Human Pancreatic Growth Hormone Releasing Research Articles

Growth hormone releasing factor infusion does not sustain elevated GH-levels in normal subjects.

To evaluate the dynamics of GH-secretion after infusion of growth hormone releasing factor, human pancreatic growth hormone releasing factor (hpGRF1-44) was infused over 2 and 5 h at a dosage of 100 micrograms hpGRF1-44/h into 11 healthy subjects. The infusion was started and terminated with a 50 micrograms hpGRF1-44 bolus injection. In 5 subjects 200 micrograms TRH was given 4 h after starting the infusion. In addition, 4 healthy subjects received 50 micrograms hpGRF1-44 bolus injection every 2 h. GRF, somatostatin, GH, Prl, and TSH were measured by radioimmunoassay. The initial 50 micrograms GRF bolus increased GH-levels in all 11 subjects with a maximum at 30 min (24.1 +/- 5.1 ng/ml +/- SE). However, though hpGRF1-44 was continuously infused and GFR-levels remained elevated, GH decreased to a minimum 270 min after start of infusion (2.6 +/- 0.6 ng/ml). The GH-response to the second bolus at the end of the infusion was lower compared to the first response (14.6 +/- 3.4 ng/ml after 2 h and 7.6 +/- 2.5 ng/ml after 5 h). TRH did not lead to a GH-increase during hpGRF1-44 infusion though Prl and TSH rose normally. The intermittent bolus injection of 50 micrograms hpGRF1-44 led to continuously decreasing GH-responses to the same GRF-dosage (I. bolus: 16.5 +/- 1.6 ng/ml; II. bolus: 4.2 +/- 0.8 ng/ml; III. bolus: 3.4 +/- 0.5 ng/ml). No change in somatostatin levels was observed. These findings show that GRF infusion or bolus injection in short intervals does not sustain elevated GH-levels.(ABSTRACT TRUNCATED AT 250 WORDS)

The response of normal subjects and patients with idiopathic growth hormone deficiency to continuous infusion of human pancreatic growth hormone releasing factor 1-44.

The GH response to a prolonged continuous infusion of human pancreatic growth hormone releasing factor (hpGRF 1-44), 0.3 microgram/kg/h was evaluated in seven patients with severe idiopathic GH deficiency, one patient with partial deficiency and in seven normal controls. Normal controls had a multiphasic response with a first peak between 15 and 90 min of 13.6 to 88.5, mean 45.6 mU/l and a second peak between 105 and 195 min of 8.8 to 73.1, mean 33.8 mU/l. However, the magnitude and pattern of response were highly variable. Six of the seven patients with severe GH deficiency had a response with a maximum level between 30 and 90 min of 3.3 to 7.5, mean 4.8 mU/l. Mean levels remained greater than 2.0 mU/l throughout the infusion but further peaks were absent or minimal. The patient with partial GH deficiency responded in a normal manner. hpGRF 1-44 by continuous infusion thus induced GH release in seven of eight patients with idiopathic GH deficiency though the response was impaired and the pattern of secretion abnormal in severely deficient patients.

Immunohistochemical identification of growth hormone-releasing factor-like material in the nervous system of an insect, Aeshna cyanea (Odonata)

Cells immunoreactive to antibodies raised against human pancreatic growth hormone releasing factor 1–44−NH 2 (hp GRF) were detected in the brain and the suboesophageal ganglion of an insect. The presence of immunoreactive deposits within the insect neurohaemal organ, the corpora cardiaca and within the nervi corporis cardiaci which, at least, transfers part of the neurosecretory products of the brain to the corpora cardiaca, may indicate the participation of GRF-like substance in some neurohormonal function (s) in addition to having probably a neurotransmitter role within the nervous system.

