The interaction of growth hormone releasing hormone and somatostatin in the generation of a GH pulse in man

OBJECTIVE: To study the regulation of the growth hormone (GH) response to growth hormone releasing hormone (GHRH) in the presence or absence of somatostatin pretreatment. DESIGN: Seven healthy adult male volunteers of normal height and weight and aged between 19 and 29 years underwent four separate studies (each containing three cycles in one day) in random order. The studies were separated from each other by at least a week. On day 1, three consecutive cycles (between 0800 and 2000 hours) consisted each of a saline infusion for 3 hours which was stopped prior to a bolus injection of saline and followed by 60 minutes of more intensive blood sampling. On day 2, the bolus injections were of GHRH given after saline infusion. On days 3 and 4 somatostatin infusions were administered instead of saline over the 3-hour periods followed by bolus injections of saline or GHRH respectively. In all studies, samples were collected for the measurement of serum GH concentration at 15-minute intervals from time 0 to 180 minutes and then at 5-minute intervals for a further 60 minutes, this cycle being repeated three times. MEASUREMENTS: Serum GH concentrations were analysed by serial array averaging. RESULTS: Prompt release of GH was observed in response to GHRH given against a saline background (day 2, cycle 1) (mean at 60 minutes 49.2 +/- 14.7 mU/l) but the responses observed during the second and third cycles were attenuated (mean at 60 minutes 17.2 +/- 4.0 mU/l; P = 0.025). GH release between somatostatin infusions (somatostatin withdrawal; day 3) occurred twice as often as that observed during saline infusions (62% day 3: 29% day 1). The response, although qualitatively similar to that induced by GHRH, was reduced in amplitude and the time of onset variable (5-45 minutes). On day 4, the administration of GHRH as a bolus injection combined with somatostatin withdrawal led to consistent and repeatable GH responses (mean at 60 minutes, cycle 1, 39.7 +/- 10.8 mU/l; cycles 2 and 3, 37.4 +/- 9.4 mU/l) which were similar to those observed with GHRH alone (day 2, cycle 1) (mean 39.7 +/- 10.8 mU/l) (P = NS). CONCLUSIONS: These data suggest that endogenous somatostatin secretion is important in determining the ability of the somatotroph to respond to repeated growth hormone releasing hormone stimulation and that for regular GH pulse generation a close interplay between growth hormone releasing hormone and somatostatin is required.     

Acute dexamethasone administration enhances GH responsiveness to GH releasing peptide-6 (GHRP-6) in man

OBJECTIVE: Acute administration of glucocorticoids stimulates GH secretion probably by a decrease in hypothalamic somatostatin release. GHRP-6 is a synthetic hexapeptide that increases GH secretion by a mechanism of action not yet fully known, but apparently not by inhibition of hypothalamic somatostatin release. The aim of this study was to evaluate the effect of acute dexamethasone administration on GH responsiveness to GHRP-6 in man. DESIGN: One group of subjects received iv GHRP-6 (1 microg/kg), GH-releasing hormone (GHRH; 100 microg), GHRH plus GHRP-6 or saline 3.5 h after oral acute dexamethasone administration (4 mg; at 0600 h). A second study group was treated with GHRP-6, GHRH or GHRP-6 plus GHRH after placebo ingestion, following the same protocol. PATIENTS: Sixteen normal subjects (mean age: 29 +/- 3.3 years), with normal BMI (22.4 +/- 2.0 kg/m2), were studied. Eight subjects received dexamethasone and the other eight were treated with placebo. MEASUREMENTS: Serum GH was measured by a two site monoclonal antibody immunofluorometric assay. RESULTS: In the placebo-treated subjects, mean peak GH (mU/l; mean +/- SE) and AUC (mU.min/l) values after GHRP-6 administration (peak: 43.8 +/- 9.0; AUC: 2262.0 +/- 459. 2) did not differ from those observed after GHRH injection (peak: 49. 8 +/- 12.0; AUC: 2903.4 +/- 872.6). The association of the two peptides markedly increased GH levels (peak: 172.4 +/- 34.2; AUC: 10393.0 +/- 1894.8) compared with the isolated administration of GHRP-6 or GHRH. In the subjects who received dexamethasone 3.5 h before saline injection, GH baseline values were significantly higher than those observed after 90 min of sampling (12.4 +/- 9.4 vs. 4.6 +/- 2.0). Mean GH peak and AUC values after GHRP-6 (peak: 78.8 +/- 11.0; AUC: 4114.6 +/- 588.2) and after GHRH administration (peak: 46.8 +/- 16.0; AUC: 3006.8 +/- 1010.0) did not differ significantly in the dexamethasone-treated subjects. In this study group, the administration of the two peptides together caused a significant increase in both peak (119.2 +/- 16.0) and AUC values (7377.0 +/- 937.2) compared with the response obtained after each peptide alone. When the two groups were compared, a significant increase in GH responsiveness to GHRP-6 was observed after dexamethasone administration compared with placebo. No differences in GH response to GHRH, or to the administration of the two peptides together, were seen between the two groups. CONCLUSIONS: Oral dexamethasone, at a dose of 4 mg, enhances GH releasing peptide-6-induced GH release when administered 3.5 h earlier. These results suggest that dexamethasone and GHRP-6 could act at different sites of GH releasing mechanisms. Further studies are necessary to elucidate these findings.     

