Inhibition Of NaCl Absorption Research Articles

Nitric oxide-inhibited chloride transport in cortical thick ascending limbs is reversed by 8-iso-prostaglandin-F2α.

Sodium chloride (NaCl) reabsorption in the cortical thick ascending limb (cTAL) is regulated by opposing effects. Nitric oxide (NO) inhibits NaCl reabsorption while 8-iso-prostaglandin-F2α (8-iso-PGF2α) stimulates it. Their interaction has not been evaluated in the cTAL. Because 8-iso-PGF2α has considerable stability while NO is a free radical with a short half-life, we hypothesized that, in the cTAL, the inhibition of NaCl absorption will be reversed by 8-iso-PGF2α. Chloride absorption (JCl) was measured in isolated perfused cTALs and whether the activation of protein kinase A (PKA) is required for this interaction. Since cyclic adenosine monophosphate (cAMP) is a major messenger for the 8-iso-PGF2α signaling cascade, and NO inhibits JCl by decreasing cAMP bioavailability, we measured 8-iso-PGF2α-stimulated cAMP in the presence of sodium nitroprusside (SNP). The NO donor, SNP (10-6 M), decreased JCl by 41%, while luminal 8-iso-PGF2α (100 μM) increased JCl to 315 ± 46 pmol/ min/mm (p < 0.003), reversing the effects of the NO donor. SNP inhibited JCl, 8-iso-PGF2α failed to increase JCl in the presence of H89. Basal cAMP was 56 ± 13 fmol/min/mm, in the presence of SNP 57 ± 6 fmol/min/mm, and 8-iso-PGF2α increased it to 92 ± 2 fmol/min/mm (p < 0.04). We concluded that 1) NO-induced inhibition of JCl in the cTAL can be reversed by 8-iso-PGF2α, 2) 8-iso-PGF2α and NO interaction requires PKA to control JCl, and 3) in the presence of NO, 8-iso-PGF2α continues to stimulate JCl because NO cannot reverse 8-iso-PGF2α-stimulated cAMP level.

P2X Receptor‐Mediated Inhibition Of NaCl Absorption In The Thick Ascending Limb: No Evidence For Nitric Oxide As Second Messenger

BackgroundBasolateral (bl.) P2X receptors acutely and markedly reduce NaCl absorption in mouse medullary thick ascending limb (mTAL). It has been proposed that P2X receptor mediate NO synthesis and subsequent NKCC2 inhibition. P2X receptor stimulation causes an increase in cytosolic Ca2+ and therefore this could be proximal to the generation of NO.ObjectiveHere we tested if blocking NO synthesis, adding an NO donor or hindering Ca2+ influx affects the ATP‐mediated (P2X) transport inhibition.MethodsWe used isolated, perfused mTALs from mice to electrically measure NaCl absorption. By microelectrodes we determined the transepithelial voltage (Vte) and the transepithelial resistance (Rte) and via these the transepithelial Na+ absorption (equivalent short circuit current, I′sc).ResultsWe confirm that bl. ATP (100μM) induced a marked, acute and reversible inhibition of Na+ absorption (28% ± 6%, n=6). In the presence of the NO synthase blocker L‐NAME (100 μM, ~4 min preincubation) the ATP effect remained unaffected (23% ± 9%, n=6). Also higher concentrations and longer incubation with L‐NAME (1 mM, 15min incubation) were without effect. High concentrations of L‐NAME (1 mM) per se reduced transport significantly. Acute addition of the NO donor SNAP (100μM, ~10 min) did not affect tubular transport. Furthermore, the removal of extracellular Ca2+ (100 nM, 2 min preincubation) had no effect on the ATP‐induced transport inhibition ([Ca2+ ]o=100nM: 21% ± 6%, n=5, [Ca2+]o=1μM: 24% ± 4%, n=6).ConclusionWe find no evidence for nitric oxide (NO) as second messenger for P2X receptor‐dependent transport inhibition. Similarly, Ca2+ signaling appears not involved in the ATP effect. It remains undefined how P2X receptors trigger the marked reduction of transport in the TAL.

Prostaglandin E2 regulation of ion transport is absent in medullary thick ascending limbs from SHR.

