PubMed Health⌕ Search

Biomedical subjects

T C Welbourne

Publications and source records attributed to T C Welbourne.

At least 37 records · Page 2Linked to original sources
PubMed1993

Glucocorticoid regulation of splanchnic glutamine, alanine, glutamate, ammonia, and glutathione fluxes.

Interorgan glutamine and associated metabolite fluxes were measured across the gut and liver to delineate splanchnic bed fluxes secondary to enhanced arterial loads mobilized in the periphery by glucocorticoid. Experiments were performed on adrenalectomized rats since adrenalectomy doubled the hepatic glucocorticoid receptor population compared with intact animals. Under these conditions, triamcinolone supplement (40 micrograms.day-1.100 g body wt-1) enhanced the combined net glutamine uptake by gut and liver eightfold, whereas combined gut and liver unidirectional breakdown and synthesis fluxes both increased (3.4- and 7.4-fold, respectively). Triamcinolone supplement also altered the pattern of metabolite released; gut released predominantly ammonium and some alanine, whereas the liver removed more alanine along with glutamine and released more urea, glutamate, and glutathione. Mechanistically, enhanced cellular glutamine uptake could be attributed to a three- to fourfold acceleration of glutamine transport associated with a rise in intracellular glutamine content. However, uptake by isolated membrane vesicles revealed only a small (27%) increase in System N activity, whereas extraction and reconstitution of the transporter into proteoliposomes failed to demonstrate increased transporter activity. Similarly, activity of phosphate-dependent glutaminase and glutamate dehydrogenase increased in crude homogenates (2-fold), but the former disappears in completely disrupted preparations. Furthermore, whereas messenger RNA and assayable enzymic activity for glutamate dehydrogenase clearly increased with glucocorticoid, glutaminase message was less significantly increased. Thus glucocorticoid appears directly capable of accelerating hepatic glutamine extraction primarily by modulating transporter activity that is closely coupled to glutamine utilization.

Adrenalectomy↗
PubMed1993

Growth hormone-enhanced acid production and glutamate and glutamine utilization in LLC-PK-F+ cells.

Growth hormone (GH), recombinant human (rh) GH, effects on transepithelial proton gradient, acid production, and glutamine (Gln) and glutamate (Glu-) utilization were studied in LLC-PK-F+ cells. Monolayers were incubated in Dulbecco's modified Eagle's media containing either 2 mM Gln or Glu- and 5 mM glucose plus 10% fetal bovine serum for 4 h, after which samples were taken for metabolite and acid-base parameter measurements. Monolayers grown on porous supports in Glu- media responded to 32 nM rhGH by decreasing Glu- net uptake 29% (470 +/- 135 to 336 +/- 98 nmol/4 h, P < 0.05), reflecting elimination of basolateral uptake (189 +/- 80 to 2 +/- 35 nmol/4 h, P < 0.05) as ammonium production decreased. In 2 mM Gln, rhGH decreased Gln net uptake 84% (1,096 +/- 215 to 155 +/- 150 nmol/4 h, P < 0.05), with basolateral uptake reversing to release; Glu- formation increased as ammonium production decreased. Monolayers grown on porous supports required rhGH to generate a transepithelial proton gradient within 4 h (0.01 +/- 0.05 control vs. 0.27 +/- 0.11 U for rhGH, P < 0.05); in addition, rhGH increased acid production (16.7 +/- 2.0 to 21.5 +/- 0.6 mumol x 4 h-1 x mg protein-1, P < 0.05). Adding insulin-like growth factor I (IGF-I) to the basolateral media decreased pH and enhanced Glu- and Gln uptake from that compartment.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Glutamine stability in biological tissues evaluated by fluorometric analysis.

Although glutamine has been considered unstable during storage and therefore difficult to quantitate, recent results suggest this amino acid is stable at low pH ranges. We evaluated the stability of glutamine in plasma and tissue extracts, using fluorometric analysis. The measured concentration of glutamine detected varied linearly up to 0.8 mmol/L for the aqueous solution (r2 = 98.7, P = 0.0001) with a mean (+/- SD) coefficient of variation of 2.41% +/- 0.79%. When glutamine was dissolved in 50 g/L trichloroacetic acid (TCA), the values were essentially unaltered. Glutamine in an aqueous solution and stored at -70 degrees C was stable for at least 16 days; glutamine in TCA was stable for 6-8 days, then decreased to a concentration significantly lower than that of the aqueous solution. The expected and observed concentrations in plasma were equal (r2 = 0.99975) for increasing amounts of added glutamine. Glutamine concentrations in plasma were stable for > 1 year when stored at -70 degrees C. The glutamine of a transplantable rat sarcoma and a normal rat liver could be extracted with 50 g/L TCA with high efficiency (88.6% +/- 1.9% and 90.2% +/- 0.04%, respectively); the extracted glutamine is stable in TCA for at least 7 days without neutralization when stored at -70 degrees C. Fluorometric analysis of glutamine required only a small quantity of plasma (25 microL) or tissue (200 mg) and is a convenient method for quantifying this important amino acid.