Growth hormone releasing hormone 1-44 NH2 and 1-40 OH levels in normal subjects during growth hormone stimulation tests

OBJECTIVE Little is known about the relative circulating concentrations of growth hormone releasing hormone (GHRH) 1-44 NH₂ and 1-40 OH in response to dynamic GH stimulation. We therefore studied the concentrations of growth hormone-releasing hormone (GHRH) 1-44 NH₂ and 1-40 OH in the peripheral plasma of normal male subjects during GH stimulation tests. DESIGN Tests were performed at 0900 h after an overnight fast. Stimulation tests, commenced at 0 minutes, included α-adrenergic activation with adrenaline (10 μg/min from 0 to 30 minutes) following β-blockade with propranolol (1.5 mg/min from -10 to 0 minutes), α2-adrenergic activation with clonidine 150 μg i.v., insulin hypoglycaemia (0.15 U/kg soluble insulin), l -arginine infusion (30 g from 0 to 30 minutes), l -dopa (500 mg orally) and oral glucose (100 g). SUBJECTS Groups of healthy male volunteers aged 20-42 years, all within 10% of ideal body weight. MEASUREMENTS Serum GH and plasma GHRH 1-44 NH2 and 1-40 OH were measured at intervals for between 60 and 390 minutes, depending on the stimulation test. RESULTS There were no significant changes in either GHRH 1-44 or 1-40 following a-adrenergic activation with propranolol/adrenaline infusion, a2-adrenergic activation with i.v. clonidine, insulin-induced hypoglycaemia or arginine infusion despite the expected rise in GH levels. After oral glucose, GH was initially suppressed with a late rise. There were no changes in GHRH 1-44 or 1-40 levels during either phase of this response. After l -dopa GH levels peaked at 90 minutes, 24.5 ± 11.0 mU/l (mean ± SEM). At 0 minutes GHRH 1-44 and 1-40 levels were 3.25 ± 0.89 and 4.93 ± 1.28 pmol/l respectively and rose in both cases, peaking at 60 minutes at 4.23 ± 1.01 and 7.55 ± 1.80 pmol/l (P < 0.05). At no time was there any evidence of differential secretion of GHRH 1-44 or 1-40. CONCLUSIONS We have confirmed previous studies demonstrating a small rise in GHRH before the GH response to l -dopa. However, in all other situations of pharmacological stimulation of GH release we were unable to detect any significant changes in GHRH 1-44 or 1-40 levels. It seems most likely that peripheral GHRH does not reflect hypothalamic secretion. As yet there is no evidence for differential release of GHRH 1-44 and 1-40.     

Absence of desensitization by hexarelin to subsequent GH releasing hormone-mediated GH secretion in patients with anorexia nervosa

OBJECTIVEBoth the basal levels and the neuroregulation of GH secretion are perturbed in patients with anorexia nervosa. It is unknown if these alterations are due to severe undernutrition or if they reflect basic neurotransmitter alterations of the patient’s neural pathways. On the other hand, prior administration of the GH secretagogue hexarelin in normal subjects blocks the GH-releasing capability of GH releasing hormone (GHRH) administered 2 hours later. In the present work a sequential test was performed using the administration of hexarelin as first stimulus followed 120 minutes later by GHRH. The two aims of the study were: (a) to evaluate the interaction of GHRH and hexarelin, and (b) to further understand the alterations in GH neuroregulation in patients with anorexia nervosa. DESIGNThe GH stimuli used were hexarelin (1 μg/kg i.v.), a GH stimulus whose main action is hypothalamic, followed 120 minutes later by GHRH (1 μg/kg i.v.) as a pituitary stimulus. Each woman was tested once. PATIENTSThirty-two women matched for age participated in the study: six normal-weight women as controls, 14 women with anorexia nervosa, seven women with secondary amenorrhoea due to voluntary weight loss for aesthetic reasons, and five normal-weight women after 72 hours of a controlled hypocaloric diet (800 cal/day). MEASUREMENTSPlasma GH levels were measured by time-resolved fluoroimmunosasay, each value shown is the mean±SE in mU/l. RESULTSThe administration of hexarelin to the normal-weight women induced a clear-cut GH secretion (expressed as mean±SE of GH peak in mU/l of 77.5±21.8, but blocked the GH-releasing capability of GHRH administered 120 minutes later (6.6±2.8, P<0.05). In contrast, the women with anorexia nervosa showed a normal GH response after the two stimuli: hexarelin 64.8±9.2. GHRH 71.1±14.2. The absence of heterologous desensitization was specific to anorexia nervosa, because the women with amenorrhoea due to voluntary weight loss but with a normal energy intake showed a pattern similar to the controls (GH after hexarelin 60.3±9.5 and to GHRH 120 minutes later 6.2±1.0 (P<0.05)). Similarly, the women after the short-term hypocaloric diet showed a hexarelin-mediated GH secretion of 99.6±17.8, which blunted the subsequent administration of GHRH (GH mean peak of 9.9±2.9, P<0.05 vs hexarelin). CONCLUSIONSIn the normal subjects, the administration of hexarelin induced clear-cut GH secretion, but inhibited the action of GHRH when administered 120 min later, while this heterologous desensitization was not observed in the patients with anorexia nervosa. This sequentially delayed test may be of some value in the clinical setting for assessing the status of patients with anorexia nervosa.     

