The adrenal and vascular effects of angiotensin II and III in sodium depleted rats

The effects of angiotensin II and angiotensin III on mean arterial pressure, serum aldosterone, and serum corticosterone were studied in normal and sodium depleted, conscious rats. In normal rats, angiotensin III was 76% (p > 0.10) as potent as angiotensin II on aldosterone release but only 31% (p < 0.001) as potent on blood pressure. Following sodium depletion, the pressor responses to both angiotensin II and III were reduced (p < 0.001) (65% and 86% respectively). In addition, the release of aldosterone by both peptides was potentiated by sodium depletion as indicated by an increase in the slope of the dose-response curves. However, in the sodium depleted rats, angiotensin III was only 20% (p < 0.001) as potent as angiotensin II in stimulating aldosterone release. Small changes in serum corticosterone were noted following infusions of both peptides, but unlike the case with aldosterone, sodium depletion did not alter the serum corticosterone responses to the peptides. These $\text{in}$$\text{vivo}$ experiments taken with $\text{in}$$\text{vitro}$ studies support the interpretation that angiotensin III could function to control aldosterone release in altered sodium states either as a circulating hormone if present in concentrations far in excess of those of angiotensin II or as a local hormone formed in the adrenal from angiotensin II.     

Increased acridine orange uptake and lysosomal enzyme levels in rat liver after steroid administration

A steroid which stabilizes lysosomes $\text{in vitro}$ and a pyrogenic steroid which labilizes lysosomes $\text{in vitro}$ were compared with respect to their ability to modify lysosomal uptake and lysosomal enzyme levels $\text{in vivo}$. Cortisone acetate increased the uptake of acridine orange by rat liver lysosomes when the dye was administered by intrathoracic injection. The steroid increased and accelerated the uptake of acridine orange so that, in liver lysosomes from treated rats, the maximum uptake was double that of controls and was reached at 2h, whereas in controls the maximum uptake was at 4h after the injection of the dye. This large elevation of uptake is specific to the lysosomal fraction and is not seen in other subcellular fractions of rat liver. The specific activities of a lysosomal enzyme β-N-acetylglucosaminidase were increased in lysosomal fractions from cortisone acetate-treated rats. Etiocholanolone, a steroid which labilizes lysosome $\text{in vitro}$, similarly accelerated and increased acridine orange uptake by lysosomes but had little effect on lysosomal β-N-acetylglucosaminidase levels. Thus the ability of steroids to stabilize or labilize lysosomes $\text{in vitro}$ does not correlate with their effect on lysosomal uptake of injected substances $\text{in vivo}$, or with their ability to induce increased specific activities of lysosomal enzymes.     

Trilostane,an orally active inhibitor of steroid biosynthesis

Trilostane is a competitive inhibitor of 3β-hydroxysteroid dehydrogenase. $\text{In}$$\text{vitro}$, the drug inhibits conversion of pregnenolone to progesterone but does not alter conversion of cholesterol to pregnenolone nor progesterone to corticoid hormones. When given orally to rats, trilostane inhibits corticosterone and aldosterone production and elevates circulating levels of pregnenolone at doses lower than those that produce adrenal hypertrophy or inhibit gonadal steroidogenesis.     

Inhibitory action of norepinephrine on the induction of tryptophan-2,3-dioxygenase by cortisol: mediation by the alpha-receptor.

Induction of hepatic tryptophan-2,3-dioxygenase in rats by cortisol or corticosterone was inhibited on treatment with norepinephrine. The β-adrenergic blockers showed a small potentiating effect of the norepinephrine-mediated inhibition. The α-adrenergic blockers significantly reversed this inhibition, suggesting that norepinephrine acts $\text{via}$ the α-receptor in inhibition of the cortisol-mediated induction of this enzyme.     

