Radioimmunoassay for human growth hormone-releasing factor (hGRF 1-40): comparison of plasma immunoreactive GRF after intravenous and subcutaneous administration to rats

A homologous radioimmunoassay (RIA) system was developed for human GRF 1-40 and used to measure immunoreactive (IR) concentrations of the peptide in rats to determine some of its pharmacokinetic characteristics after intravenous (i.v.) and subcutaneous (s.c.) administration. A plot of the disappearance of IR-hGRF from plasma after a single intravenous injection was fitted by a biexponential curve, analysis of which gave a half-life of 3.2 +/- 0.2 min for the initial distribution phase and 57.3 +/- 1.5 min for the elimination phase. Comparison of areas under the plasma IR-hGRF/time curves after injection of identical doses of hGRF 1-40 showed that the amount detected in the circulation after it was injected s.c. was only 14-16% of the amount detected after i.v. administration. Such results may indicate degradation of a substantial proportion of the dose of the peptide at the site of injection or during its transfer to plasma; this should be borne in mind when undertaking s.c. administration for clinical purposes or in assessing the effect of GRF analogues.     

Growth hormone-releasing factor analogue (hGRF1-29NH2): immunoreactive-GRF plasma levels after intravenous and subcutaneous administration

A homologous radioimmunoassay (RIA) system for human growth hormone-releasing factor 1-29NH2 (hGRF 1-29-NH2) was developed and applied to the measurement of immunoreactive (IR) concentrations of the peptide in anaesthetized rats to determine some of its pharmaco-kinetics after i.v. and s.c. administration. Analysis of the biphasic disappearance curve of IR-hGRF-1-29NH2 from plasma after i.v. injection (10 micrograms) gave values for the half-lives of the initial distribution phase (alpha) and for the elimination phase (beta) of 1.9 +/- 0.2 min and 10.4 +/- 0.2 min respectively. There was rapid uptake of the peptide from the site of s.c. injection but comparison of areas under the plasma IR-hGRF1-29NH2/time curves showed that the estimated total amount in the circulation after s.c. injection was only 4% of that obtained after i.v. administration. A large degree of degradation of the peptide is indicated either at the site of injection or during transfer to plasma; this susceptibility to rapid breakdown is reflected in the short half-life of the peptide in the circulation. Therefore the measurement of the above parameters is a prerequisite when assessing potency of GRF analogues in-vivo and when implementing suitable dosage regimens for clinical purposes.     

PLASMA IMMUNOREACTIVE GHRH AND SERUM GH CONCENTRATIONS FOLLOWING PULSATILE GHRH 1-40 ADMINISTRATION IN GH DEFICIENT CHILDREN

We have studied the clearance from plasma of immunoreactive growth hormone releasing hormone 1-40 (IR-GHRH) following intravenous (i.v.) and subcutaneous (s.c.) administration and the relationship between exogenous plasma IR-GHRH concentrations and GH secretion in five GH insufficient children receiving long term nocturnal pulsatile GHRH 1-40. The i.v. studies with GHRH 1-40 1 micrograms/kg demonstrated a distribution half life (t1/2) of 3.9 (SD 0.9) min and an elimination t1/2 of 53.1 (SD 3.2) min. In the s.c. studies the elimination phase was similar to the i.v. results but the transit time to the GHRH peak was slower than the i.v. distribution t1/2 9.9 (SD 3.6) min. These characteristics were maintained during successive pulses of subcutaneous GHRH. The mean IR-GHRH peaks following s.c. GHRH 1-40 administration of 1 microgram/kg and 2 micrograms/kg were 37- and 18-fold lower respectively than the mean IR-GHRH peak observed after the i.v. 1 microgram/kg bolus study. A significant correlation was shown between peak plasma IR-GHRH and serum GH concentrations during the s.c. (r = 0.75) but not the i.v. studies. Pulsatile GHRH administration has been shown to stimulate GH secretion and growth acceleration in GH insufficient children. Knowledge of the relationship between GHRH 1-40 absorption from the subcutaneous site and GH secretion is important for the development of an optimal GHRH treatment regimen in GH insufficient children.     