Characterization of the relaxant effects of vasoactive intestinal peptide (VIP) and PHI on isolated brain arteries

We have studied the vasorelaxant properties of vasoactive intestinal peptide (VIP) using helical strips of bovine, porcine and human brain arteries in vitro. The resting tension of the arterial strips was increased during experiments by prostaglandin F 2α or KCl so as to increase the magnitude of the relaxant response to VIP. Arteries supplying different regions of the bovine brain responded potently to VIP with ED 50 values of 1.8 nM, 2.3 nM, 6.8 nM and 9.0 nM for the middle, anterior and posterior cerebral arteries and the basilar artery, respectively. The porcine basilar artery and branches of the human middle cerebral artery responded to VIP with ED 50 values of 4.2 nM and 1.6 nM, respectively. The homologous neuropeptide, PHI, relaxed the bovine middle cerebral and porcine basilar arteries less potently than did VIP, with ED 50 values for PHI being 11 nM and 43 nM, respectively. However, PHI elicited in the two arteries a maximal vasodilatory response of similar magnitude as did VIP. The other homologous peptides, human pancreatic growth hormone releasing factor 1–40 [hpGRF 1–40], secretin, and glucagon, and the VIP fragments, VIP 1–12 and VIP 10–28, were completely inactive. In constrast, VIP, which had been oxidized to VIP-(Met 17 sulfoxide) or VIP(Met 17 sulfone), retained full activity. These structure-activity relationships for relaxation of brain arteries are consistent with previous studies of other biological responses to VIP.

Oral glucose inhibits growth hormone secretion induced by human pancreatic growth hormone releasing factor 1-44 in normal man.

The interaction between the inhibitory effect on growth hormone secretion of a 75 g oral glucose load and the stimulatory effect of human pancreatic growth hormone releasing factor 1-44 (hpGRF 1-44, 10 micrograms i.v.) has been studied in six normal subjects. hpGRF 1-44 alone induced a rise in growth hormone concentrations (maximum mean +/- SEM, 16.5 +/- 1.7 mU/l 15 min after injection) while growth hormone levels were suppressed by oral glucose alone (less than 1.5 mU/l from 45 to 135 min after glucose ingestion). When hpGRF 1-44 was injected 60 min after oral glucose, the growth hormone response was attenuated (maximum, 6.7 +/- 1.4 mU/l at 15 min, P less than 0.05). Increments of blood glucose within the physiological range diminish the growth hormone response to hpGRF 1-44 in normal man.

Cerebral Vascular Adenylate Cyclase: Evidence for Coupling to Receptors for Vasoactive Intestinal Peptide and Parathyroid Hormone

We have studied the responsiveness of vascular adenylate cyclase to vasoactive intestinal peptide (VIP) and parathyroid hormone (PTH) using preparations of cerebral microvessels and arteries. Cerebral microvessels obtained from rats, guinea-pigs, cattle, and pigs all responded potently to bovine (b) PTH-(1-34), whereas considerable between-species variability was observed in the responsiveness to VIP. The homologous peptide to VIP, PHI (porcine heptacosapeptide), stimulated adenylate cyclase in both rat microvessels and a broken-cell preparation of bovine arteries. The ED50 values for activation of bovine arterial adenylate cyclase by VIP, PHI, and bPTH-(1-34) were 6.9 nM, 10 nM, and 100 nM, respectively, with the following order of efficacy: VIP = PHI greater than bPTH-(1-34). The other related peptides, hpGRF (human pancreatic growth hormone releasing factor), secretin, and glucagon, and the fragment VIP-(10-28) were inactive. The PTH antagonist, [Nle8, Nle18, Tyr34]bPTH-(3-34) amide, inhibited bPTH-(1-34) activation of vascular adenylate cyclase but did not affect activation by VIP using either microvessels or arteries. VIP or PHI demonstrated an additive effect with bPTH-(1-34) on vascular adenylate cyclase activity. However, the effects of VIP and PHI were nonadditive with each other. These data suggest that VIP and bPTH-(1-34) activate cerebral vascular adenylate cyclase by interacting with pharmacologically distinct receptors, whereas PHI and VIP likely interact with a common receptor.