The GH response to low-dose bolus growth hormone-releasing hormone (GHRH(1-29)NH2) is attenuated in patients with longstanding post-irradiation GH insufficiency

OBJECTIVE: Previous studies have suggested that post-irradiation GH insufficiency results from a loss of GHRH secretion, since many patients were able to release GH following exogenous GHRH stimulation. However, supramaximal doses of GHRH were used and the response may decline with time after radiotherapy. We re-evaluated the GHRH dose-response curve in patients post cranial irradiation and in controls. DESIGN: Randomized controlled study. METHODS: Five adult male long-term survivors of childhood brain tumours (median age 21.8 years (18.4-26.7); 13.7 years (11.4-15.7) post-radiotherapy, >30Gy) and five matched controls were studied. An intravenous bolus of GHRH(1-29)NH(2) was administered in doses at the lower (0.05 microg/kg) and upper (0.15 microg/kg) range of the dose-response curves for young males, as well as the standard supramaximal dose (1. 0 microg/kg). GH was measured before stimulation, every 2min for the first hour and every 5min for the second hour. All studies were conducted in a random fashion. RESULTS: Significantly lower peak and area under the curve (AUC) GH concentrations occurred in the irradiated group using 0.15 microg/kg (median peak Irradiated, 4. 5mU/l vs median Controls, 37.4mU/l; P<0.01) and 1.0 microg/kg (median peak Irradiated, 4.8mU/l vs median Controls, 15.2mU/l; P<0. 05) GHRH(1-29)NH(2). In irradiated subjects there was an incremental rise in GH output with increasing doses of GHRH(1-29)NH(2 )(median AUC: 122mU/l.min vs 179mU/l.min vs 268mU/l.min; P=0.007) reflecting altered pituitary sensitivity and reduced responsiveness. CONCLUSION: The GH response to bolus GHRH(1-29)NH(2) is attenuated in adult long-term survivors of childhood brain tumours. This may reflect direct pituitary damage and/or the loss of the tropic effects of chronic GHRH deficiency.     

Inhibition of growth hormone release after the combined administration of CHRH and GHRP-6 in patients with Cushing's syndrome

OBJECTIVE In patients with Cushing's syndrome there is a blunted OH response to all types of stimuli. Although Inferential data point towards a direct perturbation in the pituitary exerted by glucocorticoids, the bask mechanism is unknown. His-d -TRP-ALA-TRP-d -Phe-Lys-NH₂ (GHRP-6) is a synthetic hexapeptlde which releases GH by a direct pituitary effect through receptors other than GHRH receptors. Furthermore, the combined administration of GHRH and GHRP-6 is able to induce a large OH discharge even in some pathological states such as obesity, associated with GH blockade. To gain further insight into the disrupted mechanisms of GH secretion, Cushing's syndrome patients were challenged with either GHRH, GHRP-6 or GHRH together with GHRP-6. A group of normal subjects was included for control purposes. DESIGN Three different tests were undertaken: (a) GHRH 100 μg I.v.; (b) GHRP-6 100 μg I.v. and (c) GHRH plus GHRP-6 100 μg I.v. of each; administered to each subject on different days, at least 4 days apart. PATIENTS Ten patients (8 women, 2 men) with untreated Cushing's syndrome, 9 Cushing's disease and 1 adrenal adenoma. Five healthy volunteers (3 women, 2 men) of similar ages served as a control group. MEASUREMENTS Plasma OH levels were measured by immunoradiometric assay. RESULTS The areas under the curve (AUC) of OH secretion (mean ± SEM In μ/1/120 mi) in the control subjects after each test were: GHRH, 1420 ± 330; GHRP-6, 2278 ± 290 and GHRH plus GHRP-6,7332 ± 592 (P < 0·05 vs each compound alone). The AUCs for Cushing's syndrome patients were: GHRH, 248 ± 165; GHRP-6 530 ±170 and for GHRH plus GHRP-6, 870 ± 258 (P < 0·05 vs GHRH alone). After the combined stimulus only one out of the ten patients with hypercortisolism showed a GH peak over 20 μ/l, while ail the controls had a peak over 04mU/l. CONCLUSIONS GHRP-6 induced OH secretion as well as the OH discharge elicited by GHRH and GHRP-6 are considerably reduced in Cushing's syndrome patients. This suggests that the main impairment of GH secretion in that pathological state resides at pituitary level.     

Complementary secretagogue pairs unmask prominent gender-related contrasts in mechanisms of growth hormone pulse renewal in young adults

The present study examines the thesis that pulsatile GH secretion is controlled simultaneously by three principal signals; viz., GHRH, GH-releasing peptide (GHRP, ghrelin), and somatostatin (SS). According to this ensemble notion, no single regulatory peptide acts alone or can be interpreted in isolation. Therefore, to investigate gender-specific control of pulsatile GH secretion, we designed dual-effector stimulation paradigms in eight young men and six women as follows: 1) L-arginine/GHRH (to clamp low SS and high GHRH input); 2) L-arginine/GHRP-2 (to clamp low SS and high GHRP drive); 3) GHRH/GHRP-2 (to clamp high GHRH and high GHRP feedforward); vs. 4) saline (unclamped). Statistical comparisons revealed that: 1) fasting pulsatile GH secretion was 7.6-fold higher in women than men (P < 0.001); 2) L-arginine/GHRH and L-arginine/GHRP-2 evoked, respectively, 4.6- and 2.2-fold greater burst-like GH release in women than men (P < 0.001 and P = 0.015); and 3) GHRH/GHRP-2 elicited comparable GH secretion by gender. In the combined cohorts, estradiol concentrations positively predicted responses to L-arginine/GHRP-2 (r2= 0.49, P = 0.005), whereas testosterone negatively predicted those to L-arginine/GHRH (r2= 0.56, P = 0.002). Based upon a simplified biomathematical model of three-peptide control, the current outcomes suggest that women maintain greater GHRH potency, GHRP efficacy, and opposing SS outflow than men. This inference upholds recent clinical precedence and yields valid predictions of sex differences in self-renewable GH pulsatility.     