Plasma concentrations of cholecystokinin octapeptide and food intake in male rats treated with cholecystokinin octapeptide

Intraperitoneal injection of 5 micrograms cholecystokinin octapeptide (CCK-8) into male rats deprived of food for 48 h produced a transient (less than 15 min) increase in plasma levels of CCK-8 but suppressed food intake for an extended period (45 min). Plasma concentrations of CCK-8 after i.p. injection of CCK-8 were raised to levels which were fairly comparable to those after feeding. Intracerebroventricular (i.c.v.) injection of the CCK antagonist proglumide (100 micrograms) reversed the effect of CCK-8 on food intake, while i.p. injection of proglumide (100 micrograms) did not have this effect. Feeding increased the plasma concentrations of somatostatin and gastrin but not of oxytocin, and somatostatin and oxytocin but not gastrin were released in response to i.p. injection of CCK-8. However, neither somatostatin nor oxytocin affected food intake, and their release in response to CCK-8 was unaffected by i.c.v. injection of proglumide. These results support the suggestion that CCK-8 is a physiological 'satiety' peptide, which can affect food intake in rats by mechanisms involving both peripheral and central CCK receptors.     

The effect of systemic administration of dopamine and apomorphine on Plasma LH and prolactin concentrations in conscious rats.

The effect of systemic administration of various doses of dopamine (DA) and apomorphine (APM) on plasma gonadotropin and prolactin (Prl) concentrations in ovariectomized (OVX) as well as in ovariectomized, estrogen-progesterone (OEP)-primed rats bearing indwelling jugular venous catheters was evaluated. Intravenous (i.v.) infusion or pulse injection of 0.9% NaCl had no significant effect on plasma titers of LH or Prl. I.v. infusion of DA at 4 micrograms/kg-b.w./min induced a progressive increase in circulating LH concentration in OEP rats while infusion at a similar dose in OVX animals had no effect on plasma LH. I.v. injection of 100 micrograms DA or APM significantly increased LH at 15 min in OVX rats. Similarly, in OEP rats 100 micrograms of DA elevated plasma LH at 30 and 90 min while APM induced a significant elevation of plasma LH at 15 min after injection. In OVX rats injection of DA i.p. at a dose of 5 mg/kg-b.w. did not alter plasma LH levels, but a dose of 50 mg/kg-b.w. produced a significant reduction in plasma LH concentration. APM injected i.p. at either 5 or 50 mg/kg-b.w. doses was nearly equally effective in lowering plasma LH and the suppressive effect was significantly greater than with similar doses of DA. A single injection of LH-RH (100 ng in 0.2 ml of 0.9% NaCl) in animals pretreated 15 min earlier with an effective dose of APM (5 mg/kg-b.w.) produced a peak increase in LH titers 15 min after injection. The increment in plasma LH following LH-RH in APM-treated rats was comparable to that in rats which had received saline instead of APM. Prl levels were significantly lowered by each dose of DA and APM in OVX as well as in OEP rats. There was no significant change in plasma FSH titers induced by either drug in any of the experiments. It is concluded that DA may have different actions depending upon the dose and the endocrine state of the animal. Thus, i.v. infusion of low doses of DA in OEP animals or by pulse injection in both OVX and OEP rats can elevate plasma LH by activating the release of LH-RH from the hypothalamus, while large doses of DA in OVX animals may suppress the release of LH-RH.     

Pharmacokinetics and pharmacodynamics of subcutaneously administered U40 and U100 formulations of regular human insulin.

