Hydropenic Rats Research Articles

Hydropenia may accelerates the progression of orthotopic bladder cancer induced by N-methyl-N-nitrosourea by increasing the expression levels of AQP1, AQP3, and AQP4.

BackgroundIncreasing evidence has demonstrated aquaporins (AQPs) to be critical players in carcinogenesis. Here, we aimed to explore the role of hydropenia in the progression of bladder cancer (BCa), as well as to assess the expression of AQP1, AQP3, and AQP4 in bladder tissues from hydropenic and N-methyl-N-nitrosourea (MNU)-treated rats.MethodsAn orthotopic BCa model was induced by administering Sprague Dawley rats with MNU. A hydropenic rat model was established by administrating rats with 2/3 of the amount of water given to the control group. At week 8, the rats were sacrificed and their bladder tissues were collected. Then, pathological alterations in the rat bladders were assessed by hematoxylin and eosin staining. The RNA and protein expression levels of AQP1, AQP3, and AQP4 were determined by using qRT-PCR and western blot assays.ResultsAll of the rats (100%) administrated with MNU developed tumors, of which 5 were large (diameter, 0.5–1.0 cm), 10 were medium (diameter, 0.2–0.5 cm), and 5 were small (diameter, <0.2 cm) in size. The tumors were nodular and cauliflower shaped, with multiple satellite focus, and were accompanied by bleeding, ulcers, stones, and residual urine. Hematoxylin and eosin staining revealed that the bladder mucosa was incomplete, with a large amount of necrotic tissue and obvious leukocytic infiltration. The tumor volume in the MNU + hydropenia group was significantly larger than that in the MNU group. Noticeably, hydropenia exacerbated pathological changes induced by MNU administration. QRT-PCR and western blot analysis revealed that the MNU group, hydropenia group, and MNU + hydropenia group had significantly increased levels of AQP1, AQP3, and AQP4 compared to the control group, with the most dramatic increase seen in the MNU + hydropenia group.ConclusionsHydropenia exacerbates pathological alterations induced by MNU in rats with orthotopic BCa by increasing the expression levels of AQP1, AQP3, and AQP4. This study reveals a possible mechanism of the occurrence of BCa.

Renal Interstitial Injection of a‐Trinositol Increases Renal Interstitial Hydrostatic Pressure (RIHP) but Does Not Modify its Low Frequency Oscillatory Pattern

RIHP increases after renal interstitial medullary injection (RIMI) of a 5μ bolus of saline solution (SS) + 2 μM α‐Trinositol (α –T, drug that increases collagen fiber network contraction, FASEB J 31:701.4: 2017). To assess whether this increase is induced by changes in the frequency and/or amplitude of oscillatory pattern of RIHP we evaluate in hydropenic rats the time course of RIHP before and after either an acute sham (CTR group, n = 9) or an acute (1s) real RIMI of a 5μ bolus of either vehicle (0.9% SS, SS group, n = 9) or SS + 2 μM α ‐ T (α – T group, n = 9). Left subcapsular RIHP (RIHP) was continuously measured (1 Hz) in anesthetized rats with right nephrectomy, acute left renal denervation, hormonal clamp and renal perfusion pressure control (100 mmHg), before (baseline phase = BL = 30 min) and 30 min after (experimental phase = EXP) sham or real RIMI of 5 μl bolus of either SS or SS + α‐T. The table shows the 30 min average BL RIHP (1800 s, s = samples or seconds), ΔRIHP (EXP RIHP – BL RIHP) values (mean ± SEM) obtained 30 min after sham or real RIMI during 300 s. It also shows the RIHP frequency (F) of major oscillations (cycles/min = cpm) and the standard deviation (SD, an oscillatory amplitude index) of global RIHP oscillation before and 30 min after RIMI. Three were no differences among groups in BL RIHP, F or SD. For ΔRIHP data there were time * group interaction effect (P &lt; 0.009); * P = 0.05, ** P &lt; 0.001 vs. BL; • P &lt; 0.04 vs. SS group, ¨P &lt; 0.001 vs. CTR group. For SD data ° P &lt;0.001 between phases.CONCLUSIONSThe results show that RIHP oscillates at an equally low frequency (≈ 0.03 Hz) before and after sham or real RIMI but with lower amplitude after sham or real RIMI in any of the groups. This suggests that α‐T increases RIHP but not through modify the frequency or amplitude of its low frequency oscillatory pattern.Support or Funding InformationOwn financial resources of Integrative Physiology Lab. RIHP RIHP RIHP (mmHg) Frequency SD Delta cpm (mmHg) BL EXP‐BL BL EXP BL EXP 30 min at min 30 last 5 min at min 30 30 min at min 30 Group 1800 s 300 s 300 s 300 s 1800 s 300 s CTR 3.0 ± 0.10 −1.1 ± 0.16 2.2 ± 0.26** 2.1 ± 0.26 0.54 ± 0.07 0.20 ± 0.04° SS 3.1 ± 0.03 −0.15 ± 0.36 2.1 ± 0.44 2.2 ± 0.46 0.50 ± 0.06 0.18 ± 0.03° a‐T 3.1 ± 0.06 1.26 ± 0.51 2.2 ± 0.32*•♦ 2.2 ± 0.25 0.35 ± 0.04 0.19 ± 0.04°

