Effects of rat bile infusion on renal function in rats.

The influence of rat bile infusion on renal function in rats and the possible role of bile-induced hemolysis in these effects were examined. The hemolytic action of rat bile and some bile salts were determined in vitro. After the i.v. infusion of rat bile (70 mg freeze-dried powder/2.55 ml) into pentobarbitone-anesthetized rats, the urine, sodium and potassium excretion rates were reduced more than half, which was due to the decrease of glomerular filtration rate and increase of tubular water and sodium reabsorption. A fall in blood pressure, a rise in hematocrit, and hemolysis were also found. Infusion of hemolysed (30 microliters RBC) solution produced by distilled water and then made isotonic caused a short-duration increase in renal excretion and glomerular filtration rate, and the blood pressure was unchanged. Infusion of a rat bile-hemolysed solution after removal of bile acids with cholestyramine increased renal excretions at first with reduction thereafter. Infusion of the rat bile-hemolysed solution treated with barium sulfate produced a renal response very similar to rat bile alone. It is proposed that two factors are involved in the renal response after bile infusion, namely bile acid-induced hemolysis producing diuresis with natriuresis, and bile acid-induced antidiuresis and antinatriuresis, possibly due to a direct renal effect.     

肾盂灌注冲洗对肾脏功能和结构的影响

目的：探讨经皮肾镜碎石术肾盂灌注冲洗压对肾脏结构和功能的影响。方法：建立20头活体猪高压肾盂冲洗模型,建立24F肾造瘘通道,分别在0mmHg（作自身对照,只造瘘不灌注）、150mmHg、200mmHg、250mmHg、300mmHg压力下各冲洗30分钟。术中取肾组织送病理检查,监测肾单位光镜和电镜下的形态学改变;术后5天留取尿标本,应用免疫比浊测定法（ITM）检测尿微量白蛋白（ALB）和β2-微球蛋白（β2-MG）;并于术后第5天再次取肾组织行病理检查观察肾单位的形态学改变。结果：所有灌注组术后都出现尿蛋白的增高,术后第1天和术前相比,都有显著差异（P〈0.01）。形态学观察：当肾盂灌注冲洗压在150-200mmHg时,光镜下观察见肾小囊腔轻度扩张,压力超过250mmHg,肾小囊腔见红细胞和蛋白渗出物,肾小管扩张。电镜下见肾近曲小管上皮细胞内空泡形成,微绒毛排列杂乱、稀疏、部分微绒毛脱落。结论：肾盂灌注冲洗安全压不应超过200mmHg。     

肾盂灌注冲洗对肾脏功能和结构的影响

目的:探讨经皮肾镜碎石术肾盂灌注冲洗压对肾脏结构和功能的影响。方法:建立20头活体猪高压肾盂冲洗模型,建立24F肾造瘘通道,分别在0mmHg(作自身对照,只造瘘不灌注)、150mmHg、200mmHg、250mmHg、300mmHg压力下各冲洗30分钟。术中取肾组织送病理检查,监测肾单位光镜和电镜下的形态学改变;术后5天留取尿标本,应用免疫比浊测定法(ITM)检测尿微量白蛋白(ALB)和β2-微球蛋白(β2-MG);并于术后第5天再次取肾组织行病理检查观察肾单位的形态学改变。结果:所有灌注组术后都出现尿蛋白的增高,术后第1天和术前相比,都有显著差异(P<0.01)。形态学观察:当肾盂灌注冲洗压在150-200mmHg时,光镜下观察见肾小囊腔轻度扩张,压力超过250mmHg,肾小囊腔见红细胞和蛋白渗出物,肾小管扩张。电镜下见肾近曲小管上皮细胞内空泡形成,微绒毛排列杂乱、稀疏、部分微绒毛脱落。结论:肾盂灌注冲洗安全压不应超过200mmHg。     

Effects of thiol antioxidant on reduced nicotinamide adenine dinucleotide phosphate oxidase in hypertensive Dahl salt-sensitive rats