Effects of naloxone on the secretion of prolactin and corticosterone induced by 5-hydroxytryptophan and a serotonergic agonist, mCPP

The effects of naloxone, an opiate “pure” receptor antagonist, on the release of prolactin and corticosterone in the rat were studied following the administration of the serotonin precursor 5-hydroxytryptophan or the serotonin receptor agonist (?) -$\text{m}$-chloropnehylpiperazine. Naloxone clearly antagonizes the release of prolactin induced by 5-hydroxytryptophan administered alone at a dosage of 50 mg/Kg/b.wt. or at dosage of 30 mg/Kg/b.wt. preceded 60 minutes before injection by the administration of the serotonin uptake blocker fluoxetine. The opiate antagonist does not modify the increase in blood level of prolactin induced by (?) ?$\text{m}$-chlorohenylpiperazine. Naloxone itself does not reduce the increase in plasma level of corticosterone induced by 5-hydroxytryptophan, 5-hydroxytryptophan +fluoxetine or (?)?$\text{m}$-chlorophenylpiperazine.The results suggest that endogenous opioids may be involved in the increase in serum level of prolactin induced by 5-hydroxytryptophan and also indicate the existence of different serotonergic neurotransmitter circuits capable of modulating the release of prolactin and corticosterone. A mutual interplay between serotonergic and opiate neurons may be involved in controlling the release of prolactin, but such an interplay does not seem to occur in the secretion of corticotrophin-releasing hormone.     

Effect of neonatally injected ACTH and ACTH analogues on eye-opening of the rat.

Three-day old rats were injected subcutaneously either with natural purified pig ACTH (ACTH 1–39), ACTH 1–24, or ACTH analogues as long-acting zinc-phosphate preparations. ACTH 1–39, ACTH 1–24, ACTH 1–18, ACTH 1–16 accelerated the time of eye-opening, whereas an extract of corticosteroids produced $\text{in}$$\text{vitro}$ by excised adrenals of ACTH-treated three-day old rats was ineffective. Injections with ACTH on the twelfth day of life had no effect on eye-opening. It is concluded that a neonatal injection with ACTH or closely related analogues with markedly less corticotropic activity can accelerate the time of eye-opening. This effect is not mediated by the adrenal cortex. The sensitive period for it appears to be shortly after birth.     

Synchronization of estrus in cyclic beef heifers with the prostaglandin analog alfaprostol

Seventy-eight Hereford-Angus crossbred heifers were injected intramuscularly twice with 6 mg of alfaprostol^b in 6 ml of propylene glycol. On each representative day of a 20-day estrous cycle (estrus = Day 0), either three or four heifers received their first injection. The second injection was given 12 days after the first, regardless of the response to the first injection. Thirty-nine heifers were not treated. The first alfaprostol injection reduced serum progesterone to less than 1 ng/ml in all heifers injected after Day 4. A total of 79.5% ($\text{62}\text{78}$) of the heifers exhibited estrus by five days after the first injection. Average interval from injection to estrus was 63 hours. The second injection occurred on Days 6 through 16 for all but one heifer, with 75.6% ($\text{59}\text{78}$) falling on Days 8 through 11 of the estrous cycle. Estrus was detected in 93.6% ($\text{73}\text{78}$) of the heifers within five days after the second injection, with an average interval to estrus of 66 hours.Day of cycle at second injection did not affect the interval to estrus. Conception occurred in 79.4% ($\text{58}\text{73}$) of the heifers inseminated in the five days after the second injection. Occurrence of estrus and conception was no different in treated heifers after five days of the insemination period than in nontreated heifers after 21 days of the insemination period, where 94.9% ($\text{37}\text{39}$) were observed in estrus and 83.8% ($\text{31}\text{37}$) conceived. Overall percent conception for a 55-day insemination period was 89.7 ($\text{70}\text{78}$) for treated and 87.2 ($\text{34}\text{39}$) for nontreated heifers. Day of cycle at first or second injection did not affect conception after the second injection. Some signs of estrus were observed in 11 of the 16 heifers injected before Day 5.A second trial to determine if alfaprostol induced luteolysis early in the cycle was conducted. Twenty purebred Angus, Hereford, or Simmental heifers received either one or two injections of alfaprostol on either Day 1, 2, 3, or 4. Only five heifers showed any signs of estrus, and the three that were inseminated did not conceive. Subsequent cycle length indicated that luteolysis occurred in only one heifer.Data suggest that alfaprostol is an effective luteolytic agent in cyclic beef heifers after Day 4 and that two injections 12 days apart will effectively synchronize estrus in heifers when distributed throughout the cycle at the first injection without affecting conception rate.     