Additional evidence that the rat renal interstitium contracts in vivo.

We recently provided highly suggestive preliminary evidence that the renal interstitium contracts reactively in vivo. We demonstrated that renal medullary direct interstitial volume expansion (rmDIVE = 100 μl bolus infusion of 0.9% saline (SS)/30 s) brought about a biphasic renal interstitial hydrostatic pressure (RIHP) response which was abolished when dibutyryl-cAMP was concomitant and interstitially infused. To assess more deeply the feasibility of the concept that the renal interstitium contracts in vivo, two experimental series (S1, S2) were performed in hydropenic rats subjected to acute left renal-denervation, hormonal clamping, and control of renal arterial pressure. In S1, RIHP and renal outer medullary blood flow (RoMBF) were continuously measured before and after a sudden micro-bolus (5μl) injection, into the renal medullary interstitium, of SS containing α-trinositol (α-TNS, anti-inflammatory drug) to either two doses 2 or 4 mM (SS + 2 α-TNS and SS + 4 α-TNS groups). No overall differences between groups in either ΔRIHP or %ΔRoMBF time courses were found; however, in the SS + 2 α-TNS group the data were less scattered and the ΔRIHP time course tended to peak faster and then persisted there, so that, this α-TNS dose was selected for S2. In S2, RIHP and RoMBF were similarly measured in rats randomly assigned to three groups: the CTR group (sham time-control), SS group (SS alone), and SS + α-TNS group. The micro-bolus injection of SS alone (SS group) was unable to increase ΔRIHP. The group with no micro-bolus injection (CTR group) experienced a decrease in ΔRIHP. The micro-bolus injection of SS + 2 α-TNS was accompanied by a differential increase in ΔRIHP (vs. CTR and SS groups). These responses were not associated with differential changes among groups in %ΔRoMBF or hemodilution parameters. These results provide additional evidence that the renal interstitium contracts in vivo.

RBF Autoregulatory Dynamics in a Virtual Step Response

Autoregulation of renal blood flow (RBF) is sum of myogenic (fast) and tubuloglomerular feedback (TGF, slow) components and their interaction. Blood pressure ( BP) and RBF (Transonics) were sampled at 128 Hz and stored at 2 Hz in n=6 hydropenic rats for 1hr before, then 1hr during, non-hypertensive infusion of nNOS blocker, SMTC, targeted to augment the tonic influence of TGF. 20-30 mmHg BP forcing was done with aortic occluder. Time series were processed by method of moving averages to generate the autoregulatory response to a unit step in BP, depicted as the fraction that is compensated by RBF autoregulation after a given lag. Results: Autoregulaory systems were stationary (step responses varied little throughout a given 1 hr recording). SMTC intensified the autoregulatory response, led to superautoregulation in several animals, and caused transient oscillations to appear at the myogenic frequency. Figure: left- pooled mean(sem) for control and SMTC periods. Right- mean(sem) difference between control and SMTC periods, which is useful measure of SMTC because 1st and 2nd period responses were correlated. The finding is consistent with generic tendency for feedback systems to oscillate when gain exceeds a threshold, and with existing theory that increasing the tone of TGF will make the myogenic component more reactive. These data confirm that this relatively simple analytical approach can reveal information about dynamic autoregulation that is instructive.