Regulation of HCO3- and Cl- absorption by arginine vasopressin (AVP) and prostaglandin E2 (PGE2) was examined in isolated, perfused medullary thick ascending limbs (MTAL) from 4- to 7-wk-old spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats. AVP inhibited HCO3- absorption by 50% at 10(-10) M and by 25% at 2 x 10(-12) M in MTAL from both WKY and SHR. Cholera toxin (10(-9) M) or forskolin (10(-6) M) in the bath also inhibited HCO3- absorption by 50% in the SHR. In MTAL from WKY, PGE2 (10(-6) M in the bath) increased HCO3- absorption from 7.1 +/- 0.4 to 12.0 +/- 0.4 pmol.min-1.mm-1 (P < 0.005) and decreased Cl- absorption from 65 +/- 7 to 47 +/- 6 pmol.min-1.mm-1 (P < 0.001) in the presence of 10(-10) M AVP. Under the same conditions, PGE2 had no effect on HCO3- or Cl- absorption in MTAL from SHR. PGE2 also reversed submaximal inhibition of HCO3- absorption by 2 x 10(-12) M AVP in WKY but not in SHR. With 10(-10) M AVP in the bath, phorbol 12-myristate 13-acetate (10(-6) M in the bath) increased HCO3- absorption from 6.6 +/- 0.5 to 12.3 +/- 0.4 pmol.min-1.mm-1 in MTAL from WKY and from 7.6 +/- 0.7 to 12.6 +/- 1.2 pmol.min-1.mm-1 in MTAL from SHR (P < 0.005). These results demonstrate that 1) the effects of PGE2 to stimulate HCO3- absorption and inhibit Cl- absorption in the presence of AVP are absent in MTAL from SHR, 2) the defect may involve an inability of PGE2 to stimulate protein kinase C, and 3) regulation of HCO3- absorption by AVP via adenosine 3',5'-cyclic monophosphate is similar in MTAL from WKY and SHR. The lack of PGE2 inhibition of NaCl absorption in the MTAL may contribute to renal salt retention during the development of hypertension in the SHR.

CGMP and Ca2+ regulation of ion transport across the isolated porcine distal colon epithelium

Intact epithelium from the porcine distal colon was stripped of serosal muscle and mounted in Ussing chambers to investigate the regulation of Na, Cl, and K transport by guanosine 3',5'-cyclic monophosphate (cGMP) and elevations in intracellular [Ca2+]. Under voltage-clamped conditions cGMP (250 microM) produced an increase in tissue short-circuit current (Isc) that reached a maximal value within 10-20 min and remained elevated > 40 min. This response was associated with an inhibition of NaCl absorption and stimulation of Cl and K secretion. In the absence of Cl the Isc also slowly increased but returned to baseline values within 20 min. Bicarbonate removal from both serosal and mucosal solutions or serosal bumetanide (20 microM) reduced the effect of cGMP on Isc by approximately 40%. When performed simultaneously, these conditions reduced the cGMP response by approximately 60%. Transepithelial Na and Cl flux measurements indicated that serosal bumetanide blocked increased Cl secretion without effecting changes in NaCl absorption. In contrast, mucosal amiloride blocked the effects of cGMP on NaCl absorption but not Cl secretion. The cGMP Isc response was potentiated in the presence of 1 mM, but not 10 microM, amiloride. Moreover, 1 mM amiloride inhibited Isc under control conditions but was ineffective in the presence of cGMP. The Ca2+ ionophore ionomycin (3 microM) produced a transient increase in the Isc that was also associated with a decrease in transepithelial NaCl absorption and an increase in Cl and K secretion. In contrast to cGMP, the ionomycin Isc response was eliminated after Cl removal from the bath.(ABSTRACT TRUNCATED AT 250 WORDS)

Brush-border tyrosine phosphorylation stimulates ileal neutral NaCl absorption and brush-border Na(+)-H+ exchange.