Drug Stability↗
PubMed1992

Effect of altered acid-base balance and of various agonists on levels of renal glutamate dehydrogenase mRNA.

Rat kidney contains 3.5-kb and 2.8-kb mRNAs that encode for glutamate dehydrogenase (GDH). The levels of both mRNAs are increased gradually after onset of chronic metabolic acidosis and reach a maximum induction of 2.5-fold after 7 days. In contrast, during recovery from chronic acidosis, the levels of the GDH mRNAs are returned to normal within 1 day. The development of an acute metabolic acidosis causes a more rapid induction of GDH mRNA. This increase occurs after a 7-h lag and plateaus after 18 h at a level that is threefold greater than normal. A very similar profile was observed after the transfer of LLC-PK-F+ cells from normal medium to an acidic medium containing 10 mM bicarbonate and adjusted to pH 6.9. However, the transfer of cells from acidic to normal medium caused an immediate and rapid [half-life (t) = 1 h] decrease in GDH mRNA. The apparent half-lives of GDH mRNA were measured by treating cells grown in normal (t = 4 h) and acidic media (t = 12 h) with actinomycin D. Thus, increased stability may account for the induction of GDH mRNA that occurs during growth in response to acidosis. The levels of GDH mRNA are independently affected by changes in medium pH or bicarbonate concentration. The levels of GDH mRNA are also increased by treating cells with adenosine 3',5'-cyclic monophosphate, epinephrine, triiodothyronine, or retinoic acid, whereas treatment with angiotensin II, vasopressin, phorbol 12-myristate 13-acetate, or cycloheximide did not produce an increase. The inductive effect of dexamethasone, which is observed in vivo, is not reproduced in the LLC-PK-F+ cells.

Acid-Base Equilibrium↗
PubMed1992

Growth hormone and renal glutamine and glutamate handling.

Growth hormone administration effects a positive nitrogen balance in part by recycling glutamine nitrogen as glutamate at the expense of ureagenesis. The study presented here focuses on the response of the isolated perfused hypophysectomized rat kidney to acute growth hormone administration during infusions of either glutamine or glutamate. Growth hormone at 50 nM acutely decreases the renal utilization of both glutamine and glutamate while enhancing reabsorption of the latter. During glutamine infusions of either 1,000 or 500 nmol/min, growth hormone markedly reduced net glutamine utilization by 55% at the high loads and reversed utilization to release at the lower load; associated with decreased glutamine utilization was reduced ammonium production and increased glutamate release. Although glutamine reabsorption was unchanged, glutamate reabsorption increased and NH4+ excretion decreased. During glutamate infusion of 180 nmol/min, growth hormone reduced glutamate utilization 66%, the residual utilization matching increased glutamate reabsorption was associated with enhanced bicarbonate reabsorption and a redistribution of NH4+ release into the urine; all three responses were eliminated by amiloride. These responses to growth hormone are consonant with reduced glutamate oxidation underlying decreased glutamine utilization and accelerated luminal Na+-H+ exchange mediating luminal transport, events that are conceivably interrelated.

Amiloride↗
PubMed1991

Growth hormone accelerates tubular acid secretion.

The effect of growth hormone on tubular H+ secretion by the hypophysectomized and intact rat was studied in the isolated functioning kidney. Net acid secretion was estimated as the sum of HCO3- absorption plus NH+4 excretion. Kidneys from either intact or hypophysectomized rats were isolated and perfused over a 90-min time course during which either recombinant human growth hormone (GH), insulin-like growth factor-1 (IGF-1), porcine insulin, or vehicle was added; hormone response was then compared with the time controls. Compared with kidneys from intact rats, hypophysectomized rat kidneys exhibited a marked acidification defect, net H+ secretion, 13,530 +/- 600 vs. 17,860 +/- 810 (SE) nmol/ml of glomerular filtrate (GF). Administering GH (50 nM) increased net H+ secretion within 15 min in both hypophysectomized and intact groups to a maximum of 17,950 +/- 910 and 20,960 +/- 1,100 nmol/ml GF, respectively; neither insulin nor IGF-1 (50 nM) was able to mimic GH's effect. Addition of 1 mM amiloride completely abolished the GH-accelerated acid secretion and greater than 70% of the basal net acid secretion rate. Furthermore, GH-enhanced volume absorption was also abolished by amiloride, although neither NaCl nor glutamine absorption was affected. GH-accelerated acid secretion and coupled volume absorption could be observed at concentrations as low as 3.5 nM with half-maximal effect at 12 nM, which is within the range of GH concentration achieved during episodic GH surges. Finally administering GH in vivo to hypophysectomized rats enhanced net acid secretion and urinary acidification, consistent with accelerated tubular H+ secretion as one physiological expression of GH action.