Plasma growth hormone responses to constant infusions of human pancreatic growth hormone releasing factor. Intermittent secretion or response attenuation.

Administration of human pancreatic tumor growth hormone (GH) releasing factor (hpGRF[1-40]) as a single injection to normal human subjects stimulates the secretion of GH in a dose-responsive manner. In the present studies, hpGRF(1-40) was infused in a graded stepwise manner over a 6-h period in order to determine whether the GH secretory response would be sustained. Normal adult males received four consecutive 90-min infusions of hpGRF(1-40) at doses of 1, 3.3, 10, and 33 ng/kg per min, preceded and followed by a 90-min saline infusion; and the plasma GH responses were compared with those during a separate control infusion. Plasma GH levels were significantly elevated by each hpGRF(1-40) infusion; and dose responsiveness was evident for the lowest three doses. Mean integrated GH secretory rates for the four doses were 1.95, 3.29, 4.29, and 3.65 times those of the respective control study. Plasma GH responses exhibited considerable variability, frequently decreasing during the latter part of each infusion; and at the highest dose, they decreased continuously beginning shortly after the onset of infusion. Episodic GH secretion occurred in individual subjects during each of the infusion periods. The possible contribution of hypothalamic somatostatin secretion to the diminished GH responsiveness was evaluated by determining plasma thyroid stimulating hormone (TSH) levels during the infusions and the TSH responses to thyrotropin-releasing hormone (500 micrograms i.v.) during a separate hpGRF(1-40) infusion of 2 ng/kg per min. Neither basal nor stimulated TSH levels differed between GRF-infused and control groups. The results indicate that GH secretion is dose responsive to hpGRF(1-40) infusions, though the response to hpGRF(1-40) infusions, though the response is complex. The absence of impaired TSH secretion provides evidence against a mediating role of somatostatin. The explanation for the loss of GH responsiveness remains undetermined but could include GRF-induced receptor down-regulation, a postreceptor effect, or, in spite of our negative results, a somatostatin-mediated inhibition.

Stimulation of growth hormone release in dwarf and normal chickens by thyrotrophin releasing hormone (TRH) or human pancreatic growth hormone releasing factor (hpGRF)

Stimulation of growth hormone release in dwarf and normal chickens by thyrotrophin releasing hormone (TRH) or human pancreatic growth hormone releasing factor (hpGRF)

Synthetic human pancreatic growth hormone releasing factor (hpGRF) stimulates growth hormone (GH) secretion [formula omitted] in chickens

Synthetic human pancreatic growth hormone releasing factor (hpGRF) stimulates growth hormone (GH) secretion [formula omitted] in chickens

Metabolic clearance and plasma disappearance rates of human pancreatic tumor growth hormone releasing factor in man.

The metabolic clearance rate (MCR) and plasma disappearance rate (t1/2) of human pancreatic tumor growth hormone releasing factor [hpGRF(1-40)] was determined in normal adult male subjects by single injection and constant infusion techniques. Single injections of 1, 3.3, and 10 micrograms/kg hpGRF(1-40) were administered intravenously, plasma immunoreactive (IR) GRF levels were measured during the subsequent 180 min, and biexponential curve analysis was performed. Graded, dose-constant infusions of hpGRF(1-40) at rates of 1, 3.3, 10, and 33 ng/kg per min were administered and the MCR was calculated from measurement of steady state plasma IR-GRF levels at each infusion rate. The postinfusion disappearance rate was determined by linear regression analysis of plasma IR-GRF levels during the 120-min period after cessation of the infusion. The calculated MCR during the single injection study was 194 +/- 17.5 liters/m2 per d and was not significantly different from the calculated value during the constant infusion study (202 +/- 16 liters/m2 per d). The disappearance rate during the single injection study was subdivided into two linear phases: an initial equilibration phase (7.6 +/- 1.2 min) and a subsequent elimination phase (51.8 +/- 5.4 min). The latter was similar to the linear disappearance rate observed (41.3 +/- 3.0 min) after cessation of the constant infusion. The chromatographic and biologic characteristics of plasma IR-GRF, 30 min after injection, were similar to those of synthetic hpGRF(1-40). The results have been discussed in relation to the MCR of other hypothalamic hormones and have been used to extrapolate secretion rates of GRF in patients with ectopic GRF production.

