 37 mg, p <0.01. BIIB C: 544 ± 16mg; 30d: 374 ± 25 mg, p <0.01). LV endocardial shortening was similar independently of the NHE inhibitors used (HOE C: 62.30 ± 2.75%; 30d: 65.50 ± 2.40%, ns. BIIB C: 63.20 ± 2,39%; 30d 67,20 ± 1.62%, ns). These data demonstrate that long-treatment with HOE or BIIB produced similar LV mass regression without changes in endocardial fractional shortening in spite of the increase of peak systolic stress. This finding could represent an increased inotropism previously depressed by the development of hypertrophy

Effects of cardiac Na[+]- H[+] exchange blockade on myocardial contractile function during hemorrhagic shock

Myocardial contractile dysfunction has been described following hemorrhagic shock. While the Na [+]-H[+] exchanger is well known to be a major regulator of intracellular pH. its role in hemorrhagic shock remains unclear. However, there has been intensive research investigating the myocardial protective role of several Na[+]-H[+] exchangers in ischemia-reperfusion, which is similar to hemorrhagic shock as both result in extracellular acidosis. The purpose of the present study was to examine the effects of blocking the cardiac Na[+]-H[+] exchanger, using 100 micro M. amiloride, on myocardial contractile function after ex vivo perfusion of isolated rat heart following one hour of hemorrhagic shock. Anesthetized male Sprague-Dawley rats were assigned to either hemorrhagic treated or untreated groups, or a similar time-matched control group [n=6 per group]. Rats were hemorrhaged using a reservoir model. Arterial blood pressure was monitored for one hour, and maintained at a mean arterial blood pressure of 40 mm Hg, via an intra-arterial catheter inserted into the left carotid artery. Two arterial blood samples were taken, one at baseline and another at 60 minutes of hemorrhage. These blood samples were analyzed for pH and lactate levels. Hearts were harvested and perfused either with a balanced salt solution for 60 minutes or a solution containing 100 micro M amiloride. Myocardial function was determined with a balloon- tipped catheter inserted into the left ventricle via the mitral valve. Indices of left ventricular function were measured, including left ventricular end diastolic pressure [LVEDP], left ventricular peak systolic pressure [LVPSP], coronary perfusion pressure [Pp] and left ventricular balloon volume [BV]. The left ventricular +/- dP/dt, which is the left ventricular index of contractility, was calculated. Left ventricular compliance [C] was also calculated. The results showed that inhibition of the Na[+]-H[+] exchanger for 5 minutes of ex vivo perfusion of the isolated hearts following hemorrhagic shock, improved myocardial contractile function as compared to the hemorrhage untreated hearts. The results also showed that hemorrhagic shock decreased myocardial contractile function as compared to controls. Blocking the Na[+]-H[+] exchanger for a short period on ex vivo perfusion of isolated hearts has a protective effect on myocardial contractile function. Further research is needed to investigate the exact mechanism of protection using more specific Na[+]-H[+] exchanger inhibitors

El intercambiador Na+/Ca2+ como responsable del atontamiento miocardio / The Na+/Ca2+ exchanger as responsible of myocardial stunning

Nuestro objetivo fue examinar la participación de los intercambiadores Na+/H+ (NHE) y Na+/Ca+2 (NCX) sobre las alteraciones sistólicas propias del atontamiento miocárdio. Se utilizó el modelo de corazón aislado isovolúmico de rata el cual fue sometido a 20 minutos de isquemia global (Is) y 30 minutos de perfusión (R). Este protocolo se repitió en prasencia de 1microM de HOE 642, bloqueante específico del NHE-1 y de KB-R7943, bloqueante del modo inverso del NCX (entrando Ca+2 y sacando Na de la célula) administrados antes de la Is, y/o en la R. En los controles isquémicos la contractilidad, evaluada a través de la +dP/dtmax se recuperó aproximadamente un 60 porciento. La recuperación postisquémica fue total cuando el bloqueo de NHE fue efectuado antes de la Is o al comienzo de la R y mejoró significativamente cuando en Is y R bloqueó el NCX ( 95 + o - 7 porciento). La contractura isquémica disminuyó com ambos tratamientos cuando fueron realizados previos a la Is. El aumento de la PDF observado en la R (24 + o - 6 y 12 + o - 2 mmHg) y por el bloqueo del NCX realizado durante Is y R (12 + o - 6 mmHg). Estos resultados indican que la activación del modo inverso del NCX secundaria a una activación del NHE (que incrementa el Na+ intracelular) durante la isquemia y reperfusión es el mecanismo culpable de la sobrecarga de Ca+2 involucrado en la disminuición de la contractilidad que caracteriza al miocardio atontado.

Changes in the expression of c-myc, RB and tyrosine-phosphorylated proteins during proliferation of NIH 3T3 cells induced by hyaluronic acid

We have shown that hyaluronic acid stimulates the proliferation of quiescent NIH 3T3 cells. We have shown that treatment of 1 mg/ml hyaluronic acid results in increase of tyrosine phosphorylation of two proteins, MW 124 kDa and 60 kDa as detected by anti-tyrosine antibodies by Western blot analysis. Maximum phosphorylation occurred within 2 h after addition of 1 mg/ml hyaluronic acid. Stimulation of proliferation was also accompanied by increase in c-Myc protein, which was inhibited by amlloride, an inhibitor of Na+/H+ antiporter and EGTA and increase in the steady state level of pRb, the RB gene product. These results suggest that the intracellular signal transduction pathways that mediate the stimulatory effects of hyaluronic acid on cellular proliferation are similar to those of growth factors.

Blockade of Na+/H+ exchanger contracts the portal vein of spontaneously hypertensive and Wistar Kyoto rats.

It has been claimed that the activity of Na+/H+ exchanger (NHE) is altered in spontaneously hypertensive rats (SHR) suggesting a possible role for it in the pathophysiology of hypertension. The purpose of this study has been to investigate the effect of blockade of NHE on the noradrenaline (NA) and 26K+ induced tone and on the recovery of tone from acid induced contractions in the portan vein of spontaneously hypertensive rats (SHR) as compared to their controls of Wistar Kyoto rats (WKY). Blockade of NHE by 10(-4) dimethylamiloride (DMA) raised the tone of NA and 26K+ activated preparation of both strains, the contractions being higher with NA activation. Total blockade of NHE by replacement of Na in solution with N-methy-D-Glucamine (NMDG) raised the tone of the NA activated preparations by 45+/-10%, n=8, P<0.01 and 33+/-4%, n=12, P < 0.01 in SHR and WKY respectively. The time for 50% relaxation from NH4Cl washout contraction was significantly prolonged by 10(-5) and 10(-4) M DMA in both strains under NA or 26K+ activation. DMA effect was greater on NA than on 26K+ activated preparations, and was not significantly different when comparing SHR to WKY results. In conclusion the results reported in this study indicate that NHE has similar activity in WKY and SHR portal veins and that its blockade contracts both preparations. Therefore, it is unlikely that increased NHE activity, acting directly on vascular smooth muscle tone, could be a primary cause for hypertension.