Growth hormone/insulin-like growth factor-1 response to acute and chronic growth hormone-releasing peptide-2, growth hormone-releasing hormone 1-44NH2 and in combination in older men and women with decreased growth hormone secretion

To better appreciate the interactions of GHRP-2 and GHRH 1-44NH₂ on the release of GH in normal adult men and women with decreased GH secretion and low serum IGF-1 levels, a series of acute and chronic studies have been performed (n=5 men, 5 women). The acute iv bolus GH responses of these subjects to the two peptides alone and together suggest that the decreased GH secretion may be primarily due to a deficiency of the natural endogenous GHRP, ghrelin, rather than a decreased secretion of endogenous GHRH or excess secretion of SRIF. To determine whether the low GH response to GHRH was due to a limited capacity of pituitary to release GH, higher dosages of GHRP-2 alone were administered. At a dose of 1 μg/kg GHRP-2 the GH response was essentially the same as that elicited by 1 μg/kg GHRH+0.1μg/kg GHRP-2 while the GH response to 10 μg/kg GHRP-2 sc was about twice as high in both men and women. Although these subjects have a limited pituitary capacity to release GH, which is also an indication of decreased GH secretion in the presence of low serum IGF-1 levels, this alone would not explain the low GH response to GHRH. Furthermore, the finding that a low dose of 0.1 μg/kg GHRP-2 augments the GH response to 1 μg/kg GHRH is strongly against an excess secretion of SRIF. Twenty-four hour profiles of GH secretion during placebo, GHRP-2, and various doses of GHRH alone and together with GHRP-2 were studied. In addition, 1 μg/kg/h GHRP-2 was infused continuously sc to these subjects for 30 d. The normal pulsatile secretion of GH as well as the serum IGF-1 level was increased after 24 h and remained elevated for 30 d. With a deficiency of endogenous GHRH, the GH response of GHRP-2 would be little to none, while in subjects with a deficiency of the natural GHRP, the GH response to GHRH would be more attenuated. Thus, in chronic deficiency the GH response would be expected to depend on the degree of the capacity of the pituitary to release GH as well as the type(s) of hormonal deficiency.     

Growth hormone response to GHRH + GHRP-6 in type 2 diabetes during euglycemic and hyperglycemic clamp

The aim of this study was to investigate the effect of two different glucose levels on GH response to the combined administration of GHRH+GHRP-6 in patients with type 2 diabetes. GH response to i.v. bolus of GHRH+GHRP-6 (100 mcg, each) was measured in 12 male patients with type 2 diabetes (mean age: 53.9+/-1.59 years; BMI: 25.58+/-0.39 kg/m(2); mean HbA(1c): 8.7+/-0.42%), during a euglycemic (mean glucose: 4.92+/-0.08 mmol) hyperinsulinemic clamp (insulin infusion rate of 100 mU/kg/h) and a hyperglycemic clamp (mean glucose: 12.19+/-0.11 mmol/l). There was no difference in basal GH levels between the hyperglycemic and euglycemic clamps (2.9+/-0.99 mU/l versus 1.48+/-0.44 mU/l; P>0.05). Peak GH response to GHRH+GHRP-6 during the hyperglycemic clamp was lower than in the englycemic clamp (112.45+/-14.45 mU/l versus 151.06+/-16.87 mU/l; P<0.05). Area under the GH curve was lower in the hyperglycemic than in the euglycemic clamp (6974.49+/-1001.95 mU/l/min versus 9560.75+/-1140.65 mU/l/min; P<0.05). It is concluded that hyperglycemia significantly reduces GH response to combined administration of GHRH+GHRP-6 in normal weight patients with type 2 diabetes. It is suggested that ambient glucose levels should be taken into account during interpretation of GH response to combined administration of GHRH+GHRP-6 in patients with type 2 diabetes.     

Growth hormone (GH)-releasing peptide stimulates GH release in normal men and acts synergistically with GH-releasing hormone

The acute GH release stimulated by the synthetic hexapeptide, His-DTrp-Ala-Trp-DPhe-Lys-NH2 [GH releasing peptide (GHRP)], was determined in 18 normal men and compared with the effects of GH-releasing hormone, GHRH-(1-44)-NH2. Specificity of effect was assessed by measurement of serum PRL, LH, TSH, and cortisol. GHRP was administered at doses of 0.1, 0.3, and 1.0 microgram/kg by iv bolus. GHRH at a dose of 1.0 microgram/kg was administered alone and together with various does of GHRP. No adverse clinical effects of laboratory abnormalities were observed in response to GHRP. A side-effect of mild facial flushing of 1- to 3-min duration occurred in 16 of the 18 subjects who received GHRH-(1-44)-NH2. Mean (+/- SEM) peak serum GH levels after injection of placebo and 0.1, 0.3, and 1.0 microgram/kg GHRP were 1.2 +/- 0.3, 7.6 +/- 2.5, 16.5 +/- 4.1, and 68.7 +/- 15.5 micrograms/L, respectively. The submaximal dosages of 0.1 and 0.3 microgram/kg GHRP plus 1 microgram/kg GHRH stimulated GH release synergistically. Serum PRL and cortisol levels rose about 2-fold above basal levels only at the 1 microgram/kg dose of GHRP, and there were no changes in serum LH and TSH over the first hour after administration of the peptide(s). GHRP is a potent secretagogue of GH in normal men. Since GHRP and GHRH together stimulate GH release synergistically, these results suggest that GHRP and GHRH act independently. This supports our hypothesis that the GH-releasing activity of GHRP reflects a new physiological system in need of further characterization in animals and man.     