Recent studies implicate of reactive oxygen species (ROS) in hypertension; however, whether reactive oxygen species promote hypertensive derangements is not fully clear. We thus investigated the effects of an antioxidant, N-acetyl-L-cysteine, on hypertensive Dahl salt-sensitive rats. High-salt intake for 4 weeks markedly elevated systolic arterial pressure, urinary excretion of protein, 8-isoprostane, and H(2)O(2), and the enzyme activity of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase along with the elevated expression of its subunits gp91phox and p47phox at the levels of mRNA and protein. Supplement with N-acetyl-L-cysteine reduced the increase in systolic arterial pressure and counteracted the elevation of urinary excretion of protein, 8-isoprostane, and H(2)O(2), and the increases in NADPH oxidase activity/expression in high-salt-loaded Dahl salt-sensitive rats. N-acetyl-L-cysteine supplement ameliorated plasma and urinary levels of thromboxane B(2) (an end metabolite of thromboxane A(2)), associated with improvement of both the abnormal contraction and the impaired nitric oxide-dependent relaxation in renal arteries. These results revealed that oxidative stress mediates hypertensive changes in Dahl salt-sensitive rats, because thiol antioxidant N-acetyl-L-cysteine attenuated the augmentation of local ROS production by diminishing the elevation of NADPH oxidase expression and ameliorated renal/vascular hypertensive changes.     

Relationship between rate of gluconeogenesis and content of nicotinamide adenine dinucleotide in renal cortex

Gluconeogenesis in rat renal cortex was measured using tissue slices incubated with or without appropriate substrates. Immediately after incubation the tissue slices were snap-frozen and the content of oxidized nicotinamide adenine dinucleotide (NAD⁺) was determined. Incubation with 10 mM α-ketoglutarate or L-glutamate led to enhanced glucose production and an increase in tissue content of NAD⁺. Quinolinate and 3-mercaptopicolinate inhibited the rate of gluconeogenesis from L-glutamate and α-ketoglutarate respectively, and decreased the tissue levels of NAD⁺. The enhanced rate of gluconeogenesis was associated with an increase of NAD⁺ in the cytosol fraction (10⁵ × g supernatant) but not in the particulate fraction (10⁵ × g pellet) of renal cortex homogenate. Present results indicate that NAD⁺ content changes in parallel with the rate of gluconeogenesis in renal cortical tissue.     

Light-induced conversion of nicotinamide adenine dinucleotide to nicotinamide adenine dinucleotide phosphate in higher plant leaves

Light-induced conversion of NAD to NADP was investigated in higher plants. Upon illumination, conversion of NAD to NADP was observed in intact leaves of wheat and pea following incubation in the dark. This conversion was also observed in mesophyll protoplasts of wheat leaves when they were isolated in the dark or isolated in light and then preincubated in the dark. Chloroplasts isolated from wheat protoplasts prepared in the dark carried out the conversion. The conversion in the mechanically isolated spinach chloroplasts was observed only when they were isolated in the dark from leaves preincubated in darkness.     

Influence of renal denervation on chronopharmacology of furosemide in rats.

Our previous studies have suggested that the adrenergic nervous system is involved in the mechanism responsible for the time-dependent change in the urinary excretion of furosemide in rats. To examine a potential role of renal nerves in this phenomenon, renal denervation or sham operation was performed using unilaterally nephrectomized rats. Furosemide (30 mg/kg) was given orally at 12 am or 12 pm. Urine was collected for 8 hours after furosemide dosing, and urinary excretions of furosemide and sodium were determined. Urinary furosemide excretion and diuretic effects of the agent (urine volume and urinary sodium) were significantly greater at 12 am than at 12 pm in the sham-operated group of rats. However these administration time-dependent changes in urinary furosemide and its diuretic effects disappeared in the renal-denervated group of animals. These results suggest that the renal nerves contribute to the time-dependent changes in the urinary excretion of furosemide and its subsequent diuretic effects.     

Effect of nicotinamide adenine dinucleotide on the membrane-associated reduced nicotinamide adenine dinucleotide oxidase of Mycobacterium phlei.