Renal NMDA receptors independently stimulate proximal reabsorption and glomerular filtration

N-methyl-D-aspartate receptors (NMDA) are expressed in the kidney, where little is known of their functional role. Several series of micropuncture experiments were performed in hydropenic rats using the NMDA channel blocker, MK801, and the NMDA coagonist, L-glycine, to probe NMDA for effects on single-nephron glomerular filtration rate (SNGFR) and proximal reabsorption (J(prox)). During intravenous infusion of MK801 or L-glycine, Henle's loop was perfused to manipulate SNGFR via tubuloglomerular feedback (TGF), thereby facilitating analysis of glomerulotubular balance. To confirm local actions on the kidney, MK801 was delivered to the glomerulus by microperfusion past the macula densa and to the proximal tubule by microperfusion into the early S1 segment. By all measures, MK801 acted on the glomerulus to reduce SNGFR, and acted on the proximal tubule to suppress J(prox), while having no effect on the responsiveness of TGF. L-Glycine raised SNGFR, dampened the TGF response, and could not be proved to independently stimulate proximal reabsorption. NMDA exerts a tonic vasodilatory influence on the glomerulus and a proreabsorptive effect on the proximal tubule. These combined effects allow NMDA to modulate SNGFR with minimal impact on late proximal flow. The full effects of L-glycine infusion on proximal tubule and TGF response do not extrapolate from the response to NMDA blockade.

The effect of AVP-V2 receptor stimulation on local GFR in the rat kidney.

The effect of AVP-V2 receptor agonist desmopressin, dDAVP, its non-peptide antagonist OPC-31260 and vehicle infusion on glomerular filtration rate (GFR) in the outer, middle and inner cortex was studied in both hydropenic and water diuretic Inactin anaesthetized female Sprague-Dawley rats using the aprotinin method. Two subsequent GFR measurements were carried out in the same kidney by injection of 125I- and 131I-labelled aprotinin before and after i.v. infusion of dDAVP, OPC-31260 or the vehicle. Acute infusion of dDAVP in hydropenic rats increased total GFR by 14% relative to vehicle infusion, whereas in water diuretic rats it had no effect relative to vehicle. No significant changes in arterial pressure (Pa) or renal blood flow (RBF) were recorded. Infusion of OPC-31260 reduced total GFR by 11% compared with vehicle. These results are consistent with the findings that a presensitization of the vasculature by high plasma levels of AVP is necessary for the renal vascular effects mediated by the V2 or V2-like receptors to occur. The ratio between inner and outer cortex GFR remained unchanged from control to experimental condition as follows: dDAVP infusion in hydropenic rats, 0.504 vs. 0.494 in control; vehicle infusion in hydropenic rats, 0. 393 vs. 0.392; OPC-31260 infusion in hydropenic rats, 0.517 vs. 0. 523; dDAVP in water diuretic rats, 0.547 vs. 0.543; vehicle in water diuretic rats, 0.413 vs. 0.417. Thus no significant difference in the GFR response was observed between superficial and deep cortical layers of the rat kidney.

Proximal tubular function in adult rats treated neonatally with enalapril.

Neonatal treatment with angiotensin-converting enzyme (ACE) inhibitors or the angiotensin II type-1 receptor antagonist losartan in rats induces irreversible renal histological abnormalities, mainly papillary atrophy, in association with an impairment in urinary concentrating ability. The aim of the present study was to assess proximal tubular function in adult rats treated neonatally with enalapril. Male Wistar rats received daily, intraperitoneal injections of either enalapril (10 mg kg-1) or isotonic saline vehicle from 3 to 24 days of age. In 15-week-old, hydropenic rats we analysed: (i) proximal tubular iso-osmotic fluid reabsorption using the method of lithium clearance; and (ii) maximal tubular D-glucose reabsorption (TmG), under pentobarbital anaesthesia. The main findings were that neonatally enalapril-treated rats showed: (i) reductions in absolute (APRH2O) and fractional (FPRH2O) iso-osmotic fluid reabsorption in the proximal tubules (APRH2O: 0.50 +/- 0.02 vs. 0.64 +/- 0.03 mL min-1 g KW-1, P < 0.05; FPRH2O: 58 +/- 3 vs. 68 +/- 2%, P < 0.05); and (ii) a normal TmG. In addition, during baseline clearance measurements neonatally enalapril-treated rats showed increases in urine volume and fractional excretion rates of sodium and potassium, a reduction in urine osmolality, whereas glomerular filtration rate and effective renal plasma flow were unaltered. These results suggest that neonatal ACE inhibition produces an irreversible, but differentiated, abnormality in proximal tubular function. Thus, the development of a normal proximal tubular function in the rat seems to be dependent on an intact renin-angiotensin system, (RAS) neonatally.