Gonadotrophin-induced reduction in the steroidogenic responsiveness of the immature rat testis.

Injection of immature male rats with 10 IU human chorionic gonadotrophin resulted in a decrease in the steroidogenic capacity of the testis $\text{in vitro}$; this effect was apparent at 20 and 40 hours, but not at 65 hours after injection, and was not attributable to the loss of gonadotrophin receptors that occurs. This reduction in steroid output was strictly local as intra-testicular injection of gonadotrophin affected only the treated testis. It is concluded that the rat testis is capable of actively regulating its responsiveness to repeated hormonal stimulation.     

Stimulation of leydig cell testoterone secretion in vitro and in vivo in hypophysectomized rats by an agonist of luteinizing hormone releasing hormone

Injection of a luteinizing hormone-releasing hormone (LHRH) agonist into 55-day-old male rats which had been hypophysectomized 3 days earlier resulted in a 10- to 30-fold increase in the levels of testosterone in serum and testicular interstitial fluid (IF) in the 4h following injection. The levels achieved were within or above the normal range for intact untreated rats of this age. In similar animals, injection of LHRH agonist also enhanced the serum testosterone response to injected hCG at $\text{1}\text{1}\text{2}\text{h}$, but not at later times after injection, and by 24h reduced IF levels of testosterone suggested that LHRH agonist had begun to inhibit stimulation by hCG. $\text{In vitro}$, dispersed Leydig cells from untreated hypophysectomized rats showed a 2-fold increase in testosterone responsiveness to LHRH agonist when compared to cells from intact rats, and this change was associated with an 80% increase in the number of Leydig cell LHRH-receptors.     

The participation of corticosterone in luteinizing hormone releasing hormone (LH-RH) action on luteinizing hormone (LH) release from anterior pituitary cells in vitro

Corticosterone alone was not able to stimulate release of luteinizing hormone (LH) from anterior pituitary cells $\text{in}$$\text{vitro}$, but corticosterone in combination with luteinizing hormone releasing hormone (LHRH) augmented the release of LH into the culture media. These results may indicate that corticosterone may have the capacity to activate membrane receptors for LHRH in the gonadotrophs.     

Daily rhythms of glycogen synthetase and phosphorylase activities in rat liver: Influences of food and light

In rats fed during the night time (from 8 p.m. to 8 a.m.) the activities of liver glycogen synthetase $\text{I}$ and phosphorylase $\text{a}$ varied rhythmically during a 24 hour period. There was an inverse relationship between their levels; the level of synthetase $\text{I}$ rose to a maximum at around 6 a.m. and that of phosphorylase $\text{a}$ attained the peak value at around 6 p.m. Eye enucleation of rats did not affect significantly the daily rhythms of the enzymes. However, when food was offered only during the day time, the phases of both enzyme rhythms were shifted by about 12 hours. On starvation for 24 hours, the glycogen level was reduced almost to nil, but the daily rhythms of the enzymes were retained. It is thus very likely that the daily variations of the enzyme activities are not merely a passive effect of food intake, and that food can be a synchronizer or zeitgeber which sets up the characteristic rhythms of glycogen metabolism in the liver.     

Effects of hypophysectomy, adrenalectomy and corticosterone treatment on uptake and release of putative central neurotransmitters by rat hypothalamic tissue in vitro.

Uptake and release of radiolabeled serotonin, noradrenaline, dopamine, acetylcholine and GABA by rat hypothalamic tissue $\text{in virto}$ were examined following various treatments, which cause drastic changes in the tissue levels of corticosterone. Hypophysectomy affected both uptake and release of most of the neurotransmitters studied. However, adrenalectomy had a more selective effect, changing these processes for serotonin only. The uptake of radiolabeled serotonin by synaptosomes was decreased by about 30%, while its release from tissue slices upon depolarization with 40 mM K⁺ was increased 25%. Both of these changes could be prevented by injecting adrenalectomized rats with corticosterone.It is suggested that corticosteroid hormones might play a modulatory role in maintaining a certain functional activity level in central serotonergic neurons.     