NAD+ had a biphasic effect on the NADH oxidase activity in electron transport particles from Mycobacterium phlei. The oxidase was inhibited competitively by NAD+ at concentrations above 0.05 mM. NAD+ in concentrations from 0.02 to 0.05 mM resulted in maximum stimulation of both NADH oxidation and oxygen uptake with concentrations of substrate both above and below the apparent K-M. Oxygen uptake and cyanide sensitivity indicated that the NAD+ stimulatory effect was linked to the terminal respiratory chain. The stimulatory effect was specific for NAD+. NAD+ was also specific in protecting the oxidase during heating at 50 degrees and against inactivation during storage at 0 degrees. NAD+ glycohydrolase did not affect stimulation nor heat protection of the NADH oxidase activity if the particles were previously preincubated with NAD+. Binding studies revealed that the particles bound approximately 3.6 pmol of [14C1NAD+ per mg of electron transport particle protein. Although bound NAD+ represented only a small fraction of the total added NAD+ necessary for maximal stimulation, removal of the apparently unbound NAD+ by Sephadex chromatography revealed that particles retained the stimulated state for at least 48 hours. Further addition of NAD+ to stimulated washed particles resulted in competitive inhibition of oxidase activity. Desensitization of the oxidase to the stimulatory effect of NAD+ was achieved by heating the particles at 50 degrees for 2 min without appreciable loss of enzymatic activity. Kinetic studies indicated that addition of NADH to electron transport particles prior to preincubation with NAD+ inhibited stimulation. In addition, NADH inhibited binding of [14C]NAD+. The utilization of artificial electron acceptors, which act as a shunt of the respiratory chain at or near the flavoprotein component, indicated that NAD+ acts as at the level of the NADH dehydrogenase at a site other than the catalytic one resulting in a conformational change which causes restoration as well as protection of oxidase activity.     

Carbanicotinamide adenine dinucleotide: synthesis and enzymological properties of a carbocyclic analogue of oxidized nicotinamide adenine dinucleotide

The dinucleotide carbanicotinamide adenine dinucleotide (carba-NAD), in which a 2,3-dihydroxycyclopentane ring replaces the beta-D-ribonucleotide ring of the nicotinamide ribonucleoside moiety of NAD, has been synthesized and characterized enzymologically. The synthesis begins with the known 1-aminoribose analogue (+/-)-4 beta-amino-2 alpha,3 alpha-dihydroxy-1 beta-cyclopentanemethanol. The pyridinium ring is first introduced and the resultant nucleoside analogue specifically 5'-phosphorylated. Coupling the racemic carbanicotinamide 5'-mononucleotide with adenosine 5'-monophosphate produces two diastereomeric carba-NAD analogues which are chromatographically separable. Only one diastereomer is a substrate for alcohol dehydrogenase and on this basis is assigned a configuration analogous to D-ribose. The reduced dinucleotide carba-NADH was characterized by fluorescence spectroscopy and found to adopt a "stacked" conformation similar to that of NADH. The analogue is reduced by both yeast and horse liver alcohol dehydrogenase with Km and Vmax values for the analogue close to those observed for NAD. Carba-NAD is resistant to cleavage by NAD glycohydrolase, and the analogue has been demonstrated to noncovalently inhibit the soluble NAD glycohydrolase from Bungarus fasciatus venom at low concentrations (less than or equal to 100 microM).     

Hormonal interactions and renal function during mechanical ventilation and ANF infusion in humans

To investigate the influence of atrial natriuretic factor (ANF) on renal function during mechanical ventilation (MV), we examined the renal and hormonal responses to synthetic human ANF infusion in eight patients during MV with zero (ZEEP) or 10 cmH2O positive end-expiratory pressure (PEEP). Compared with ZEEP, MV with PEEP was associated with a reduction in diuresis (V) from 208 +/- 51 to 68 +/- 11 ml/h (P less than 0.02), in natriuresis (UNa) from 12.4 +/- 3.3 to 6.2 +/- 2.1 mmol/h (P less than 0.02), and in fractional excretion of sodium (FENa) from 1.07 +/- 0.02), 0.21 to 0.67 +/- 0.17% (P less than 0.02) and with an increase in plasma renin activity (PRA) from 4.83 +/- 1.53 to 7.85 +/- 3.02 ng.ml-1.h-1 (P less than 0.05). Plasma ANF levels markedly decreased during PEEP in four patients but showed only minor changes in the other four patients, and mean plasma ANF levels did not change (163 +/- 33 pg/ml during ZEEP and 126 +/- 30 pg/ml during PEEP). Glomerular filtration rate and renal plasma flow were unchanged. Infusion of ANF (5 ng.kg-1.min-1) during PEEP markedly increased V and UNa by 110 +/- 61 and 107 +/- 26%, respectively, whereas PRA decreased from 7.85 +/- 3.02 to 4.40 +/- 1.5 ng.ml-1.min-1 (P less than 0.05). In response to a 10 ng.kg-1.min-1 ANF infusion, V increased to 338 +/- 79 ml/h during ZEEP but only to 134 +/- 45 ml/h during PEEP (P less than 0.02), whereas UNa increased, respectively, to 23.8 +/- 5.3 and 11.3 +/- 3.3 mmol/h (P less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)     