Endogenous angiotensin II modulates rat proximal tubule transport with acute changes in extracellular volume.

In the present study, we examined whether the effect of endogenously produced angiotensin II on proximal tubule transport in the male Sprague-Dawley rat is regulated by acute changes in extracellular volume. We measured the magnitude of endogenous angiotensin II-mediated stimulation of transport by sequentially perfusing proximal tubules in vivo, first with an ultrafiltrate-like solution, then by reperfusion of the same tubule with an ultrafiltrate-like solution containing 10(-8) M losartan (angiotensin II receptor antagonist). During volume contraction, 10(-8) M losartan decreased volume reabsorption from 4.20 +/- 0.50 to 1.70 +/- 0.30 nl . mm-1 . min-1 (P < 0.05), a decrease of 58.0 +/- 7.0%. In contrast, after acute volume expansion, 10(-8) M losartan decreased volume reabsorption from 1.84 +/- 0.20 to 1.31 +/- 0.20 nl . mm-1 . min-1 (P < 0.05), a decrease of 29.6 +/- 9.0%. In hydropenic rats, addition of exogenous luminal angiotensin II had no effect on transport. However, in volume-expanded rats, addition of 10(-8) M angiotensin II increased volume reabsorption from 2.10 +/- 0.34 to 4. 38 +/- 0.59 nl . mm-1 . min-1 (P < 0.005). These data are consistent with endogenously produced angiotensin II augmenting proximal tubule transport to a greater degree during volume contraction than after volume expansion.

Asymmetry of tubuloglomerular feedback effector mechanism with respect to ambient tubular flow.

The contribution of preglomerular resistance changes to the tubuloglomerular feedback (TGF) effector response was evaluated by micropuncture in hydropenic rats. Studies were performed in free-flowing nephrons to compare the fractional compensation (C) for perturbations in late proximal flow (VLP) with associated changes in glomerular capillary hydrostatic pressure (PGC). VLP was monitored by a noninvasive optical technique, and PGC was monitored by direct capillary micropuncture. Data were employed in conjunction with a model of glomerular filtration to analyze the TGF effector mechanism. C varied with the applied perturbation (VH), showing a single peak near the natural operating point. In contrast, the effect of TGF on PGC was asymmetric about the operating point, such that PGC was sensitive only to large positive perturbations. The model predicts that changes in preglomerular resistance account for only a small fraction of the integrated TGF response to small disturbances in the ambient state of the nephron but account for a greater share of the response to larger increments in flow. The TGF response in hydropenic rats is mediated by both pre- and postglomerular vascular elements.

Extravascular protein in the renal medulla: analysis by two methods

Two methods have been used to test for the presence of extravascular protein in the interstitium of the renal inner medulla. First, ascending vasa recta (AVR) segments were perfused with buffer containing 5 g/dl of albumin. The hydraulic pressure in the perfused vessel was varied to control transmembrane volume flux (Jv) to the interstitium. Interpolation to the point of zero Jv was employed to estimate interstitial Starling forces in the hydropenic rat papilla. Analysis of those experiments predicts that interstitial protein concentration (Ci) is high. When AVR segments are filled with oil, the oil column spontaneously breaks up as fluid is secreted into the lumen from the papillary interstitium. To obtain a lower limit on Ci, isolated AVR segments (IAS) filled with oil were sampled to measure protein concentration in the secreted fluid. In hydropenic rats, protein concentration was 3.4 +/- 0.5 and 5.2 +/- 0.2 g/dl in IAS and adjacent free-flowing AVR, respectively (P < 0.01). In rats subjected to furosemide and saline diuresis, the values were nearly identical, 4.7 +/- 0.2 and 5.2 +/- 0.2 g/dl, respectively. These separate experimental approaches corroborate a high concentration of protein in the renal inner medullary interstitium.

Effects of an atrial natriuretic peptide receptor antagonist on glomerular hyperfiltration in diabetic rats.