Effects of thyrotropin-releasing hormone on behavioral and hormonal changes induced by beta-endorphin.

Adult male rats were injected intraventricularly either with saline or TRH (10 μg) 5 min prior to a second injection of either saline or β-endorphin (50 μg). The tripeptide produced a 100% increase of motility counts recorded over a 15 min period following the last injection, whereas β-endorphin decreased general motor activity. TRH pretreatment completely abolished the depressant effect of β-endorphin. In addition, TRH enhanced the PRL secretion induced by β-endorphin and antagonized the slight elevation of plasma GH levels observed in β-endorphin-treated rats. These results do not seem to be related to an interaction of TRH with opiate receptors since the tripeptide (10^?8, 10^?6 M) added $\text{in vitro}$ to rat brain homogenates did not alter the specific binding of ³H-naloxone nor affect the displacement by β-endorphin of such binding.     

Decreased liver cytosol glucocorticoid receptors in protein-deficient rats

A low protein diet (5% as compared to a control 21% protein diet, $\text{ad libitum}$) caused a significant decrease in the concentration of liver cytoplasmic glucocorticoid receptors; the equilibrium dissociation constant (Kd) did not change. The maximum decrease occurred in two weeks and was reversible upon substitution of the low protein by a control diet. This influence of protein deficiency could not be attributed to elevated plasma corticosterone levels since a comparable increase in plasma corticosterone of calorie-deficient rats (21% protein diet in restricted quantity) did not decrease glucocorticoid receptors and the difference in receptor levels of control and protein deficient animals persisted following adrenalectomy. These results suggest that glucocorticoids might not exert their usual biologic effects in the presence of protein malnutrition.     

The 20,000-dalton structural variant of human growth hormone: lack of some early insulin-like effects

In contrast to the major form of human growth hormone the 20,000-dalton (20K) variant of the hormone produced no decrease in either serum glucose of free fatty acids one hour after injection into fasted, hypophysectomized rats. Furthermore, the variant caused no rise in serum free fatty acids after 5 hours. $\text{In}\text{vitro}$ experiments utilizing epididymal adipose tissue from hypophysectomized rats indicated that 20K was unable to accelerate glucose utilization as measured by glucose uptake and CO₂ formation. The data show that this form, even though growth promoting, lacks some of the metabolic properties attributed to growth hormone. We conclude that an insulin-like effect is not necessarily a prerequisite for growth promoting activity.     

Rapid pulsatile corticosterone secretion in the rat is not confirmed

We attempted to confirm and extend a previous suggestion by other workers that, in the rat, corticosterone may be released as a series of very short pulses with a period of one minute. We measured the corticosterone concentration in the blood of chronically cannulated, unanaesthetised male rats, repeatedly, at ten second intervals, for periods of up to 25 minutes while the rats were engaged in normal activity or sleep or were subject to acute or chronic stress. We could find $\text{no}$ evidence of the proposed rapid pulsatile secretion and suggest that the earlier finding may have been artifactual.     

Mechanism of corticotropin action in rat adrenal cells I. The effects of inhibitors of protein synthesis and of microfilament formation on corticosterone synthesis