A single growth hormone (GH) determination is sufficient for the diagnosis of GH-deficiency in adult patients using the growth hormone releasing hormone plus growth hormone releasing peptide-6 test

BACKGROUND: The diagnosis of GH deficiency in adults is based on the provocative testing of GH secretion. When testing a patient with suspected GH deficiency, clinicians assess the whole secretory curve and select the GH peak as an index of secretory capability. This procedure is time consuming and the determination of GH in several samples is necessary. The combined administration of growth hormone releasing hormone (GHRH) plus growth hormone releasing peptide-6 (GHRP-6) is an effective test of GH secretion, and it has been unambiguously demonstrated that the elicited GH peak is capable of segregating normal GH secretion subjects from GH deficient patients on an individual basis. The GHRH + GHRP-6 test biochemically classifies patients into three groups; those with a stimulated GH peak >/= 20 micro g/l are considered normal and those with peaks at 相似文献   

The sequential administration of growth hormone-releasing hormone followed 120 minutes later by hexarelin, as an effective test to assess the pituitary GH reserve in man

OBJECTIVE GH deficiency, either in children or in adults, is a clinically relevant problem. The diagnosis is based on dynamic tests of GH secretion, which are clear cut on a group basis but highly problematic for individual diagnosis. The controversy surrounding the diagnosis of GH deficiency reflects the absence of a gold standard dynamic test. The synthetic hexapeptide hexarelin and GHRH stimulate GH secretion using different mechanisms. A sequential test has been devised using the administration of GHRH as first stimulus followed 120 minutes later by hexarelin. The two aims of the study were (a) to evaluate the interaction of GHRH and hexarelin, and (b) to devise a sequential test of GH reserve. DESIGN The GH stimuli used were GHRH (1 μg/kg i.v.) as a pituitary stimulus, and hexarelin (1 μg/kg i.v.) as a GH stimulus whose main action is hypothalamic. Each subject was tested twice in order to serve as his own control. Three different studies, each with two duplicate tests, were performed on separate groups of individuals: (a) GHRH followed 120 minutes later by hexarelin and on the second day hexarelin followed 120 minutes later by GHRH; (b) GHRH followed 120 minutes later by GHRH and on the other day hexarelin followed 120 minutes later by hexarelin; (c) GH 0.5 IU i.v. followed 120 minutes later by GHRH and on the other day, the same dose of GH followed 120 minutes later by hexarelin. PATIENTS Eighteen normal volunteers (12 women, 6 men) after giving informed consent. MEASUREMENTS Plasma GH levels were measured by time-resolved fluoroimmunoassay; each value shown is the mean ± SEM of n = 6. RESULTS GHRH followed 120 minutes later by hexarelin induced two episodes of GH secretion (expressed as mean GH peak, mU/l). The GHRH-mediated GH release showed a mean GH peak of 38.2 ± 13.6 mU/l and after hexarelin 120 minutes later of 56.7 ± 18.0 mU/l. The contrary sequence blocked the second stimulus, i.e. the hexarelin-stimulated GH mean peak was 54.7 ± 18.4, and the GH release 120 minutes later after GHRH was 4.8 ± 1.9 (P < 0.05 vs GHRH used as first stimulus). In the two sequential tests using the same stimulus, the second GH peak was reduced. In fact, GHRH induced a GH mean peak of 63.8 ± 21.1 mU/l as first stimulus, greater (P < 0.05) than when GHRH was administered again 120 minutes later (22.0 ± 5.9 mU/l). Similar results were obtained with hexarelin, with a first mean peak of 70.6 ± 10.3 mU/l, and a second one 120 minutes later of 13.4 ± 4.6 mU/l (P < 0.05). The blockade of the second stimulus was not due to the feed-back action of the GH released by the first stimulus. In fact, the i.v. administration of exogenous GH induced a mean GH peak of 168.0 ± 89.7 and reduced the action of GHRH administered 120 minutes later (26.1 ± 8.1). The previous administration of GH (mean peak 115.5 ± 42.0) did not alter the action of hexarelin injected 120 minutes later, showing a mean peak of 71.9 ± 11.2. The large variability in the stimulatory action of GHRH contrasted vividly with the reproducibility of hexarelin. Furthermore, individually analysed, only one of the 12 subjects tested first with hexarelin, compared to 4 out of 12 tested with GHRH as first stimulus, presented a blunted response (<13 mU/l). After the sequential stimulus there were no false negatives. CONCLUSION The sequential administration of GHRH in normal subjects and of hexarelin 120 minutes later provides separate information regarding pituitary GH reserve, of both secretagogues without mutual interference. There were not false negative results to the combined test. This sequentially delayed test may be of some value in the clinical setting for assessing pituitary GH reserve.     

Growth hormone secretion after the administration of GHRP-6 or GHRH combined with GHRP-6 does not decline in late adulthood

OBJECTIVE Growth hormone (GH) secretion in middle and late adulthood declines with age. However, the precise mechanisms causing this impairment in OH release are unknown. HIs-D-Trp-Ala-Trp-D-Phe-Lys-NH₂(GHRP-6) Is a synthetic compound that releases OH in a dose related and specific manner in several species, including man. In order to gain a further Insight Into disrupted GH secretion in late adulthood, we evaluated GH responses to GHRP-6 or GHRH, administered either alone or in combination, in healthy young and late adulthood groups of subjects. DESIGN All subjects underwent three different tests carried out in random order and separated by at least one week. Tests were performed at 0900 h after an overnight fast. GHRH (100 μg), GHRP-6 (90 μg) either alone or In combination were administered as an I.v. bolus. SUBJECTS Groups of healthy young (mean ± SEM 22 ± 1·1 years, n= 9) and older adult subjects (59·5 ± 1·7 years, n= 9) were studied. MEASUREMENTS Serum GH levels were measured by radioimmunoassay. RESULTS In the group of young adult subjects the combined administration of GHRH and GHRP-6 elicited a greater GH increase than GHRH alone (F= 21·9, P < 0·001) or GHRP-6 alone (F= 6·2, P= 0·01). Similarly, the response to the combined stimuli was also greater than with GHRH alone (F= 21·8, P < 0·001) or GHRP-6 alone (F= 23·9, P < 0·001) In the late adulthood group of subjects. OH responses to GHRH were greater in younger than in older subjects (F= 3·45, P= 0·03). In contrast, OH responses to either GHRP-6 (F= 0·71, P= NS) or combined GHRH plus GHRP-6 administration (F= 0·68, P= NS) were not significantly different between the two groups. CONCLUSIONS These data show that OH responses to GHRP-6 are much greater than to GHRH In late adulthood. The marked Increase of plasma GH levels observed after administration of GHRP-6 alone or in combination with GHRH Indicates that Impaired GH secretion in late adulthood is a functional and potentially reversible state.     

