The effect of suppression of prostaglandin synthesis on renal function in rats with intact and reduced renal mass

The present study was undertaken to assess the role of prostaglandin system in the compensatory response to reduced nephron population, respective to renal function and electrolyte excretion. Intact and nephrectomized rats were divided in 4 groups: 1) rats pretreated with indomethacin, 2) rats pretreated with the vehicle of indomethacin, 3) rats pretreated with sulindac, and 4) rats pretreated with the vehicle of sulindac.In normal rats, indomethacin administration resulted in a mild decrease in creatinine clearance and a significant reduction of the urinary Na excretion. In the rats with reduced renal mass treated with indomethacin, the creatinine clearance did not differ from that in the control group. The 24 h urinary sodium excretion and the fractional excretion of sodium, however, were significantly lower in the indomethacin treated animals than in the control rats. No change in the creatinine clearance or in the sodium excretion was observed in all groups pretreated with sulindac.The urinary PGE₂ and thromboxane excretion was significantly lower in the indomethacin treated intact rats and the rats with reduced renal mass. Sulindac induced a slight decrease in urinary excretion of PGE₂ in intact rats. No significant change in urinary excretion of PGE₂ or thromboxane was seen after sulindac in the rats with reduced renal mass.The antinatriuretic effect of indomethacin was dissociated from changes in urine flow in all groups of animals, suggesting that the increase in Na reabsorption tool place in a water impermeable segment of nephron.These results suggest that the compensatory increase in urinary Na excretion per nephron in rats with reduced nephron population at least partly depends on an intact prostaglandin synthesis.     

Effect of glutathione manipulation on prostaglandin synthesis in renal medullary homogenates.

1. The effect of glutathione (GSH) manipulation on arachidonic acid (AA) metabolism in renal medullary (RM) homogenates was investigated. 2. Diethyl maleate (DEM) depleted GSH initially by 50% (P less than 0.05) and produced a general suppression (P less than 0.05) of all PGs with the exception of TXB2. GSH was further depleted during homogenization and a 30-min incubation period (P less than 0.01). 3. Adding glutathione monoethyl ester (GSH-MEE) (0, 0.8, 1.6 or 3.2 mmol/ml) to RM homogenates increased GSH (P less than 0.01) and decreased RM homogenates' PGs-synthesizing capability (P less than 0.05), with the exception of PGE2 and TXB2 at the highest concentration. 4. The results indicate that homogenization has a significant impact (P less than 0.05) on GSH concentration of the media and alterations in GSH concentration affect the profile and quantity of AA metabolites in renal medullary homogenates.     

Influence of dietary polyunsaturated fatty acids on renal & aortic prostaglandin synthesis in 1 kidney 1 clip goldblatt hypertensive rats

To study the influence of dietary modification on prostaglandin synthesis and on blood pressure regulation, the effects of dietary enrichment with linolenic or linoleic acid was compared with standard rat chow in 3 groups of 13 rats before and after renal artery constriction and contralateral nephrectomy. Before renal artery constriction 4 weeks supplementation with 40 en% linseed oil (53% linolenic acid) increased renal linolenic acid, decreased arachidonic acid, and suppressed synthesis of 6-keto-PGF_1α and PGE₂ by renal homogenates (33% and 38% respectively, p<0.01) compared with standard diet. Rats fed on 40 en % sunflower seed oil (63% linoleic acid) increased renal prostaglandin synthesis (p<0.05) compared with linseed oil, but not compared with standard diet. Seven weeks after renal artery constriction renal and aortic 6-keto-PGF_1α and PGE₂ were suppressed 30% to 50% (p<0.05) by linseed oil supplements compared with sunflower seed oil and standard diets. In the sunflower seed oil group aortic 6-keto-PGF_1α correlated (r = 0.75, p<0.02) with final systolic blood pressure. Final systolic blood pressures were similar in linseed oil (152.9 mmHg ± se 3.3, sunflower oil (155.1 ± se 6.6) and standard diet group (159.0 ±se 4.2). Thus dietary linseed oil suppressed renal and aortic prostaglandin synthesis but did not accentuate renal hypertension, and linoleic acid supplementation did not protect against 1 kidney 1 clip renal hypertension.     