The effect of plasma glucose on the growth hormone response to human pancreatic growth hormone releasing factor in normal subjects.

Pituitary responsiveness to 44 amino acid human pancreatic growth hormone releasing factor was tested under conditions of euglycaemia and hyperglycaemia in six normal subjects. A 100 micrograms dose of growth hormone releasing factor was given at a fasting blood glucose of 5.1 +/- 0.4 mmols/l (mean +/- S.D.), and at a blood glucose level of 10.9 +/- 1.5. Under conditions of hyperglycaemia, the GH response to releasing factor was significantly depressed when compared to results obtained at fasting blood glucose (n = 6, t = 3.902, P = 0.0114). This is in keeping with the hypothesis that hyperglycaemia, mediated by the hypothalamus, causes decreased pituitary sensitivity to natural growth hormone releasing hormone.

Growth hormone releasing factor immunoreactivity in rat hypothalamus

Neurones immunoreactive to antibodies against human pancreatic growth hormone releasing factor 1–40 (hpGRF) were identified in the hypothalamus of the rat after pretreatment with colchicine. Reactive perikarya were concentrated in the arcuate nucleus and were also present around the anterior commissure. hpGRF immunoreactive fibres were observed in the median eminence and preoptic area where they tended to complement the distribution of somatostatin immunoreactive fibres. The distribution of GRF-immunoreactive perikarya in the rat hypothalamus is similar to that reported in monkey, and is consistent with other studies which suggest that neural mechanisms stimulatory for growth hormone secretion in the rat are situated in the medial basal hypothalamus.

Synthetic human pancreatic growth hormone releasing factor (GRF) stimulates growth hormone secretion in the domestic fowl ( [formula omitted

Synthetic human pancreatic Growth Hormone-Releasing Factor (hpGRF) elevated the plasma concentration of growth hormone (GH) in young and adult domestic fowl. This in vivo effect of hpGRF appeared to be largely similar for both the 32 amino-acid (hpGRF 1–32) or 40 amino-acid (hpGRF 1–40) polypeptide, although the effect of hpGRF 1–32 was more prolonged than that of hpGRF 1–40 in adult domestic fowl. The increase in plasma GH concentrations following hpGRF administration (10 μg/kg) was somewhat greater in young than adult chickens (the increase in plasma concentration of GH being 230 ng/ml at 1 week old, 282 ng/ml at 6 week old, 241 ng/ml at 10 weeks and 150 ng/ml in adults). In the adult domestic fowl hpGRF stimulated, a greater increase in the plasma concentration of GH than did thyrotropin-releasing hormone (TRH). However in the young chicks TRH was more active. The in vitro release of GH from dispersed chicken pituitary cells was elevated by hpGRF (1–32) and hpGRF (1–40).

Super-active analogs of growth hormone-releasing factor (1–29)-amide

Human pancreatic growth hormone releasing factor (1–29)-amide [hpGRF (1–29)-NH 2] and the following analogs: [D-Tyr-1]-hpGRF (1–29)-NH 2, [D-Ala-2]-hpGRF (1–29)-NH 2, [D-Asp-3]-hpGRF (1–29)-NH 2, and [N-Ac-Tyr-1]-hpGRF (1–29)-NH 2 were synthesized using solid phase methodology and tested for their ability to stimulate growth hormone (GH) secretion in the rat and the pig in vivo. [D-Ala-2]-hpGRF (1–29)-NH 2 was approximately 50 times more potent than the parent molecule in eliciting GH secretion in the rat. The other analogs were less active, but all were more potent than the 1–29 amide in the rat. [D-Tyr-1]-hpGRF (1–29)-NH 2 was 10 times more potent, [D-Asp-3]-hpGRF (1–29)-NH 2 7 times more potent, and the acetylated molecule approximately 12 times more potent than hpGRF (1–29)-NH 2.