The impact of cranial irradiation on GH responsiveness to GHRH plus GH-releasing peptide-6

Patients treated with cranial radiation are at risk of GH deficiency (GHD). We evaluated somatotroph responsiveness to maximal provocative tests exploring the GH releasable pool in relation to the impact of radiation damage to the hypothalamic-pituitary unit. The GH-releasing effect of GHRH plus GH secretagogue [GH-releasing peptide (GHRP)-6] (GHRH+GHRP-6) was studied in 22 adult patients (age, 23.2 +/- 1.4 yr; 8 female and 14 male; mean body mass index, 22.6 +/- 0.7 kg/m(2)) who received cranial radiation for primary brain tumor distant from hypothalamic-pituitary region 7.6 +/- 0.7 yr before GH testing. Two stimulatory tests for GH secretion were employed: insulin tolerance test (ITT, 0.15 IU/kg regular insulin i.v. bolus); and GHRH+GHRP-6 test: GHRH (Geref Serono, Madrid, Spain; l microg/kg) plus GHRP-6 (CLINALFA, Laufelingen, Switzerland; 1 microg/kg) as i.v. bolus. Serum GH was measured (Delphia; Perkin Elmer, Wallac, Turku, Finland) at -30, -15, 0, 15, 30, 45, 60, 90, and 120 min. Anterior pituitary function was normal in all except in 1 female with hyperprolactinemia. Twelve out of 22 irradiated patients were GH-deficient (GHD) with both tests. Eleven out of 22 (50%) irradiated patients were severely GHD, according to the ITT (GH < 3 microg/liter; mean GH peak, 1.5 +/- 0.5 microg/liter). In 9 irradiated patients, in whom ITT was performed as well, mean peak GH after the GHRH+GHRP-6 test was 6.2 +/- 0.8 microg/liter, which is considered as severe GHD, according to our own cut-off for the test (peak GH < 10 microg/liter). GH responses to both tests were highly concordant, but the differential in the GH peak concentrations between GHD and non-GHD irradiated patients was significantly larger for the GHRH+GHRP-6 test than that for the ITT. The 2 discordant responses, i.e. poor response to the ITT and good response to the GHRH+GHRP-6 test, were found in 1 hyperprolactinemic female patient and in 1 other female. One irradiated patient was diagnosed as GHD only with the combined test, because ITT was contraindicated because of epilepsy. PRL and cortisol responses to ITT were normal in all irradiated patients and did not depend on the GH status. IGF-I levels were not informative or discriminative between the GHD and non-GHD irradiated adult patients. In conclusion, the use of GH secretagogues plus GHRH is an easy, reliable and accurate way of assessing GH secretion in cranially irradiated patients. Impairment of the GH releasable pool in the irradiated patients, with a maximal provocative test, reflects alterations in the hypothalamic-pituitary unit caused by radiotherapy.     

Acute growth hormone (GH) response to GH-releasing hexapeptide in humans is independent of endogenous GH-releasing hormone.

The synthetic GH-releasing hexapeptide (GHRP: His-DTrp-Ala-Trp-DPhe-Lys-NH2) releases GH in man by an undetermined mechanism. To investigate whether acute GH response to GHRP is mediated by endogenous GHRH, we examined the effect of GHRP on GH release during pituitary desensitization to GHRH induced by short-term GHRH infusion. In five healthy men on six occasions, we infused saline (sal) or 1 microgram/kg.h GHRH-44 for 6 h. After 4 h, a bolus of sal, GHRH-44 1 microgram/kg body weight, or GHRP 1 microgram/kg body weight was given iv. GH concentration, measured by RIA, was analyzed by mean area under the curve (AUC) of GH released over the 2 h immediately after bolus injection. Infusion of GHRH had a biphasic effect on GH release; plasma GH increased to 12.7 +/- 3.3 micrograms/L within the first hour, with subsequent decrease to 2.9 +/- 0.3 micrograms/L during the last 2 h of infusion. GH AUC (hours 4-6 of infusion) microgram/L.2 h [table: see text] GH response to bolus GHRH was abolished by GHRH infusion, whereas GH response to GHRP persisted under the same conditions. Thus, we conclude that acute GH response to GHRP in humans is not mediated by endogenous GHRH.     