Amiloride↗
PubMed1990

Interorgan glutamine flow regulation in metabolic acidosis.

The flow of glutamine to the kidneys is essential for generating base in response to acid loading yet neither the magnitude nor direction of this flow are normally supportive of renal ammoniagenesis. However, chronic metabolic acidosis sets in motion regulatory systems enhancing flow magnitude as well as redirecting glutamine from the splanchnic bed and ureagenesis to the kidneys for ammoniagenesis and bicarbonate generation. These mechanisms include organ-specific inductions of glutamine synthesizing and hydrolyzing enzymes at the source, muscle, and the destination, kidneys, respectively; organ-specific shifts in fluxes through competing metabolic pathways favoring glutamine formation at the expense of the ureagenic precursor alanine and unique interorgan regulation whereby upstream sites modulate subsequent downstream sites by setting the glutamine loads and the release of glutamine metabolites acting as metabolic signals. These extrarenal regulatory mechanisms act in concert making glutamine available at the expense of ureagenesis. The kidneys draw upon plasma glutamine, despite a 40% reduction in the arterial concentration, generating base in the form of renal venous bicarbonate and excreting nitrogen and protons as ammonium. Underlying this enormous renal extraction is a shift in the uptake mode from a load- to a transport-limited process closely associated with the filtered bicarbonate load. Finally the interorgan glutamine flow set in motion during acidosis can be acutely reversed, revealing a hierarchal interaction of system subserving acid base and nitrogen balance. Thus, the extraordinary responses exhibited in chronic metabolic acidosis provide a superb model for discerning regulatory systems in other physiological as well as pathophysiological conditions.

Acidosis, Renal Tubular↗
PubMed1989

Atrial natriuretic peptide inhibits renal glutamine extraction.

The effect of administered atrial natriuretic peptide, ANP, on renal glutamine extraction, oxygen consumption and ammoniagenesis was determined in the intact functioning kidney of nonacidotic and chronically acidotic rats. Chronic acidosis shifted the metabolic fuel dependency towards glutamine, reflected by a 3.7 fold increase in extraction. Bolus injection of ANP, 1 microgram/100g BW, results in increased GFR, massive diuresis and an acid urine. Glutamine uptake fell in both groups, reversing from uptake to release in nonacidotic animals and dropping nearly 50 percent in acidotic rats. In contrast, oxygen consumption fell only 20 percent. Inhibition of glutamine extraction appears to be an indirect effect of ANP dependent upon the elevated GFR and elimination of glutamine uptake from the blood, but not from the filtrate. Efficacy of ANP, unlike classical diuretics, was not affected by the prevailing acid base condition nor a large shift in the fuel utilized.

Animals↗
PubMed1989

Glucocorticoid and acid-base homeostasis: effects on glutamine metabolism and transport.

The amino acid glutamine serves as a major fuel, powering tubular transport processes and base generation in the kidneys of acidotic animals. Removing the adrenals limits the kidneys' response to an exogenous acid load and impairs the removal rate of glutamine from the blood. Administering glucocorticoid to adrenalectomized (ADX) rats reverses a small net release of glutamine to uptake; similarly, administering an exogenous acid load to ADX rats restores the kidneys to organs of net extraction. Glucocorticoid-induced uptake is substantially less than the filtered glutamine load in contrast to uptake across both the luminal and basolateral surfaces when filtered HCO3- is reduced. Furthermore, mitochondrial glutamine oxidation is glucocorticoid dependent with a distinct impairment exhibited by the ADX acidotic rat kidney. However, combining the effect of reduced filtered HCO3- with glucocorticoid supplementation results in a marked potentiation of glutamine uptake coupled to oxidation. These results are consistent with a proposed proximal tubule cell model in which basolateral glutamine uptake is inversely related to basolateral HCO3- efflux and coupled, in the presence of glucocorticoid, to mitochondrial oxidation. This model of glucocorticoid-induced ammoniagenesis predicts an increased renal work capacity when basolateral transport of a major metabolic fuel, glutamine, is closely coupled to its oxidative metabolism.