Action profiles of 12 U of regular human insulin (Actrapid HM) administered subcutaneously as a U40 or U100 formulation were studied. Euglycaemic glucose clamps were performed on two separate days in 8 healthy subjects (basal i.v. insulin infusion 0.1 mU/kg/min, plasma glucose 5.0 mmol/l, mean +/- SD age 25 +/- 2 years, BMI 22.7 +/- 1.4 kg/m2). Serum insulin concentrations increased after injection of U40 or U100 from similar baseline values to maximal individual concentrations of 305 +/- 79 vs. 285 +/- 62 pmol/l (NS) after 90 +/- 33 vs. 114 +/- 58 min (NS). Ten, 15, and 20 min post injection insulin concentrations were significantly higher by an average of 30 pmol/l after U40 insulin vs. U100 insulin (p less than 0.05). Glucose infusion rates increased from comparable baseline rates to maximal individual glucose infusion rates of 10.7 +/- 2.4 vs. 10.9 +/- 3.0 mg/kg/min (NS) after 172 +/- 51 vs. 169 +/- (39) min (NS). At the three time points when significantly different serum insulin concentrations occurred soon after insulin injection, glucose infusion rates were not significantly different between U40 and U100. Although small differences in insulin pharmacokinetics were detected early after s.c. insulin injection (U40 was absorbed faster than U100 insulin) the pharmacodynamics of the U40 and U100 formulation of regular human insulin appear to be comparable in healthy subjects.     

Pegfilgrastim,a sustained-duration form of filgrastim,significantly improves neutrophil recovery after autologous marrow transplantation in rhesus macaques

Daily administration of filgrastim decreases the duration of severe neutropenia in the clinical setting. A sustained-duration form of filgrastim, pegfilgrastim, significantly reduces scheduling protocols to a single injection per chemotherapy cycle while maintaining therapeutic efficiency. We examined the ability of a single injection of pegfilgrastim to significantly improve neutrophil recovery following autologous bone marrow transplantation (AuBMT) in rhesus macaques. On day 1, postmyeloablation (920 cGy x-irradiation) and AuBMT, animals received either 0.1% autologous serum for 18 consecutive days (n=13), or single doses of pegfilgrastim via the subcutaneous (s.c.) or intravenous (i.v.) route (300 or 100 micro g/kg), or a single dose of filgrastim at 300 micro g/kg via the s.c. or i.v. route, or filgrastim at 10 micro g/kg via the s.c. route (n=4) on a daily basis (range=days 12-17). Pharmacokinetic parameters and neutrophil recovery were assessed. A single dose of pegfilgrastim via the i.v. or s.c. route was as effective as daily filgrastim administration, resulting in significant improvement of neutrophil recovery after myeloablation and ABuMT. Effective pegfilgrastim plasma concentrations were maintained in neutropenic animals until after the onset of hematopoietic recovery. Enhanced pharmacokinetics in AuBMT cohorts are consistent with self-regulating, neutrophil-mediated clearance.     

Seasonal and steroid-dependent effects on the modulation of LH secretion in the ewe by intracerebroventricularly administered beta-endorphin or naloxone

The natural opioid ligand, beta-endorphin, and the opioid antagonist, naloxone, were administered intracerebroventricularly (i.c.v.) to evaluate effects on LH secretion in ovariectomized ewes and in ovariectomized ewes treated with oestradiol-17 beta plus progesterone either during the breeding season or the anoestrous season. Ovary-intact ewes were also studied during the follicular phase of the oestrous cycle. Jugular blood samples were taken at 10-min intervals for 8 h and either saline (20-50 microliters), 100 micrograms naloxone or 10 micrograms beta-endorphin were injected i.c.v. after 4 h. In addition, luteal phase ewes were injected i.c.v. with 25 micrograms beta-endorphin(1-27), a purported endogenous opioid antagonist. In ovariectomized ewes, irrespective of season, saline and naloxone did not affect LH secretion, but beta-endorphin decreased the plasma LH concentrations, by reducing LH pulse frequency. The effect of beta-endorphin was blocked by administering naloxone 30 min beforehand. Treating ovariectomized ewes with oestradiol-17 beta plus progesterone during the breeding season reduced plasma LH concentrations from 6-8 micrograms/l to less than 1 microgram/l. In these ewes, saline did not alter LH secretion, but naloxone increased LH pulse frequency and the plasma concentrations of LH within 15-20 min. During anoestrus, the combination of oestradiol-17 beta plus progesterone to ovariectomized ewes reduced the plasma LH concentrations from 3-5 micrograms/l to undetectable levels, and neither saline nor naloxone affected LH secretion. During the follicular phase of the oestrous cycle, naloxone enhanced LH pulse frequency, which resulted in increased plasma LH concentrations; saline had no effect.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative stimulation of growth hormone secretion in anaesthetized chickens by human pancreatic growth hormone-releasing factor (hpGRF) and thyrotrophin-releasing hormone (TRH)