Physical activity or food intake prior to testing did not affect the reproducibility of GH secretion elicited by GH releasing hormone plus GH-releasing hexapeptide in normal adult subjects

OBJECTIVE: Growth hormone deficiency (GHD) in adults is a defined syndrome of which the adverse effects on different areas of body function are reversed under replacement therapy with GH. The diagnosis of GHD is controversial in adults, relying on the GH secretion elicited by the so-called provocative tests of GH reserve. Most of the tests in use, including the widely employed insulin tolerance test, have been shown to be blunted after daily activities, such as mild exercise, heat or food intake, which makes stringent testing conditions mandatory in order to assure reproducibility. The combined administration of GH releasing hormone (GHRH) and GH-releasing hexapeptide (GHRP-6) is a very effective test for the diagnosis of GH deficiency in adults. In the present study, the perturbatory action of mild physical activity and food intake on the reproducibility of this combined test was assessed. METHODS: Seventeen healthy volunteers of both sexes were tested twice on separate occasions with the sequential administration of GHRH (90 microg i.v.) plus GHRP-6 (90 microg i.v.) as bolus. Eleven subjects underwent the first combined test in the morning under basal conditions and the second test was performed in the afternoon of the same day after a morning of habitual working activity and after a standard lunch. Another group of six subjects underwent similar double testing in the morning and in the afternoon after morning activity followed by lunch; however, both tests were separated by a period of 6 months. MEASUREMENTS: GH levels were analysed by time-resolved fluoroimmunoassay, with sampling every 15 minutes. RESULTS: The reproducibility of the GHRH + GHRP-6 test performed on the same day was high, with the mean GH peak being 65.4 +/- 8.0 microg/l in the basal state and 51.2 +/- 9.6 microg/l after lunch, while the area under the curve (AUC) was 2287 +/- 288 microg/l per 45 minutes in the morning and 1759 +/- 350 after lunch. When a period of 6 months had elapsed between the first and the second test, the reproducibility was well maintained with a mean GH peak of 42.7 +/- 6.2 microg/l in the basal state and 34.3 +/- 3.6 microg/l in the after lunch test, with the AUC 1463 +/- 209 and 1164 +/- 106 microg/l per 45 minutes in the morning and after lunch, respectively. When analysed individually, physical activity, lunch or time elapsed between the two tests did not significantly change the GH peak in the subjects tested. CONCLUSIONS: The GHRH + GHRP-6 test of GH reserve is a highly reproducible test in adult subjects over time, and is not perturbed by common daily activities, such as mild physical exercise or food intake. Considering that stringent testing conditions are not required and the whole test may be performed in 30 minutes, it may be useful in the clinical setting for the diagnosis of GH deficiency in adults.     

The effect of repeated administration of hexarelin, a growth hormone releasing peptide, and growth hormone releasing hormone on growth hormone responsivity

OBJECTIVE Hexarelin is a synthetic six-amino-acid compound capable of releasing GH in animals and in man. Its mechanism of action is not understood and little is known about the GH response after repeated administration. The aim of this study was to determine the GH response to the administration of two intravenous boluses of hexarelin, growth hormone releasing hormone (GHRH) or hexarelin with GHRH. DESIGN Single boluses of hexarelin (1 μg/kg), GHRH-(1–29)-NH₂ (1 μg/kg) or hexarelin with GHRH-(1–29)-NH₂ were administered intravenously. Each study was performed on two further occasions, with a second bolus being administered 60 or 120 minutes after the first. A control study was performed giving saline intravenously. Studies were performed in a random order. SUBJECTS Six healthy adult males (25.4–34.1 years) were studied. MEASUREMENTS Serum GH was measured by radioimmunoassay. GH secretion rates were derived from the measured serum GH concentrations using the technique of deconvolution analysis. RESULTS The peak GH secretion rate following the first intravenous bolus of hexarelin was greater than that following the first bolus of GHRH-(1–29)-NH₂ (P < 0.001), and was greatest following the administration of hexarelin with GHRH-(1–29)-NH₂ (P < 0.001). The coadministration of the two secretagogues resulted in peak GH secretion rates significantly greater than the arithmetic sum of those following their isolated administration (P = 0.001), demonstrating synergism. Compared to saline, the administration of a second bolus of hexarelin, GHRH-(1–29)-NH₂ or both resulted in significant further GH secretion (P = 0.02, P = 0.002, P = 0.03, respectively). The administration of a second bolus of hexarelin or hexarelin with GHRH-(1–29)-NH₂ 120 minutes after the first bolus resulted in lower peak GH secretion rates (P = 0.03). The reductions in peak GH secretion rates following the 60-minute boluses were not statistically significant. The peak GH secretion rates following the first GHRH-(1–29)-NH₂ boluses were similar to those following the 60 and 120-minute GHRH-(1–29)-NH₂ boluses (P = NS). Irrespective of the interval between the boluses of hexarelin with GHRH-(1–29)-NH₂, the peak GH secretion rates following the second boluses were not significantly different from the arithmetic sum of those following the administration of the second boluses of hexarelin or GHRH-(1–29)-NH₂, indicating loss of synergism on repeated administration. CONCLUSIONS This study shows that hexarelin is a potent GH secretagogue active after two successive doses; the magnitude of the GH response to the second dose was influenced by the dosing interval. Hexarelin and GHRH-(1–29)-NH₂ are synergistic, a property which is lost after repeated administration. These findings may help our understanding of GHRPs and may have implications for the potential use of hexarelin and other GHRPs as therapeutic agents.     