Effects of vasopressin and prostaglandin synthesis inhibition on cyclic AMP accumulation in the dog renal inner medulla

The interaction of arginine vasopressin (AVP) and endogenous prostaglandins on cAMP production was investigated in the dog. Cyclic AMP content of dog inner medullary tissue slices exposed to different concentrations of AVP in the presence and absence of various prostaglandin synthesis inhibitors was determined. If the slices were incubated in isotonic media with 95% O2 and 5% CO2 gas phase, inhibition of prostaglandin synthesis decreased cAMP accumulation. A significant correlation was found between the decrements in cAMP content and basal cAMP levels. AVP-induced increments in cAMP accumulation was, however, unaffected by prostaglandin synthesis inhibitors. If incubation was performed in a hypertonic medium and at low O2 concentration, basal cAMP content was significantly reduced and it was not altered by inhibition of prostaglandin synthesis. The cAMP response to AVP was practically identical in the presence and absence of prostaglandin synthesis inhibitors. In conscious dogs AVP and indomethacin in itself had no effect on urinary cAMP excretion, but there was a significant decrease if the two compounds were combined. These results fail to lend support the hypothesis that endogenous prostaglandins modulate AVP-induced cAMP accumulation in the inner medulla.     

The effect of inhibition of prostaglandin synthesis on ovarian hypertrophy in the rat.

Aspirin and indomethacin, inhibitors of prostaglandin biosynthesis, were utilized to determine the role of prostaglandins (PGs) in ovarian weight gain in rats following unilateral ovariectomy or treatment with PMSG. After unilateral ovariectomy, the compensatory ovarian hypertrophy was 185-0% compared with 139-8% and 97-5% in rats treated with indomethacin and aspirin, respectively. The adrenal weights in rats treated with aspirin were also reduced significantly. Administration of PGE2 or PGF2alpha with aspirin reversed the effect of aspirin on the adrenals but had no effect on the ovarian weight. Indomethacin and aspirin treatment of animals injected with PMSG also reduced the ovarian weight gain. If 100 mug PGE2 were given twice daily, this effect was reversed in both groups but thrice daily administration had no effect on rats receiving aspirin. In PMSG-treated rats, 100 mug PGF2alpha twice daily did not reverse the effect of indomethacin and aspirin, and actually enhanced the effect of aspirin.     

Age and renal prostaglandin inhibition during exercise and heat stress.

Aging is associated with a number of physiological changes that may cause the kidney to rely to a greater extent on vasodilatory PGs for normal functioning. Acute exercise has been shown to cause renal vasoconstriction that may be partially buffered by vasodilatory PGs. To determine the relative importance of renal PGs during exercise in older adults, we compared the renal effects of the PG inhibitor ibuprofen (1.2 g/day for 3 days) vs. a placebo control in a cohort of eight younger (24 +/- 2 yr) and eight older (64 +/- 2 yr) women during treadmill exercise ( approximately 57% maximal oxygen consumption) in the heat (36 degrees C). This over-the-counter dose of ibuprofen reduced renal PG (i.e., PGE2) excretion by 47% (P < 0. 05). Acute exercise in the heat caused dramatic decreases in glomerular filtration rate, renal blood flow, and sodium excretion in both age groups. PG inhibition was associated with greater decreases in urine production and free water clearance (P < 0.05). There were no drug-related declines in glomerular filtration rate or renal blood flow. We conclude that PG inhibition has only modest effects on renal function during exercise. Also, the lack of hemodynamic changes with PG inhibition indicates that healthy well-hydrated older women are not in a renal PG-dependent state.     

Interferon-mediated inhibition of prostaglandin synthesis in human mononuclear leukocytes

In this study, the question of whether human leukocyte-derived and fibroblast-derived interferon had an effect on prostaglandin metabolism in human peripheral blood mononuclear cells has been considered. Both leukocyte- and fibroblast-derived interferon were potent inhibitors of mononuclear cell prostaglandin synthesis at low physiological concentrations. Inhibition required a minimum incubation of 1 hr. Interferon had no effect on release of arachidonic acid; synthesis of hydroxy fatty acids was slightly increased.     

Influence of renal denervation on renal effects of acute nitric oxide and ETA/ETB receptor inhibition in conscious normotensive rats.

The role of renal nerves in the effects of concomitant NO synthase and non-selective ET(A/)ET(B) receptor inhibition on renal function was investigated in conscious normotensive Wistar rats. NO synthase inhibition alone (10 mg/kg b. w. i.v. L-NAME) in sham-operated rats with intact renal nerves induced an increase in systolic, diastolic and mean arterial pressure, urine flow rate, sodium, chloride and calcium excretion (p<0.05). The effect of L-NAME was markedly reduced by bosentan (10 mg/kg b.w. i.v.) and the values of urine flow rate, sodium, chloride and calcium excretions returned to control level (p<0.05). L-NAME administration one week after a bilateral renal denervation increased blood pressure to a similar extent as in sham-operated rats but decreased urine flow rate (p<0.05) and did not change electrolyte excretion. ET(A/)ET(B) receptor inhibition with bosentan during NO synthase inhibition in the renal denervated rats did not produce changes in urine flow rate or electrolyte excretion. NO synthase inhibition as well as concurrent NO synthase and ET(A/)ET(B) receptor inhibition did not change clearance of inulin or paraaminohippuric acid in sham-operated or renal denervated rats. These results indicate that renal sympathetic nerves play an important modulatory role in NO and endothelin induced effects on renal excretory function.     