Acid-Base Equilibrium↗
PubMed1989

Hormonal and Na+ effect on renal glutamine uptake.

The flux of glutamine through cellular PDG or extracellular gamma-glutamyltransferase may reflect competition between the membrane glutamine transporter and extracellular hydrolysis. To test this model, kidneys from nonacidotic (NA) and chronically acidotic rats were isolated and perfused with glutamine in a Na+ media and then with a media in which choline replaced Na+. Replacing Na+ abolished the Na+ gradient and completely eliminated the transport of filtered glutamine back into the perfusate. Nevertheless NH4+ production and glutamine utilization remained intact while glutamate accumulated in the extracellular compartments. In the Na+ media, acidotic rat kidneys produced 2.6 times more NH4+ and utilized 1.5-fold more glutamine than their NA counterparts. In the choline media, acidotic rat kidneys had a large drop in NH4+ production but not in glutamine utilization; associated with this was a large glutamate accumulation in the extracellular compartments consistent with flow through the extracellular pathway. To demonstrate the applicability of this model, the role of the adrenals and glucocorticoids in modulating fluxes over these pathways was studied. Kidneys from adrenalectomized acidotic rats exhibited a diminished transport of glutamine, reduced utilization and decreased ammoniagenesis; glutamate accumulation in the extracellular compartment rose consistent with greater flux via the extracellular glutaminase. Supplementing these animals with triamcinolone restored flow into the cellular pathway. The results confirm the in situ activity of two renal glutaminases whose relative contributions to renal ammoniagenesis are dependent on the acid base balance, glucocorticoids, and the existing Na+ gradient.

Acidosis↗
PubMed1988

Gamma glutamyltransferase contribution to renal ammoniagenesis in vivo.

The role of gamma-glutamyltransferase (gamma-GT) in renal ammoniagenesis and glutamine utilization was evaluated in the intact functioning rat kidney. Total NH4+ released, as the sum of renal venous and urinary NH4+, was measured under conditions of chronic metabolic acidosis and paraminohippurate infusion. Ammonia derived from extracellular gamma-GT hydrolysis of glutamine was differentiated from that produced by intracellular phosphate dependent glutaminase (PDG) by employing acivicin, a gamma-GT inhibitor. In non-acidotic animals acivicin administration inhibited gamma-GT 95% and renal venous NH4+ release 48%; NH4+ release into the urine was not inhibited. Chronic metabolic acidosis elevated total NH4+ release 2.5fold, associated with adaptive increase in both gamma-GT and PDG; acivicin reduced total NH4+ released 36% with both renal venous and urinary release effected. The contribution of gamma-GT to total NH4+ production doubles in metabolic acidosis in agreement with the adaptive rise in the in vitro assayed gamma-GT activity. Luminal ammoniagenesis increases in chronic acidosis associated with a fall in urinary glutamine concentration and a rise in the blood to urine glutamine concentration gradient; gamma-GT inhibition eliminates this gradient suggesting luminal ammoniagenesis is largely dependent upon the paracellular glutamine flux. In support of this, paraminohippurate (PAH) infusion increased total renal NH4+ release due entirely to enhanced NH4+ excretion. PAH stimulated luminal ammoniagenesis was associated with an acceleration of renal glutamine extraction and a steeper blood to urine glutamine diffusion gradient; acivicin blocked this response consistent with PAH secretion coupled to activation of intraluminal gamma-GT and glutamine hydrolysis.

Ammonia↗
PubMed1988

Role of glucocorticoids in regulating interorgan glutamine flow during chronic metabolic acidosis.

The role of glucocorticoids in external glutamine mobilization and renal utilization was evaluated in three groups of chronically acidotic rats: sham-treated controls, adrenalectomized, and adrenalectomized supplemented with triamcinolone. Chronic acidosis was induced by administering NH4Cl in their drinking solution over a three-day period. Adrenalectomized rats were supplemented by triamcinolone at a dose of 40 micrograms/100 g/d administered by pellet implantation. Interorgan glutamine flow was evaluated in the postabsorptive state by monitoring net balances across the hindquarters, gut, liver, and kidneys. In the adrenal-intact group, acidosis increased the flow of glutamine from the hindquarters to the kidneys; splanchnic bed uptake, the major glutamine sink in nonacidosis, was eliminated by virtue of hepatic reversal from net uptake to release. Adrenalectomy, in the absence of an exogenous acid load, reversed the flow of glutamine with the kidneys releasing and the hindquarters removing glutamine. Acid loading restored hindquarter glutamine release to levels seen in the intact chronically acidotic animals; however, renal extraction is much less than that exhibited by the intact animals. As a consequence, arterial glutamine concentration rose with the overflow removed by the splanchnic bed, the major glutamine sink in adrenalectomized acidotic rats. Supplementing adrenalectomized acidotic rats with triamcinolone restored glutamine extraction to values seen in intact acidotic rats. Despite the renal extraction, the large hindquarter glutamine release led to hepatic uptake and a high rate of ureagenesis. Glucocorticoids, the release of which is enhanced in metabolic acidosis, appear essential for renal glutamine extraction while playing a lesser role in modulating hindquarter glutamine release.(ABSTRACT TRUNCATED AT 250 WORDS)