The drug genistein, a tyrosine (Tyr) kinase inhibitor, was used to define a role for Tyr phosphorylation in regulation of basal and stimulated neutral NaCl absorption in rabbit ileum. Brush-border vesicles contain Tyr-phosphorylated peptides. Genistein freeze-thawed into the vesicles caused a concentration-dependent inhibition of at least three peptides with M(r) 111,000, 83,000, and 80,000. Studied with the Ussing chamber-voltage clamp technique, genistein added to the ileal mucosal surface inhibited neutral NaCl absorption. Direct addition of genistein to brush-border vesicles made from ileal villus cells inhibited brush-border Na(+)-H+ exchange but not D-glucose-stimulated Na+ uptake. These effects were not duplicated by genistin, a drug with similar structure to genistein but lacking Tyr kinase inhibiting properties. Serosal but not mucosal epidermal growth factor (EGF) stimulated NaCl absorption. Mucosal genistein but not genistin also altered second-messenger regulation of neutral NaCl absorption, inhibiting the effect of Ca2+ ionophore A-23187 and of serosal EGF but not affecting the transport changes caused by 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP). In contrast, the Cl secretory effects indicated by the increase in short-circuit current for all three agents, A-23187, EGF, and 8-BrcAMP, were inhibited by mucosal genistein. These results suggest that 1) a Tyr kinase is involved in basally stimulating ileal neutral NaCl absorption and brush-border Na(+)-H+ exchange; 2) EGF stimulates NaCl absorption by an effect exerted from the serosal surface, but the effect also involves a brush-border Tyr kinase; 3) brush-border Tyr kinase is involved in the ability of Ca2+ ionophore A-23187 to inhibit neutral NaCl absorption but is not involved in the transport effects of cAMP. This study suggests that Tyr kinase(s) acting over short time periods is involved in stimulation of neutral NaCl absorption and brush-border Na(+)-H+ exchange and also in Ca(2+)-induced inhibition of NaCl absorption. These studies represent the first example of a brush-border Tyr kinase being involved in short-term signal transduction in intestinal epithelial cells.

Integrated actions of renal medullary prostaglandins in the control of water excretion.

The renal medulla is a rich source of prostaglandins. Two cell types, the collecting tubule epithelium and the interstitial cell, have the enzymatic machinery necessary to make prostaglandins. As a result of this localized production and the proximity to highly specialized medullary structures, the prostaglandins can exert potent effects on several aspects of medullary function. At least three physiologic effects have been clearly documented: reduction of vasopressin-dependent osmotic water permeability of the collecting tubule epithelium, enhancement of medullary blood flow, and inhibition of NaCl absorption from the thick ascending limb of Henle's loop. These three functions are the functions that primarily regulate axial solute content and water excretion. The effect of increasing medullary prostaglandin production is to reduce medullary solute content and increase water excretion. Each action in this regard. The medullary prostaglandins thus antagonize the ultimate action of vasopressin. In addition to the direct antagonistic effect on vasopressin-dependent osmotic water flow across the collecting tubule, prostaglandins and vasopressin may have antagonistic effects at other sites. These actions of medullary prostaglandins provide an integrated mechanism for the "fine tuning" of water excretion.

Inhibition of NaCl absorption from perfused rat ileum by furosemide.

The effect of furosemide on intestinal absorption of water and electrolytes was studied using segments of rat ileum perfused in vivo. Furosemide (1 mM) in the perfusion fluid reduced absorption of Na, Cl, and water by 50% from a balanced electrolyte solution without changing the transepithelial potential difference (PD). This effect was also observed in the absence of luminal glucose and was largely reversible. Substitution of all Na in perfusion fluid with choline produced secretion of Na and water and abolished Cl absorption; substitution of all Cl with SO4 reduced Na absorption to 20% of control values. Under both these conditions, furosemide had only trivial effects on electrolyte absorption and exerted no effect on PD. Measurements of unidirectional fluxes of Na and Cl showed that furosemide decreased net flux by reducing lumen-to-blood flux of these ions rather than increasing blood-to lumen flux. These results resemble those obtained in this tissue following exposure to acetazolamide, and suggest that furosemide inhibits a coupled, neutral process of NaCl transport from lumen to blood. Although this effect could be a result of carbonic anhydrase inhibition it more likely occurs from a separate action of furosemide on ileal transport.