The contribution of protein synthesis and formation of microtubules and microfilaments to corticotropin-stimulated steriodogenesis in rat adrenal cell suspensions has been assessed by use of a series of inhibitors to each function. Five inhibitors of protein synthesis (cycloheximide, puromycin, blastocidin S, anisomycin, and trichodermin) each exhibited time-dependent inhibition of corticotropin-stimulated steroidogenesis. For the first 30 min, steroidegenesis was more extensively inhibited than protein synthesis, after which the effectiveness of the inhibitors diminished on steroidegenesis but not on protein synthesis. The reversal effects was not observed at high levels of inhibitors. One inhibitor of microfilament fromation (cytochalasin B) and four inhinitors of microtubule formation (colchicine, podophyllotoxin, vinblastine sulfate and griseofulvin) inhibited steroidogenesis without inhibiting protein synthesis and without any reversal effect with prolonged incubation. The actions of all ten inhinitors were shown to be fully reversible. Cell superfusion of adrenal cells showed that the decay of steroidogenesis upon addition of all the protein synthesis inhibitors was similar to decay upon removal of corticotropin from the medium ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{= 4–6}\text{min}$). Recoveries from inhibition upon removal of the inhibitors were similar to each other and comparable to initial corticotropin stimulation of the cells (lag of 3–5 min, $\text{f}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{= 7–9}\text{min}$). Similar kinetics of inhibition and recovery were observed for vinblastine sulfate while a direct inhbition of cytochrome P-450_sec by an aminoglutethimide was complete within 1 min and was rapidly reversed.Injection of each inhibitors (all classes) into hypophysectomized rats inhibited the elevation of plasma corticosterone by corticotropin. The extent of cholesterol combination with cytochrome P-450_sec in adrenal mitochondria isolated from these rats was also decreased by all inhbitors. Decreases in plasma corticosterone correlated directly with decreases in cholesterol combination with cytochrome P-450_sec (r = 0.94).It is concluded that protein synthesis and steroidogenesis must be intimately coupled propbably due to the requirement of a labile protein for cholesterol transport to cytochrome P-450_sec. An involvement of microtubules and microfilaments in this process is clearly indicated.     

Anti‐inflammatory effects of melatonin in a rat model of caerulein‐induced acute pancreatitis

The purpose of our study was to evaluate the protective effect of melatonin in a rat model of caerulein‐induced acute pancreatitis. For the induction of experimental acute pancreatitis, four subcutaneous injections of caerulein (20 µg kg^–1 body weight) were given to Wistar rats at 2‐h intervals. Melatonin was injected intraperitoneally (25 mg kg^–1 body weight) 30 min before each caerulein injection. After 12 h, rats were sacrificed by decapitation. Blood and pancreas samples were collected and processed for serological and histopathological studies, respectively. Lipase, α‐amylase, corticosterone, total antioxidant power and cytokines interleukin (IL)‐1β, IL‐4 and tumour necrosis factor (TNF)‐α were determined using commercial kits. ANOVA and Tukey tests (P < 0.05) were performed for the statistical analysis of the results. Results showed that the administration of melatonin reduced histological damage induced by caerulein treatment as well as the hyperamylasemia and hyperlipidemia. Corticosterone and antioxidant total power were also reverted to basal activities. Furthermore, melatonin pre‐treatment reduced pro‐inflammatory cytokines IL‐1β and TNF‐α and increased the serum levels of anti‐inflammatory cytokine IL‐4. In conclusion, the findings suggest that the protective effect of melatonin in caerulein‐induced acute pancreatitis is mediated by the anti‐inflammatory ability of this indolamine. Thus, melatonin may have a protective effect against acute pancreatitis. Copyright © 2013 John Wiley & Sons, Ltd.     

Action of Solanum malacoxylon on calcium metabolism in the rat

The administration of an aqueous extract of the leaves from $\text{Solanum malacoxylon}$ to vitamin D-deficient rats fed a normal calcium, normal phosphorus diet markedly increased serum calcium concentration within 48 hours. The $\text{Solanum malacoxylon}$ extract also stimulated intestinal calcium transport in the vitamin D-deficient rat but was without effect on the mobilization of calcium from bone. The extract from 100 mg of dry $\text{Solanum malacoxylon}$ leaves was more effective than 25 units of vitamin D given daily to vitamin D-deficient rats in stimulating intestinal calcium transport but its effect was not additive to that of the vitamin D. The results demonstrate that the action of $\text{Solanum malacoxylon}$ is independent of vitamin D and, although it can substitute for vitamin D in the stimulation of intestinal calcium transport activity, it cannot substitute for vitamin D in the mobilization of calcium from bone.     

Inhibition of the effect of lithium on brain inositol by atropine and scopolamine.

Atropine and scopolamine were found to inhibit the reduction in levels of brain $\text{myo}$-inositol (inositol) which occurs in lithium treated rats. There was no effect on the elevation of serum inositol levels induced by lithium. The anticholinergic agents produced no changes in inositol levels in brain or serum when administered alone and also they did not affect the concentrations of lithium in the tissues. The effect of lithium on brain inositol was not inhibited by methylatropine or methylscopolamine.