Effect of prostaglandin inhibition on renal response to volume expansion in dog

A prostaglandin (Pg) has been proposed as a “natriuretic hormone” effecting the natriuresis observed with extracellular fluid volume expansion. Dogs were volume expanded under chloralose anesthesia by equilibrating the animal's blood with the saline expanding volume outside of the body prior to infusion, in order to avoid hemodilution during expansion. The massive natriuresis associated with expansion was not significantly affected by preloading injection of indomethacin (5–10 mg/kg) to depress prostaglandin synthesis. Consistent with studies by others, a rise in filtration fraction was observed in 4 of 5 experiments with indomethacin but not in its absence. The results do not support the concept that the hypothetical natriuretic hormone is a Pg or is dependent upon a Pg for its synthesis or release. The data, moreover, constitute further evidence that the nutriuresis of acute volume expansion does not depend upon decreased colloid osmotic pressure in the renal peritubular capillaries. Urinary Pg F_2αexcretion was depressed by indomethacin but was unaffected by expansion. Expansion was associated with an unexplained 10 percent decrease in plasma potassium concentration.     

Effect of prostaglandin synthesis inhibition on human corpus luteum function

The hypothesis that human luteolysis is mediated by prostaglandin F_2α was initially tested through placebo-controlled observations of the effect of a single dose of ibuprofen on progesterone and luteinizing hormone levels late in the luteal phase in normal human subjects. Falling levels of progesterone at rest, not significantly attenuated by ibuprofen, did not support, and falling progesterone/luteinizing hormone ratios at rest, significantly attenuated by ibuprofen, did support this hypothesis. The pulsatile nature of progesterone levels in the human was confirmed in this study.     

Effect of prostaglandin synthesis inhibition on human corpus luteum function

The hypothesis that human luteolysis is mediated by prostaglandin F2 alpha was initially tested through placebo-controlled observations of the effect of a single dose of ibuprofen on progesterone and luteinizing hormone levels late in the luteal phase in normal human subjects. Falling levels of progesterone at rest, not significantly attenuated by ibuprofen, did not support, and falling progesterone/luteinizing hormone ratios at rest, significantly attenuated by ibuprofen, did support this hypothesis. The pulsatile nature of progesterone levels in the human was confirmed in this study.     

The effect of prostaglandin synthesis inhibition on the direct stimulation of renin release from rabbit renal cortical slices

Prostaglandins have been hypothesized to have several mechanistic functions in sympathetically mediated release of renin. The rabbit renal cortical slice system was chosen to examine the prostaglandin dependency of renin release directly stimulated by either a direct adenylate cyclase activator, forskolin, or a β-agonist, isoproterenol. In this study, we demonstrate that with forskolin (1 × 10⁻⁵M) or isoproterenol (1 × 10⁻⁶M), renin release was elevated 2–3 fold above control, and that this increase was shown to accompany a substantial increase in the tissue levels f cAMP (19.5 fold and 3.5 fold respectively). We also demonstrate that the increase in renin release produced by these compounds was not inhibited by cyclooxygenase inhibitors, indomethacin (25 uM) or eicosatetraynoic acid (30 ug/ml), nor was it inhibited by the selective prostacyclin synthesis inhibitor, U-51605 (30 ug/ml). Each of these inhibitors was demonstrated to block the synthesis of prostaglandins in the cortical slices at the concentrations used. Thus we propose that prostaglandins do not play a role in the induction of renin release resulting from elevated cyclic nucleotide levels or β-adrenergic stimulation.     

WR-2721 inhibition of radiation-induced prostaglandin excretion in rats

Pre-irradiation administration of the radioprotectant drug WR-2721 to rats resulted in a significant reduction in radiation-induced increases in excretion rates of prostaglandins (PGE and PGF2 alpha) and thromboxane (TxB2). In animals not irradiated. WR-2721 did not significantly alter these excretion rates. Dramatic reductions in the levels of urinary PGE and TxB2 were observed following exposure to 9.0 Gy of whole-body, unilateral gamma-radiation in WR-2721-treated animals, whereas changes in PGF2 alpha levels were less pronounced. Radiation-induced diuresis was also significantly depressed in animals given WR-2721 before irradiation. Reduced prostaglandin excretion rates may reflect the general radioprotective capacity of the chemoprotector WR-2721 on the release of prostaglandins from radiation-damaged tissue. The decrease in diuresis may be related to the observed prostaglandin decreases.     