Acidosis↗
PubMed1988

Renal ammoniagenic response to chronic acid loading: role of glucocorticoids.

Adrenalectomized (ADX) animals exhibit a blunted renal response to chronic acid loading. To determine whether this response truly reflects impaired renal ammoniagenesis from glutamine, urinary ammonium excretion was compared with acid intake in ADX, intact, and ADX rats supplemented with either a low dose (4 micrograms.100 g-1.day-1) or a high dose (40 micrograms.100 g-1.day-1) of triamcinolone. ADX rats consumed similar amounts of acid as did intact controls yet excreted only 37% of the load as ammonium; in contrast intact controls returned 86% and triamcinolone-supplemented animals returned 98 and 88% for low and high doses, respectively. Nor could the reduced ammonium excretion be attributed to increased renal venous release, since total ammonia production, the sum of renal venous and urine ammonium, was reduced to 49% of the intact controls; low- and high-dose triamcinolone restored and markedly increased the production rate. Underlying the impaired ammonia production rate in ADX rats was a reduced rate of glutamine extraction, 350 +/- 49 vs. 896 +/- 102 and 1,260 +/- 247 and 1,448 +/- 112 nmol.min-1.100 g-1 for intact and low and high doses, respectively. Unlike intact acidotic and glucocorticoid-supplemented ADX acidotic rats, glutamine extraction was disassociated from the delivered glutamine load consonant with the role of glucocorticoid in coupling cellular glutamine transport to its metabolic utilization.

Acid-Base Equilibrium↗
PubMed1987

Renal brush border glutamine transport: comparison between in situ and isolate membrane vesicle uptake.

Glutamine uptake by renal cortical brush-border vesicles was compared to transport expressed by the functioning isolated kidney. Comparisons were made with regard to sodium dependency and the adaptive increase induced by chronic metabolic acidosis in the rat. The results show an absolute dependency upon a sodium gradient; sodium-independent glutamine uptake has no counterpart in situ. In addition, acidosis-induced adaptive increase in vesicle glutamine uptake has no counterpart in situ. Rather, the apparent adaptation reflects extravesicular gamma-glutamyltransferase-mediated conversion to glutamate and subsequent accumulation; acidosis-induced adaptation of this enzyme largely explains the apparent adaptation in glutamine uptake. Consequently the role of membrane transport in glutamine flux regulation can be assessed providing metabolic conversion is controlled.

Acidosis↗
PubMed1987

Gut-liver interaction in glutamine homeostasis: portal ammonia role in uptake and metabolism.

The role of ammonia released by the gut on hepatic glutamine handling and metabolism was studied in postabsorptive anesthesized male Sprague-Dawley rats at spontaneous and elevated arterial glutamine concentrations. Glutamine handling and metabolite release across both organ beds were studied using arteriovenous concentration differences and simultaneously measured portal and hepatic venous plasma flows. At the spontaneous arterial glutamine load, fractional glutamine extraction, FE-Gln, by the gut and the liver was 24 and 10%, respectively. At the elevated glutamine load, gut glutamine uptake doubled, while FE-Gln remained at 24%; however, portal ammonia and alanine increased and decreased, respectively. In response, hepatic FE-Gln increased to 28% with a large release of glutamate and urea. The role of portal ammonia in modulating hepatic glutamine uptake was studied by infusing NH4HCO3 directly into the portal vein. Increasing the portal load promptly stimulated hepatic glutamine uptake and glutamate and urea release. Mitochondria isolated from these livers produced more glutamate from glutamine, suggesting ammonia activation of hepatic glutaminase flux; in addition, citrulline formation increased, suggesting a coupling of glutaminase flux to urea synthesis. The results are consistent with portal ammonia release acting as a key informational molecule in interorgan glutamine flow.

Acid-Base Equilibrium↗