Effect of extracellular volume expansion on erythrocyte sodium transport in rats.

The effects of extracellular volume expansion (EVE) on the major sodium transport systems and sodium and potassium contents in rat erythrocytes have been examined in the present study. Study has been performed in anesthetized Wistar rat weighing about 300 g. Acute extracellular volume expansion (EVE) was induced by a constant intravenous saline infusion (3% body wt, 3 hours). Rats anaesthetized and catheterized but not expanded were used as controls. Arterial blood samples from control and expanded rats were obtained at the same time, and assayed immediately. Intracellular sodium and potassium concentration and ouabain sensitive (Na(+)-K(+)-pump) and bumetanide sensitive (Na(+)-K(+)-cotransport system) outward Na+ fluxes in erythrocytes were measured. The effect of plasma on erythrocyte transport was also analyzed by measuring 86Rb uptake. Neither of two plasma cations (Na+ and K+) were modified by the EVE. Also intracellular Na+ and K+ levels remained unvariable. Total Na+ efflux was not modified by EVE, but pump-mediated Na+ efflux was smaller after than before EVE. The ouabain-inhibible Na+ efflux rate constant decreased after EVE (from 687 +/- 81 to 525 +/- 29 h-1 x 10(-3); P less than 0.05). Both Na(+)-K(+)cotransport-mediated Na+ efflux and passive permeability increased significantly after EVE. The incubation with plasma from saline-infused animals induced a significant decrease in Rb uptake rate constant, that was not observed after incubation with plasma from non-expanded rats.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of solution non-ideality on membrane transport in three-dimensional models of the renal concentrating mechanism

Previous models of the renal concentrating mechanism employ ideal approximations of solution thermodynamics for membrane transport calculation. In three-dimensional models of the renal medulla, predicted urine concentrations reach levels where there idealized approximations begin to break down. In this paper we derive equations that govern membrane transport for non-dilute solutions and use these equations in a three-dimensional model of the concentrating mechanism. New numerical methods were employed that are more stable than those employed previously. Compared to ideal solution models, the urea non-ideality tends to increase predicted osmolarities, whereas NaCl non-ideality decreases predictions.     

Effect of amiloride on cell volume regulation in renal straight proximal tubules

Amiloride has been shown to impair cell volume regulatory decrease in amphiuma red cells. The present study has been performed to test for the influence of amiloride on volume regulatory decrease and electrical properties in isolated perfused mouse straight proximal tubules. Replacement of 40 mmol/l NaCl with 80 mmol/l mannitol in bath perfusate does not appreciably affect the cell volume or the potential difference across the basolateral cell membrane. Reduction of osmolarity by omission of mannitol leads to cell swelling by 16.7 +/- 0.7% (n = 7), followed by volume regulatory decrease to 107.2 +/- 1.2% (n = 7) of original cell volume within 2 min. 1 mmol/l amiloride (but not 0.1 mmol/l amiloride) in the bath depolarizes the basolateral cell membrane from -63 +/- 1 mV (n = 24) by +16 +/- 1 mV (n = 16), decreases the apparent potassium transference number from 0.69 +/- 0.02 (n = 5) to 0.36 +/- 0.05 (n = 5), and significantly impairs volume regulatory decrease without appreciably modifying cell volume in isotonic solutions. 1 mmol/l amiloride in the luminal perfusate leads to a slight hyperpolarization of the basolateral cell membrane but does not interfere with volume regulatory decrease. Reduction of bath osmolarity depolarizes the basolateral cell membrane within 30 s by +7.8 +/- 0.8 mV (n = 18) in the absence and by +18 +/- 2 mV (n = 8) in the presence of amiloride. In the presence of reduced bath osmolarity and amiloride the potassium transference number amounts to 0.36 +/- 0.04 (n = 8). The hyperpolarization following luminal application of amiloride is most likely due to inhibition of luminal sodium channels, whereas bath amiloride depolarizes the basolateral cell membrane by reduction of basolateral potassium selectivity. As in amphiuma red cells amiloride impairs volume regulatory decrease in proximal straight renal tubules.     