Plasma concentrations of growth hormone (GH) were elevated in anaesthetized male domestic fowl following the intravenous administration of either synthetic human pancreatic GH-releasing factor 1-44 (NH2) (hpGRF) or synthetic thyrotrophin-releasing hormone (TRH). In 6-week-old chicks the plasma GH level was elevated between 5 and 10 min after the injection of hpGRF at doses between 1 and 80 micrograms/kg. The magnitude of the response increased with doses of hpGRF between 1 and 10 micrograms/kg but declined with higher doses. The GH concentration rapidly declined between 10 and 20 min and between 20 and 40 min after injection. The administration of TRH had similar effects on GH secretion, although the responses were greater than with comparable doses of hpGRF, and the most effective dose (1-1.4 micrograms/kg) was less than with hpGRF. In anaesthetized adult cockerels GH secretion was also increased by the administration of hpGRF (1-20 micrograms/kg) or TRH (0.1-80 micrograms/kg) and in both cases the dose-response relationship was biphasic. The maximal response to TRH in adult birds was again greater than that produced by hpGRF although the response was less than that elicited in immature birds and required a higher dose (20 micrograms/kg) of TRH. The optimal dose of hpGRF and the magnitude of the GH response induced in adult birds was comparable with that in immature chicks. These results demonstrate provocative effects of TRH and hpGRF on GH secretion in the domestic fowl. The sensitivity of the GH response to TRH suggests that it may have a physiological role in the hypothalamic control of GH secretion.     

Oral administration of growth hormone (GH)-releasing peptide stimulates GH secretion in normal men.

Intravenous infusions of the synthetic hexapeptide GH-releasing peptide (His-DTrp-Ala-Trp-DPhe-Lys-NH2; GHRP) specifically stimulate GH release in man. To determine whether orally administered GHRP stimulates GH secretion, 10 normal men received oral doses of placebo, 30, 100, and 300 micrograms/kg GHRP, and an iv injection of 1.0 micrograms/kg GHRP at weekly intervals in a single blind, randomized design. Serum GH concentrations were measured in blood samples obtained at 5-min intervals for 1 h (0700-0800 h) before and 4 h (0800-1200 h) after each dose. Mean (+/- SE) peak GH concentrations were 4.0 +/- 1.5, 5.2 +/- 1.6, 9.2 +/- 3.3, 18 +/- 3.7, and 26 +/- 5.6 micrograms/L for placebo; 30, 100, and 300 micrograms/kg oral GHRP; and 1 micrograms/kg iv GHRP, respectively; mean 4-h (0800-1200 h) integrated GH concentrations were 312 +/- 109, 406 +/- 159, 698 +/- 284, 1264 +/- 303, and 1443 +/- 298 min.micrograms/L, respectively. To analyze changes in the pulsatile pattern and amount of GH secretion after the administration of GHRP, a waveform-independent deconvolution method was used to estimate GH secretion rates. Variable increases in GH secretion after placebo and GHRP treatments were observed. Despite this variability, weighted least squares linear regression revealed that increasing doses of oral GHRP progressively stimulated GH secretion (P less than 0.005); similar relationships were observed for the peak GH concentration and 4-h integrated GH concentrations. The GH responses to oral GHRP (300 micrograms/kg) and iv GHRP (1 microgram/kg) were significantly greater than that to placebo (P less than 0.05) and were comparable in magnitude. Pairwise comparisons revealed that increases in GH concentrations and secretion rates after the 30 and 100 micrograms/kg oral doses of GHRP were not significantly different from those after placebo. The increase in GH secretion after GHRP treatment was accounted for entirely by an increase in the amplitude of GH secretory events, as no significant increase in the number of GH secretory pulses was observed. The onset and duration of action of GHRP were analyzed by a proportional hazards general linear regression model. Intravenous GHRP had a more rapid onset of action than all doses of oral GHRP (P less than 0.02). Increasing doses of oral GHRP resulted in earlier GH responses (P = 0.006). However, the duration of the GH response was similar for iv GHRP and all doses of oral GHRP, averaging 120-150 min.(ABSTRACT TRUNCATED AT 400 WORDS)     