Protein binding of nicotinamide adenine dinucleotide and regulation of nicotinamide adenine dinucleotide glycohydrolase activity in homogenates of rabbit skeletal muscle

Gel-permeation chromatography and ultrafiltration have been used to study the free and bound forms of NAD in crude extracts prepared from rabbit muscle. Both techniques indicate that over 80% of the endogenous NAD is free.Nicotinamide inhibits the destruction of NAD in muscle homogenates (50% inhibition at 1.6 mm nicotinamide). In the absence of nicotinamide, there is a rapid destruction of free NAD, but a more gradual destruction of bound NAD. The latter result confirms earlier findings that bound NAD is protected from the hydrolytic action of NADase. However, this protection is unlikely to constitute an important mechanism for controlling NADase activity in muscle homogenates because such a small proportion of the endogenous NAD is bound.In the absence of nicotinamide, NAD also disappears rapidly from minced muscle. Interestingly, the NAD/NADH ratio remains constant (NAD/NADH = 18.1–18.5) during the disappearance of NAD in minced muscle. Upon homogenization of the mince, the NAD/NADH ratio abruptly decreases, then slowly increases during subsequent incubation. The latter rise in NAD/NADH ratio appears to be independent of absolute changes in NAD concentration brought about by the action of NADase or the addition of exogenous NAD.     

Effects of dietary manipulations and glucose infusion on glucagon response during exercise in rats

Tadjoré, Maurice, Raynald Bergeron, Martin Latour,François Désy, Claude Warren, and Jean-Marc Lavoie.Effects of dietary manipulations and glucose infusion on glucagonresponse during exercise in rats. J. Appl.Physiol. 83(1): 148-152, 1997.The purpose of thepresent investigation was to test the hypothesis that blood glucoseconcentration is not always related to glucagon response duringexercise. Three groups of rats were submitted to a prolonged (3-h)swimming exercise. Two groups of rats had their normal food intakerestricted by 50% the night before the experiment. One of these twogroups of rats was intravenously infused with glucose throughoutexercise to maintain euglycemia. The third group of rats swam whileunder normal dietary conditions. Plasma glucose, sampled in arterialblood, was reduced (P < 0.05) at 75, 105, 150, and 170 min of exercise (from ~130 to 110 mg/dl) in thefood-restricted animals without glucose infusion, whereas a significant(P < 0.05) increase was measured inthe two other groups during exercise. A significant(P < 0.01) difference in the meanintegrated areas under the glucose-concentration curve was found onlybetween the fed and the two food-restricted groups. Plasma insulinconcentrations decreased (P < 0.05)similarly in all groups during exercise, whereas plasma epinephrine andnorepinephrine concentrations increased significantly(P < 0.01) in all groups. Despitedifferences between groups in plasma glucose response during exercise,and despite the absence of any decrease in exercising blood glucoselevels in at least two of the three groups, plasma glucagon responseswere increased (P < 0.05) similarlyin all groups (from ~250 to 550 pg/ml) at the end of the exerciseperiod. The increase in glucagon was significant after 90 min ofexercise in the food-restricted groups, with or without glucoseinfusion, but only after 140 min in the fed group. These resultsindicate that the glucagon response during exercise is not alwayslinked to the decrease in plasma glucose.

     

Coupling between reduced nicotinamide adenine dinucleotide oxidation and metabolite transport in renal brush border membrane vesicles

In a previous communication [Giménez-Gallego, G., Benavides, J., García, M.L., & Valdivieso, F. (1980) Biochemistry (preceding paper in this issue)] we have reported the occurrence of a NADH oxidase activity in the renal brush border membranes. The brush border membranes can utilize the energy from the oxidation of NADH to drive the transport of amino acids (aspartic and glutamic acids), organic acids, and the lipophilic cation tetraphenylphosphonium (TPP). The coupling between NADH oxidation seems to be due to the formation of a proton electrochemical gradient (delta-mu H+) as indicated by the effect of specific ionophores. This system may be implicated in the reabsorption process in the renal tubules and in the maintenance of the delta-mu H+ (positive and acidic in the luminal side) previously described in the renal tubules "in vivo".