Previous studies from this laboratory implicated atrial natriuretic peptide (ANP) as a possible mediator of glomerular hyperfiltration in diabetic rats. In this study, the potential of HS-142-1 (HS), a novel polysaccharide compound with ANP receptor antagonist properties, to ameliorate glomerular hyperfiltration in diabetic rats was assessed. Initially, it was confirmed that a bolus iv injection of HS blocked both diuretic and natriuretic responses to exogenous ANP in normal Munich-Wistar rats. The acute effects of HS in moderately hyperglycemic diabetic rats with glomerular hyperfiltration and hyperperfusion were then determined. In diabetic rats, HS (20 mg/kg bolus iv) lowered GFR by approximately 46% from hyperfiltering (1.86 +/- 0.06 mL/min) to normal (1.13 +/- 0.13 mL/min) levels, whereas GFR in vehicle-treated diabetic rats remained unchanged (1.92 +/- 0.08 mL/min to 1.77 +/- 0.09 mL/min). In nondiabetic euvolemic rats, GFR was reduced by 20% after HS, whereas GFR in vehicle-treated controls was unchanged. In contrast, HS had no effect on GFR in hydropenic rats. Effective RPF was not significantly reduced in either diabetic or normal euvolemic rats in response to HS; consequently, significant reductions in filtration fraction were observed in both groups. Urine flow and sodium excretion rates were significantly reduced after HS in both diabetic and nondiabetic groups but not in hydropenic or vehicle-treated groups. These data show that HS ameliorates glomerular hyperfiltration in diabetic rats, further supporting the hypothesis that intrarenal actions of natriuretic peptides contribute significantly to glomerular hyperfiltration in experimental diabetes mellitus.

Effect of acute changes in glomerular filtration rate on Na+/H+ exchange in rat renal cortex.

Studies were undertaken in Munich-Wistar rats to assess the influence of changes in filtered bicarbonate (FLHCO3), induced by changes in GFR, on Na+/H+ exchange activity in renal brush border membrane vesicles (BBMV). Whole-kidney and micropuncture measurements of GFR, FLHCO3, and whole-kidney and proximal tubule HCO3 reabsorption (APRHCO3) were coupled with BBMV measurements of H+ gradient-driven 22Na+ uptake in each animal studied. 22Na+ uptake was measured at three Na+ concentration gradients to allow calculation of Vmax and Km for Na+/H+ exchange. GFR was varied by studying animals under conditions of hydropenia, plasma repletion, and acute plasma expansion. The increase in GFR, FLHCO3, and APRHCO3 induced by plasma administration correlated directly with an increase in the Vmax for Na+/H+ exchange in BBMV. The Km for sodium was unaffected. In the plasma-expanded rats, the Vmax for Na+/H+ exchange was 22% greater than in the hydropenic rats (P less than 0.025) whereas APRHCO3 was 86% greater (P less than 0.001). These results indicate that increases in FLHCO3, induced by acute increases in GFR, stimulate Na+/H+ exchange activity in proximal tubular epithelium. This stimulation is a mechanism which can, in part, account for the delivery dependence of proximal bicarbonate reabsorption.

Role of macula densa in diuretics-induced renin release.

Diuretic therapy may enhance renin release by various mechanisms, principally contraction of extracellular fluid volume and its effects, including a fall in arterial pressure. Awake hydropenic or volume-expanded rats received diuretics (amiloride and hydrochlorothiazide) that are known inhibitors of NaCl transport beyond the macula densa; also the well-known Na(+)-K(+)-2 Cl- transport system inhibitor furosemide was administered. We also evaluated the effect of a dose of ethacrynic acid (a drug that shares the same mechanism of action as furosemide but is not diuretic in the rat). The direct action of the diuretics on renin-producing cells was examined in isolated glomeruli; a rise in renin release was observed with the calmodulin inhibitor trifluoperazine (10(-5) M). Renin release in intact hydropenic rats was not altered by diuretic therapy, but furosemide increased plasma renin activity in hydropenic as well as in volume-expanded rats. This demonstrates the importance of furosemide inhibition of transport in the macula densa for its renin secretory action. None of the diuretics (amiloride, hydrochlorothiazide, ethacrynic acid, or furosemide) elicited changes in renin release from glomeruli (10(-6) to 10(-3) M); amiloride and hydrochlorothiazide (10(-4) to 10(-3) M) did not change renin release from slices, but 10(-3) M ethacrynic acid and furosemide increased renin secretion in this preparation. This suggests that an effect on the macula densa is essential in loop diuretic-mediated renin release.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of antidiuretic hormone on lithium as a marker for proximal tubule delivery.