Opioid modulation of FSH, growth hormone and prolactin secretion in the prepuberal gilt.

The role of endogenous opioid peptides (EOP) in modulating GH, prolactin (PRL) and FSH secretion was evaluated in prepuberal (P) gilts. In experiment I, P gilts received 1 (n = 2), 3 (n = 3) or 6 (n = 3) mg naloxone (NAL)/kg body weight i.v. Blood was collected every 15 min for 2 h prior to and 2 h after NAL and an additional 1 h after 100 micrograms gonadotrophin-releasing hormone (GnRH) i.v. In experiment II, P and mature (M) gilts were ovariectomized. Three weeks after ovariectomy, P and M gilts were injected twice a day for 10 days with either 0.85 mg progesterone (P4)/kg body weight or oil vehicle (V), resulting in the following groups: PP4 (n = 11), PV (n = 10), MP4 (n = 11) and MV (n = 10). All gilts received 1 mg NAL/kg body weight on the last day of treatment. Blood samples were collected every 15 min for 4 h before and 2 h after NAL and an additional 1 h after 100 micrograms GnRH i.v. In experiment III, six P and five M gilts were ovariectomized and surgically implanted with intracerebroventricular (i.c.v.) cannulae. Blood was collected every 15 min for 3 h before and 3 h after i.c.v. injection of 500 micrograms morphine in artificial cerebrospinal fluid (CSF) or 250 microliters CSF. In experiment I, all doses of NAL failed to alter PRL secretion, while NAL increased (P less than 0.05) GH secretion in three out of eight gilts. However, NAL suppressed (P less than 0.05) FSH concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Stimulation of in-vivo growth hormone secretion in young chickens by rat hypothalamic growth hormone-releasing factor and synthetic analogues

Rat hypothalamic GH-releasing factor (rhGRF), at doses between 0.1 and 10 micrograms/kg, increased plasma GH concentrations in immature domestic fowl 5-10 min after i.v. injection. Sodium pentobarbitone anaesthesia blunted the GH responses to rhGRF, although in both conscious and anaesthetized chicks the maximal responses were induced by a dose of 1 microgram rhGRF/kg. The stimulatory effect of rhGRF on in-vivo GH secretion was less than that provoked by corresponding doses of human pancreatic GRF, but greater than that elicited by two rhGRF analogues, (Nle27)-rhGRF(1-32) and (Nle27)-rhGRF(1-29). These results demonstrate that the chicken pituitary is responsive to mammalian GRF and provide evidence of structure-activity relationships of GRF in the domestic fowl.     