The effect of prostaglandin synthesis inhibition on the direct stimulation of renin release from rabbit renal cortical slices

Prostaglandins have been hypothesized to have several mechanistic functions in sympathetically mediated release of renin. The rabbit renal cortical slice system was chosen to examine the prostaglandin dependency of renin release directly stimulated by either a direct adenylate cyclase activator, forskolin, or a beta-agonist, isoproterenol. In this study, we demonstrate that with forskolin (1 X 10(-5) M) or isoproterenol (1 X 10(-6) M), renin release was elevated 2-3 fold above control, and that this increase was shown to accompany a substantial increase in the tissue levels of cAMP (19.5 fold and 3.5 fold respectively). We also demonstrate that the increase in renin release produced by these compounds was not inhibited by cyclooxygenase inhibitors, indomethacin (25 microM) or eicosatetraynoic acid (30 micrograms/ml), nor was it inhibited by the selective prostacyclin synthesis inhibitor, U-51605 (30 micrograms/ml). Each of these inhibitors was demonstrated to block the synthesis of prostaglandins in the cortical slices at the concentrations used. Thus we propose that prostaglandins do not play a role in the induction of renin release resulting from elevated cyclic nucleotide levels or beta-adrenergic stimulation.     

Expression of renal transport systems for inorganic phosphate and sulfate in Xenopus laevis oocytes.

As a first step within an experimental strategy (expression cloning) leading to the structural identification of the two brush-border membrane transport systems for phosphate and sulfate, we have studied the expression of Na(+)-dependent uptake of phosphate and sulfate in Xenopus laevis oocytes injected with rabbit kidney cortex poly(A)+ RNA (mRNA). Na(+)-dependent uptake of phosphate and sulfate was stimulated in a dose- and time-dependent manner up to 20-fold as compared to water-injected controls. After fractionation of the mRNA on a sucrose gradient (or by preparative gel electrophoresis), two neighboring fractions were identified to stimulate Na(+)-dependent phosphate uptake (average size: 3.4 kilobases) and Na(+)-dependent sulfate uptake (average size: 3.7 kilobases). The two transport systems can be discriminated by their inhibition by thiosulfate, which reduced sulfate uptake, but not phosphate uptake. Kinetic characterization of the expressed Na(+)-dependent transport activities results in properties similar to those described for transport activity in renal brush-border membrane vesicles.     

Histamine tachyphylaxis in canine airways despite prostaglandin synthesis inhibition

Tachyphylaxis to aerosolized histamine was studied in dogs anesthetized with thiamylal after pretreatment with prostaglandin synthesis inhibitors. Three consecutive histamine dose-response curves were obtained in nine dogs pretreated with 5 mg/kg indomethacin; two of these nine were also pretreated with 10 mg/kg indomethacin. Seven of the nine dogs were pretreated with 4 mg/kg sodium meclofenamate; four of these seven were also pretreated with 12 mg/kg. All dogs had tachyphylaxis at high concentrations of histamine regardless of inhibitor used. Pretreatment with indomethacin while the dogs were under alpha-chloralose-urethan anesthesia gave similar results. Histamine tachyphylaxis was also studied both in the presence and in the absence of indomethacin in tracheal smooth muscle strips obtained from seven additional dogs. A decrease in the median effective dose to histamine was observed in the indomethacin-treated strips, but tachyphylaxis to histamine remained. We conclude that prostaglandin synthesis inhibition does not reverse histamine tachyphylaxis either in vivo or in vitro. Thus the mechanism of histamine tachyphylaxis remains unexplained.     

Effects of inhibition of prostaglandin synthesis on uterine oxytocin receptor concentration and myometrial gap junction density in parturient rats

The development of oxytocin (OT) sensitivity in the parturient uterus is associated with increases in myometrial OT receptor concentration, gap junction formation, and prostaglandin (PG) production. To investigate whether PGs mediate these responses, we measured OT responsiveness, OT receptor concentrations, and gap junction formations in uteri of Day 19, 20, 21, 22, 23 pregnant and Day 2 postpartum rats. Inhibition of endogenous PG synthesis was produced by infusion of naproxen sodium delivered by an implanted osmotic pump. Naproxen treatment, but not placebo treatment, markedly attenuated in vitro uterine PGE2, PGF2 alpha, and PGI2 releases, suppressed OT responsiveness, and prolonged gestation. The increase of OT receptor concentration that normally occurred on Day 23 term pregnancy was delayed to Day 24. Co-administration of PGF2 alpha reversed the suppressive effects of naproxen. Naproxen treatment did not significantly affect gap junction formations on Day 23 but appeared to delay both the onset and disappearance of gap junction formations. PGF2 alpha co-administration with naproxen also had no apparent effect on gap junction development. The inhibition of OT receptor formation but not gap junction formation on Day 23 in the presence of naproxen indicates that these two events are controlled independently. Furthermore, the failure of naproxen-treated rats to deliver at term suggests that gap junction formation in the absence of an increase in OT receptors is insufficient to initiate labor. It appears that increases in both OT receptor concentrations and gap junction densities may be required for labor.     

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.