Effect of potassium on cell volume regulation in renal straight proximal tubules

Summary The present study was designed to assess for the influence of extracellular potassium and of inhibitors of potassium transport on cell volume regulatory decrease in isolated perfused straight proximal tubules of the mouse kidney. Volume regulatory decrease is virtually unaffected when bath potassium concentration is elevated from 5 to 20 mmol/liter, and still persists, albeit significantly retarded, in the presence of the potassium channel blocker barium on both sides of the epithelium and during virtually complete dissipation of the transmembrane potassium gradient by increasing extracellular potassium concentration to 40 mmol/liter. As evident from electrophysiologic observations, barium blocks the potassium conductance of the basolateral cell membrane. Reduction of bicarbonate concentration and increase of H⁺ concentration in the bath solution cannot compensate for enhanced potassium concentration and cell volume regulatory decrease is not affected in the presence of the K/H exchange inhibitor omeprazole. Similarly cell volume regulatory decrease is not affected by ouabain. In conclusion, potassium movements through potassium channels in the basolateral cell membrane are important determinants of cell volume and may participate in cell volume regulatory decrease. However, a powerful component of cell volume regulatory decrease in straight proximal tubules of the mouse kidney is apparently independent of potassium conductive pathways, K/H exchange and Na⁺/K⁺-ATPase.     

Hepatocytes volume regulation

Cell volume regulation has been studied during isolated dog liver perfusion. In presence of ouabain (10(-4) M) rapid but quantitatively matched exchange of K for Na occurs and the cellular volume is maintained until (90 min later) intracellular K concentration falls below 80 mEq/litre. Additional mechanism of protection of cell volume as loss of intracellular anions should also play a r?le since ouabain produces rapidly a membrane depolarization and chloride gain. A similar sequence of events is obtained when inhibition of the sodium pump is produced by anoxia but in this case the chloride gain in excess of cation gain is particularly marked. Submitted to an hypotonic shock the hepatocytes swell but tend to partially recover their volume by loosing K, indeed when osmolarity is corrected the cells maintain a sub-normal volume. Ouabain inhibits (or masks?) this iso-osmotic regulation. When submitted to an hypertonic medium a reduced cell volume is obtained and maintained for hours even in presence of ouabain, which produces a Na/K exchange at the same rate as in normal conditions.     

Effect of exogenous antioxidants on erythrocyte redox status and hepcidin content in disorders of iron metabolism regulation

In many diseases associated with impairments in iron metabolism, erythrocytes exhibit an increased sensitivity to oxidative stress induced in vitro. In this study we have examined the antioxidant status of erythrocytes from healthy donors and from 12 patients with disorders of iron homeostasis by measuring the extent of hemolysis induced in vitro by tert-butyl hydroperoxide (t-BHP). The extent of hemolysis observed with patient erythrocytes was significantly higher than that observed in experiments with erythrocytes from healthy donors. After therapeutic infusions of the antioxidants mexidol or emoxypin, oxidative hemolysis in patients was restored to normal values and blood hepcidin increased significantly as compared with its initial level. A significant correlation was observed between hepcidin concentration after treatment and t-BHP-induced hemolysis before treatment. These data suggest that antioxidants may exert a favorable effect on those at risk for iron overload disease.     

Mathematical analysis of human red blood cell volume regulation with regard to the elastic effect of the erythrocyte shell on metabolic processes

The mathematical model of the regulation of ion exchange and human erythrocyte volume is extended with a biomechanical model of the erythrocyte shell. This model was used to analyze the influence of elastic properties of the erythrocyte shell on erythrocyte volume in the experiments, where the volume of erythrocytes increased due to the formation of ion channels in the membrane after the treatment with amphotericin B and in case of placing red blood cells in a hypo-osmotic medium. During red blood cell deformation at a constant surface area up to sphericity, the influence of mechanical properties of the shell on volume regulation was shown to be negligible compared to the influence of ion exchange. Further osmotic swelling of red blood cells followed by the increase in their surface area is determined by tensile stiffness of the shell. The high value of tensile stiffness inherent to the erythrocyte shell is constraint for its volume change and also affects ion exchange.     

Principles of cell volume regulation

Cell volume is determined by the content of osmotically active solute (cell osmoles) and the osmolarity of the extracellular fluid. Cell osmoles consist of non-diffusible and diffusible solutes. A large fraction of the diffusible cation content balances negative charges on the non-diffusible solutes. The content of diffusible solutes is determined by the electrochemical gradients driving them across the plasma membrane and the availability and activity of transport pathways in the membrane. The classical view that the sodium pump offsets passive leaks must be modified to accommodate the contributions of a number of secondary active transport processes, as well as to allow for changes in cell nondiffusible osmoles and in their net negative charge. The behaviour of cells in anisosmotic media is often different from that predicted for a perfect osmometer. In many cases this is a consequence of changes in cell osmole content. However, caution is required in extrapolating from in vitro responses of isolated cells to large, acutely induced changes in medium osmolality to the responses of tissues in vivo to more subtle changes in extracellular osmolality.