The impact on coagulation of an intravenous loading dose in addition to a subcutaneous regimen of low-molecular-weight heparin in the initial treatment of acute coronary syndromes

OBJECTIVES: We sought to quantify the impact of adding an intravenous loading dose to a subcutaneous regimen of enoxaparin in patients with an acute coronary syndrome (ACS). BACKGROUND: It is unclear whether an intravenous (i.v.) loading dose of enoxaparin should be added to a subcutaneous (s.c.) regimen in patients with ACS. METHODS: Patients admitted with ACS were randomized to i.v.+s.c.(n = 14) or s.c. alone (n = 11) enoxaparin treatment. Coagulation markers were measured at nine time points during the first 24 h of treatment. RESULTS: The i.v.+s.c. therapy immediately resulted in therapeutic anti-Xa levels, which remained significantly higher for 6 h compared with s.c. alone, without reaching excessively high levels. A rapid decrease of plasma prothrombin fragments 1+2 (F(1+2)) levels was observed as soon as 5 min after the i.v. injection (33% lower; p = 0.007), and these levels remained lower up to 2 h after the start of treatment compared with SQ alone. The ex vivo thrombin generation time was maximally prolonged at 5 min post-injection in the i.v.+s.c. group and remained significantly prolonged up to 6 h post-injection compared with s.c. alone. The tissue factor pathway inhibitor plasma activity was immediately increased by 194% with i.v.+s.c., whereas the maximum increase with s.c. alone was 47% at 3 h. CONCLUSIONS: Therapeutic plasma levels of enoxaparin are achieved significantly earlier by an i.v.+s.c. regimen compared with s.c. alone, without leading to unacceptably high levels. As the risk of thrombotic complications is greatest early after admission, the observed differences in antithrombotic effects may translate into a clinical benefit. However, this remains to be established.     

Thyrotrophin-releasing hormone-induced growth hormone (GH) secretion in anaesthetized chickens: inhibition by GH-releasing factor at central sites.

Intracerebroventricular (i.c.v.) administration of GH-releasing factor (GRF) (at 1 or 10 micrograms) to anaesthetized immature (6- to 8-weeks-old) or adult (greater than 24-weeks-old) domestic fowl had no effect on basal GH concentrations in peripheral plasma, but suppressed (after 20 min) the acute GH response to exogenous (i.v.) thyrotrophin-releasing hormone (TRH) (1 micrograms/kg). The i.c.v. injection of GRF also reduced the content of somatostatin (SRIF) and dopamine (DA) in the hypothalamus, while increasing the concentration of the DA metabolite 3,4-dihydroxyphenyl acetic acid (DOPAC) and the DOPAC/DA ratio. The release of SRIF from hypothalamic tissue was stimulated in vitro by 100 nmol GRF/l. The inhibitory effect of i.c.v. GRF on TRH-induced GH secretion was blocked when it was simultaneously injected i.c.v. with SRIF antiserum. These results demonstrate central effects of GRF on avian hypothalamic function and suggest an inhibitory role for this peptide in GH regulation, possibly mediated through increased SRIF secretion.     

Prolonged administration of human atrial natriuretic peptide in healthy men. Reduced aldosteronotropic effect of angiotensin II

The effect of angiotensin II (5, 10, 20 ng/kg/min) on blood pressure and on the plasma concentrations of aldosterone was studied in six healthy men with and without the concomitant administration of synthetic human atrial natriuretic peptide given 1) as an i.v. bolus of 25 micrograms followed by a 6-hour infusion of 25 micrograms/hr or 2) as an i.v. bolus of 175 micrograms followed by a 6-hour infusion of 100 micrograms/hr. The pressor effect of angiotensin II (i.e., the rise of mean blood pressure above individual basal levels) remained unchanged during the administration of both doses of human atrial natriuretic peptide. The angiotensin II-induced rise in plasma concentrations of aldosterone in terms of absolute values) was reduced by human atrial natriuretic peptide during both trials. The rise in plasma concentrations of aldosterone above individual basal concentrations was also reduced during the administration of human atrial natriuretic peptide, although this effect was only marginal during the low dose experiment. These effects of human atrial natriuretic peptide support the contention that its therapeutic impact in hypertensive patients might be mediated in part by a reduction of high aldosterone concentrations.     