To determine whether acute changes in antidiuretic hormone (ADH) alter the ability of lithium to quantitatively reflect proximal delivery, fractional lithium excretion (CLi/CIn) and late proximal (TF/P)In determined by micropuncture were measured in sodium-loaded rats following administration of desamino-8-D-arginine vasopressin (dDAVP) at rates equivalent to approximately half-maximal (20 pg/min) and maximal (40 pg/min) plasma levels of endogenous hormone. dDAVP dissociated the linear correlation between CLi/CIn and 1/(TF/P)In (r = 0.05) usually observed in sodium-loaded rats. Proximal delivery determined by lithium clearance underestimated (P less than 0.001) values obtained by micropuncture. Amiloride restored the linear correlation between CLi/CIn and 1/(TF/P)In in sodium-loaded dDAVP-treated rats (r = 0.70) and quantitative estimates of proximal delivery by each technique became equivalent. Indomethacin also reduced (P less than 0.001) estimates of proximal delivery obtained by lithium techniques in sodium-loaded rats. Water loading in hydropenic rats restored concordance between estimates of proximal delivery obtained by lithium clearance and micropuncture methods. Thus changes in ADH markedly alter the ability of lithium to function as a quantitative marker for proximal delivery. Furthermore, ADH may increase lithium uptake at distal nephron sites by an amiloride-sensitive pathway.

Effect of small-solute gradients on transcapillary fluid movement in renal inner medulla.

Volume efflux from descending vasa recta (DVR) of hydropenic rats occurs despite a higher oncotic pressure than hydraulic pressure. To explain this, we previously proposed that transcapillary small-solute gradients exert an additional driving force for volume efflux. This hypothesis was tested by micropuncture of DVR at the base and tip of the exposed renal papilla of control hydropenic and furosemide-treated rats. The DVR plasma osmolality at the base, 573 +/- 40, rose to 1,011 +/- 118 mosmol/kg H2O at the tip in control animals but was 356 +/- 8 and 377 +/- 6, respectively in furosemide animals, demonstrating that the axial osmotic gradient was nearly eliminated. The DVR plasma protein concentration in control hydropenic rats was higher at the tip, 6.8 +/- 0.4 g/dl, than at the base, 5.2 g/dl (P less than 0.01), because of volume efflux from DVR between base and tip, but was 5.4 +/- 0.3 and 5.5 +/- 0.2 g/dl at the base and tip, respectively, in furosemide animals, indicating virtual abolition of volume efflux. In DVR at the tip in control animals, plasma protein concentration positively correlated with plasma osmolality (R = 0.72, P less than 0.02). These results support the hypothesis that transcapillary small-solute gradients influence transcapillary fluid movement in DVR.

Role and regulation of glycerophosphorylcholine in rat renal papilla.

Glycerophosphorylcholine (GPC)--an organic solute which is considered to be involved in cellular osmoregulation in the renal medulla--was determined by means of an enzymatic assay in various zones of the rat kidney and in papillary tubule suspensions. In antidiuresis, GPC content in cortex, outer medulla and papillary tip was 0.64, 14.6, and 108.9 mmol/kg fresh weight, respectively. Significant concentrations of GPC could not be detected in the urine or in the peripheral plasma. The sharp increase in GPC concentration from cortex to papillary tip was partially abolished by the induction of diuresis by either waterloading or furosemide. These manoeuvres, however, did not change cortical GPC content. Papillary tubule suspensions prepared from hydropenic rats contained only slightly less GPC per g protein than whole papillae from antidiuretic animals. Incubation of tubules over 120 min did not lead to a significant loss of GPC which is in accordance with the low activity of GPC degrading enzymes in papillary tissue. The results confirm the intracellular localization of GPC and provide further evidence that this substance plays a substantial role in the osmoregulation of renal papillary cells.

Diuretic and natriuretic properties of prestegane B, a mammalian lignan.

The effect of increasing doses of prestegane B, a synthetic lignan, was examined in the anesthetized normal rat, using clearance methodology. Increasing doses of prestegane B 0.5, 1.0, 2.0, and 5.0 mg) were administered intravenously in our separate groups of hydropenic rats. Urine flow increased by 2.8 +/- 0.3, 4.5 +/- 0.5, 7.7 +/- 0.5, and 18.2 +/- 0.8 microliters/min, respectively, above control values. The rise of urinary sodium secretion was of similar magnitude and averaged 0.4 +/- 0.1, 0.8 +/- 0.2, 1.1 +/- 0.3, and 2.4 +/- 0.3 mu eq/min, respectively. No significant change in urinary phosphate excretion was obtained in all groups of rats, and glomerular filtration rate remained constant from control to experimental clearance periods. The natriuretic effect of prestegane B observed in this in vivo model could be related to the inhibition of the Na+-K+-adenosine triphosphate activity demonstrated in vitro in previous studies from our laboratory. The action of this substance is likely to be situated beyond the proximal tubule, since urinary phosphate was not altered. Prestegane B mimics the effects of other endogenous diuretic and natriuretic hormones, but its site of action and its effect on renal hemodynamics are obviously different.