Pituitary responsiveness to GH-releasing hormone, GH-releasing peptide-2 and thyrotrophin-releasing hormone in critical illness

OBJECTIVE Protein hypercatabolism and preservation of fat depots are hallmarks of critical illness, which is associated with blunted pulsatile GH secretion and low circulating IGF-I, TSH, T4 and T3. Repetitive TRH administration is known to reactivate the pituitary-thyroid axis and to evoke paradoxical GH release in critical illness. We further explored the hypothalamic-pituitary function in critical illness by examining the effects of GH-releasing hormone (GHRH) and/or GH-releasing peptide-2 (GHRP-2) and TRH administration. PATIENTS AND DESIGN Critically ill adults (n=40; mean age 55 years) received two i.v. boluses with a 6-hour interval (0900 and 1500 h) within a cross-over design. Patients were randomized to receive consecutively placebo and GHRP-2 (n=10), GHRH and GHRP-2 (n=10), GHRP-2 and GHRH+GHRP-2 (n=10), GHRH+GHRP-2 and GHRH+GHRP-2+TRH (n=10). The GHRH and GHRP-2 doses were 1μg/kg and the TRH dose was 200μg. Blood samples were obtained before and 20, 40, 60 and 120 minutes after each injection. MEASUREMENTS Serum concentrations of GH, T4, T3, rT3, thyroid hormone binding globulin (TBG), IGF-I, insulin and cortisol were measured by RIA; PRL and TSH concentrations were determined by IRMA. RESULTS Critically ill patients presented a striking GH response to GHRP-2 (mean±SEM peak GH 51±9 μg/l in older patients and 102±2μg/l in younger patients; P=0.005 vs placebo). The mean GH response to GHRP-2 was more than fourfold higher than to GHRH (P=0.007). In turn, the mean GH response to GHRH+GHRP-2 was 2.5-fold higher than to GHRP-2 alone (P=0.01), indicating synergism. Adding TRH to the GHRH+GHRP-2 combination slightly blunted this mean response by 18% (P=0.01). GHRP-2 had no effect on serum TSH concentrations whereas both GHRH and GHRH+GHRP-2 evoked an increase in peak TSH levels of 53 and 32% respectively. The addition of TRH further increased this TSH response < ninefold (P=0.005), elicited a 60% rise in serum T3 (P=0.01) and an 18% increase in T4 (P=0.005) levels, without altering rT3 or TBG levels. GHRH and/or GHRP-2 induced a small increase in serum PRL levels. The addition of TRH magnified the PRL response 2.4-fold (P=0.007). GHRP-2 increased basal serum cortisol levels (531±29nmol/l) by 35% (P=0.02); GHRH provoked no additional response, but adding TRH further increased the cortisol response by 20% (P=0.05). CONCLUSIONS The specific character of hypothalamic-pituitary function in critical illness is herewith extended to the responsiveness to GHRH and/or GHRP-2 and TRH. The observation of striking bursts of GH secretion elicited by GHRP-2 and particularly by GHRH+GHRP-2 in patients with low spontaneous GH peaks opens the possibility of therapeutic perspectives for GH secretagogues in critical care medicine.     

Cholinergic control of growth hormone (GH) responses to GH-releasing hormone in insulin dependent diabetics: evidence for attenuated hypothalamic somatostatinergic tone and decreased GH autofeedback

OBJECTIVE We have investigated the effects of cholinergic modulation with pirenzepine and pyridostigmine and of GH pretreatment on the subsequent GH response to a maximal stimulatory dose of GH-releasing hormone (GHRH) in patients with insulin dependent diabetes mellitus (IDDM). We have also investigated the relationship between the differences in metabolic control and other parameters of disease state with the differences in GH responses in IDDM. PATIENTS Thirteen male subjects with IDDM and no clinical evidence of complications were selected based on HbA, levels to provide a wide range of metabolic control. Seven normal subjects were also studied. DESIGN Twelve of the subjects with IDDM and six normal subjects received pirenzepine 200 mg and pyriostigmine 120 mg pretreatment 60 minutes and GH pretreatment 3 hours before an i.v. injection of GHRH (1–44) (80 μg) in random order. All subjects underwent a control study with GHRH alone. MEASUREMENTS Serum GH and plasma glucose were measured at regular intervals throughout the study. Fasting plasma glucose and HbA, were measured before each study to provide measures of metabolic control. RESULTS Subjects with IDDM demonstrated exaggerated GH responses to GHRH compared to normals. Pirenzepine significantly reduced GH responses in both normal and diabetic subjects. However, the GH response to GHRH after pirenzepine was higher in subjects with IDDM (mean GH: IDDM vs normals; 8.1 ± 1.3 vs 2.9 ± 0.7 mU/l, P < 0.05). Pyridostigmine 120 mg significantly augmented the GH response to GHRH in normal subjects. In diabetic subjects, pyridostigmine failed to increase GH response to GHRH compared to GHRH alone (mean GH: pyridostigmine vs control: 75.7 ± 12.6 vs 38.9 ± 5.4 mU/l, P= NS). GH responses to GHRH after pyridostigmine pretreatment in both normal and diabetic subjects did not differ and the GH response to GHRH after pyridostigmine in normal subjects did not differ from the GH response to GHRH alone in diabetic subjects. In normal subjects, GH pretreatment significantly reduced subsequent GH responsiveness to GHRH (Δpeak GH 26.4 ± 5.2 vs 7.7 ± 5.4 mU/I, P< 0.04). In contrast, GH pretreatment did not cause any significant reduction in GH responsiveness to GHRH In diabetics (Δpeak GH 53.6 ± 9.7 vs 33.4 ± 11 mU/I, P= NS). No significant correlation was demonstrated between measures of diabetic control and the responses to GHRH alone or after cholinergic modulation and GH pretreatment. CONCLUSION These data suggest that ambient hypothalamic cholinergic tone in diabetes is high, and of similar degree to the enhanced cholinergic tone in normal subjects pretreated with pyridostigmine. We suggest that in diabetic subjects, the reduced responsiveness to autofeedback may be secondary to the enhanced cholinergic tone demonstrated in these patients. The mechanisms linking the uncontrolled diabetic state to this abnormal neuroregulation of GH remains unknown at present.