Effects of neonatal treatment with monosodium glutamate on growth hormone release induced by clonidine and prostaglandin E1 in conscious male rats

Effects of the centrally acting alpha-adrenergic agonist, clonidine, on growth hormone (GH) secretion was studied in conscious male rats pretreated with monosodium glutamate (MSG) during the neonatal period. GH secretory profiles in individual adult rats were obtained by repeated blood samplings every 10-20 min from 10.00 to 17.00 h. GH secretion was pulsatile with mean peak values at around 12.40 and 15.20 h in control rats. When clonidine (15 micrograms/100 g body weight) was injected intravenously into control rats at 14.00 h in the interval between two anticipated spontaneous GH bursts, plasma GH was increased with a mean peak value 20 min after the injection, and the following anticipated spontaneous GH burst was not observed during the experiment. In the rats neonatally treated with MSG (4 mg/g body weight, s.c.), which causes selective destruction of the hypothalamic arcuate nucleus, plasma GH response to clonidine as well as the spontaneous GH bursts were considerably blunted, whereas prostaglandin E1 (5 micrograms/100 g body weight, i.v.) caused an abrupt increase in plasma GH levels in these animals. These results suggest that clonidine stimulates rat GH secretion, possibly by acting within the hypothalamus to stimulate GH releasing factor neurons.     

INTERACTION OF CLONIDINE AND GHRH ON GH SECRETION IN VIVO AND IN VITRO

To investigate the mechanism by which clonidine stimulates GH-secretion in vivo and in vitro, we studied its interaction with GHRH. In vivo: eight or six normal male subjects were submitted to five protocols: (1) 150 micrograms clonidine orally followed by 50 micrograms GHRH 1-44 i.v. 2 h later, (2) 50 micrograms GHRH 1-44 i.v. followed by 150 micrograms clonidine orally 2 h later, (3) 150 micrograms clonidine orally followed by GHRH i.v. 30 min later, (4) 300 micrograms clonidine orally followed by 50 micrograms GHRH i.v. 3 h later and (5) 50 micrograms GHRH i.v. followed by 300 micrograms clonidine orally 90 min later. In vitro: Rat anterior pituitary cells were coincubated with clonidine (10(-11), 10(-9), 10(-7) and 10(-5) M) and GHRH (0.005, 0.05, 10 nM) for 4 h. Results: 150 micrograms clonidine alone does not stimulate GH-secretion. Furthermore, the GH-increase was not significantly different when GHRH bolus was given before, after or together with clonidine. When 300 micrograms clonidine was given before GHRH GH-levels were significantly higher (max 28.6 +/- 8.0 mU/l) at 90 min, compared to when clonidine was given after GHRH (max 7.8 +/- 3.6 mU/l). The GHRH bolus after clonidine led to a significantly lower GH-increase (max 31.6 +/- 17.0 mU/l) compared to the GHRH-induced GH-increase (max 47.2 +/- 13.0 mU/l) before clonidine. In vitro, clonidine had no stimulatory effect on GHRH-stimulated GH secretion. These findings are compatible with clonidine leading to stimulation of GH by inducing endogenous GHRH release.     

Centrally bradykinin B2-receptor-induced hypertensive and positive chronotropic effects are mediated via activation of the sympathetic nervous system.