Tubuloglomerular feedback control of distal fluid delivery: effect of extracellular volume.

A closed-feedback loop micropuncture method was used to examine tubuloglomerular feedback (TGF) and the regulation of distal fluid delivery in hydropenic rats (CON), moderately hemorrhaged rats (HEM), and rats given desoxycorticosterone (DOC) and 0.6% saline to drink. Distal delivery and TGF response curves were measured with four samples per nephron: a spontaneous early distal collection, two distal collections during moderate (7.5 nl/min) and saturating (30 nl/min) perturbations in nephron fluid load, and a proximal collection to measure single-nephron glomerular filtration rate (SNGFR) during TGF inhibition. Arterial pressure, predistal volume reabsorption, SNGFR, and early distal flow were significantly higher in DOC than in HEM; the CON group exhibited intermediate values. Except for a greater maximum TGF response in HEM, the normalized TGF responses were similar in all three groups, as was the regulation of distal fluid delivery. However, the TGF onset threshold and the TGF operating point, defined by the spontaneous rates of early distal flow and SNGFR, were reset such that distal fluid delivery and SNGFR were higher in DOC than in HEM, as was renal sodium excretion. The results show that the level around which TGF stabilizes distal fluid delivery is reset when extracellular fluid volume is altered.

Effect of atrial natriuretic factor in rat pregnancy.

Protocols were designed to 1) identify atrial natriuretic factor (ANF) bioactivity in pregnant rats, 2) compare ANF activity in atria from gravid and virgin Sprague-Dawley animals, and 3) assess renal and vascular responsiveness to ANF during gestation. Similar quantities of ANF were extracted from atria of term gravid and age-matched controls (assayed as the natriuretic response in awake virgins). We then purified by high-performance liquid chromatography a peptide identical to the synthetic 23 amino acid atriopeptin II (AP II). With equimolar injections of ANF, gravid rats had a blunted natriuretic response (P less than 0.05). However, when dose was based on estimated extracellular volume, awake virgin and pregnant animals displayed similar highly significant dose-response curves to infused AP II (R's 0.8). When hydropenic rats were assayed, both gravid and virgins required 10 times more ANF to initiate a diuresis; dose-response curves again being similar and highly significant. Decrements in blood pressure were also similar in pregnant and nongravid animals. Thus, despite an increased intravascular volume of 70% during gestation, cardiac content of ANF and the renal and vascular responsiveness to this peptide are similar to those in virgin rats, suggesting resetting of ANF-volume relationships during pregnancy.

Role of volume depletion in the glycerol model of acute renal failure.

Previous studies have demonstrated significant volume depletion early in the course of glycerol-induced acute renal failure. In addition, it has been shown that acute volume expansion within 6 h of glycerol administration reverses the fall in inulin clearance but volume expansion 12-24 h after glycerol fails to restore that parameter to normal. The current studies were performed to determine whether chronic volume expansion would prevent the fall in inulin clearance normally observed 18-20 h after the insult. Inulin clearance was therefore compared 18-20 h after glycerol in hydropenic rats and in rats undergoing chronic volume expansion by the constant infusion of Ringer solution at a rate of 5 ml/h initiated at the time of the glycerol injection. Inulin clearance was well maintained in the latter group, averaging 1.77 compared with 0.22 ml/min in the hydropenic group. Renal histology revealed no difference in the degree of tubular necrosis between groups but did show a marked decrease in cast formation in the animals undergoing chronic volume expansion. To determine the possible significance of these casts, micropuncture studies were carried out to measure proximal tubular pressures. Under hydropenic conditions these pressures were not different control animals and rats receiving glycerol when studied 18-20 h after glycerol. Following acute volume expansion, however, glycerol-treated rats demonstrated significantly higher pressures than control rats. On the other hand, proximal tubular pressures were comparable in glycerol-treated and control animals receiving the chronic volume expansion.(ABSTRACT TRUNCATED AT 250 WORDS)