OBJECTIVE: The presence of bradykinin B2 receptors in the cardiovascular regulatory centres of the brain indicates that increase in mean arterial pressure (MAP) and heart rate after intracerebroventricular (i.c.v.) injections of bradykinin is mediated via stimulation of sympathetic nervous system. METHODS: Adult Wistar- Kyoto (WKY) rats were instrumented chronically with an i.c.v. cannula, and the catheters were placed into the femoral artery and vein. Increasing doses of bradykinin (1 -300 pmol) were given i.c.v. and (i) MAP and heart rate, (ii) plasma dopamine, noradrenaline and adrenaline, and (iii) plasma arginine vasopressin (AVP) levels were determined. In addition, following blockade of peripheral alpha1 -adrenoceptors with prazosin (50 and 250 microg/kg i.v.) beta1-adrenoceptors with atenolol (10 mg/kg i.v.) or V1 -receptors with TMe-AVP (Manning compound) (10 microg/kg i.c.v. and 100 microg/kg i.v.) the effects of bradykinin (100 pmol i.c.v.) on MAP and heart rate were determined. RESULTS: Bradykinin increased MAP and heart rate dose-dependently. The pressor effects of 100 pmol bradykinin i.c.v. were completely blocked by pretreatment with the specific B2 receptor antagonist Hoe 140 (3 pmol, i.c.v.). There was no change in plasma dopamine, noradrenaline, adrenaline or AVP levels after increasing doses of bradykinin. However, peripheral blockade of alpha1- and beta1-adrenoceptors reduced the bradykinin-induced increase in MAP and heart rate, whereas central and peripheral V1 receptor blockade did not alter the cardiovascular responses to i.c.v. bradykinin. CONCLUSION: Our data suggest that the hypertensive and positive chronotropic effects induced by i.c.v. bradykinin are due to stimulation of sympathoneuronal rather than sympathoadrenal pathway in vivo.     

Evidence for central and peripheral serotonergic control of corticosterone secretion in the conscious rat

Serotonin (5-HT) and 5-HT agonists act on multiple 5-HT receptor subtypes to increase corticosterone secretion. The present experiments describe the effects of a highly selective 5-HT2 receptor agonist DOI [(+-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl] on plasma corticosterone in conscious, unrestrained, male rats with indwelling arterial and venous catheters. DOI (500 micrograms/kg, i.v.) increased plasma corticosterone levels 6- to 7-fold from 15 to 60 min. Pretreatment with the central 5-HT2 antagonist LY 53857 (100 micrograms/kg, i.v.) blocked the effect of DOI on corticosterone secretion at all times. The peripheral 5-HT2 antagonist xylamidine (100 micrograms/kg, i.v.) attenuated the corticosterone response elicited 15 min after DOI but did not alter the 60-min response. In contrast, dexamethasone pretreatment (350 micrograms/kg, s.c.) attenuated the corticosterone response to DOI at 15 min, but abolished the response at 60 min. The increase in corticosterone levels elicited 5 min after the nonselective 5-HT agonist quipazine (3 mg/kg, i.v.) was also reduced by xylamidine. These data suggest that 5-HT2 receptor agonists increase corticosterone secretion initially, in part, through a direct adrenal mechanism not entirely dependent on adrenocorticotropin, and at later times via a central, dexamethasone-suppressible mechanism. This raises the possibility that endogenous 5-HT in the adrenal medulla may act as a local paracrine to participate in the regulation of corticosterone secretion from the adrenal cortex.     

Stimulation and inhibition of growth hormone secretion by interleukin-1 beta: the involvement of growth hormone-releasing hormone.

The effects of intracerebroventricularly (i.c.v.) injected interleukin-1 beta (IL-1 beta: 1, 2.5, 10, and 25 ng) were studied on plasma growth hormone (GH) and prolactin (PRL) concentrations in freely moving rats chronically implanted with i.c.v. cannulas and intracardial catheters. Significant changes in PRL secretion were not found. Small i.c.v. doses of IL-1 stimulated GH secretion 15 min postinjection (significant after 2.5 ng IL-1) whereas high doses of IL-1 suppressed plasma GH concentrations. The stimulation of GH secretion by 2.5 ng IL-1 was abolished when endogenous growth hormone-releasing hormone (GHRH) was immunoneutralized by pretreating the rats with GHRH antibodies. The results indicate that IL-1 elicits GH secretion by stimulating the release of hypothalamic GHRH. The inhibition of GH secretion after high doses of IL-1 is attributed to the previously reported corticotropin-releasing-hormone-releasing activity of IL-1.