Effects of growth hormone-releasing factor, somatostatin and dopamine on growth hormone and prolactin secretion from cultured ovine pituitary cells

A synthetic form of human pancreatic growth hormone releasing factor (GRF-44-NH 2) was shown to be a potent stimulator of growth hormone (GH) secretion and cellular cyclic AMP levels in cultured sheep pituitary cells. A small dose-dependent stimulation of prolactin secretion was also observed. Somatostatin (0.5 μM) completely blocked the maximal GRF (1 nM)-stimulated secretion without a significant effect on cyclic AMP levels. Dopamine (0.1 μM) inhibited the GRF-elevated GH secretion by 50% and lowered cyclic AMP levels by 30%. Dopamine (0.1 μM) inhibition of basal prolactin secretion was not affected by GRF (1 nM). The data support the hypothesis that cyclic AMP is involved in the action of GRF but suggest that somatostatin can inhibit GRF-induced secretion of GH independently of cyclic AMP.

Plasma growth hormone responses to repetitive administrations of growth hormone releasing factor in patients with pituitary dwarfism.

Plasma growth hormone (GH) responses to the repetitive administrations of synthetic human pancreatic growth hormone releasing factor (hpGRF-44) were studied in 15 patients with GH deficiency (11 diagnosed as idiopathic and 4 diagnosed as secondary to hypothalamo-pituitary tumor). hpGRF-44 was administered by single iv bolus (2 micrograms/kg), repetitive im (100 micrograms, twice a day), and/or repetitive iv infusion (2.5 micrograms/min for 90 min, once a day) for three to six consecutive days. Three of the eleven idiopathic GH deficient patients had plasma GH responses to both single iv bolus injection and repetitive administrations by im, or iv infusion of hpGRF. In four of the remaining eight, who had not had peak plasma GH levels above 5 ng/ml to a single iv bolus of the peptide, repetitive administrations of hpGRF-44 by im injection and/or iv infusion induced GH responses to the peptide. In the four patients with secondary GH deficiency, three had plasma GH response to hpGRF administration but one patient, who had indications of pituitary disorder, did not show any plasma GH response to either single iv injection or repetitive administrations of hpGRF-44. These data show that repetitive administrations of hpGRF-44 can induce plasma GH responses in some GH deficient patients who do not respond to a single iv bolus of the peptide.

Growth hormone and prolactin response to thyrotropin releasing hormone and growth hormone releasing factor in the immature turkey.

Synthetic thyrotropin releasing hormone (TRH) and human pancreatic growth hormone releasing factor (hpGRF) stimulated growth hormone (GH) secretion in 6- to 9-week-old turkeys in a dose-related manner. TRH and hpGRF (1 and 10 micrograms/kg, respectively) each produced a sixfold increase in circulating GH levels 10 min after iv injection. Neither TRH nor hpGRF caused a substantial change in prolactin (PRL) secretion in unrestrained turkeys sampled through intraatrial cannulas. However, some significant increases in PRL levels, possibly related to stress, were noted.

Growth hormone releasing factors in the brain and the gut: Chemistry, actions, and localization

Within the physiological range of other known releasing factors, human pancreatic tumor growth hormone releasing factor (hpGRF) is specific for GH release. Data concerning hpGRF action on cAMP and GH are consistent with the concept of cAMP acting as a second messenger for this releasing factor. hpGRF-stimulated GH release is Ca ++ dependent. Exogenous hpGRF 40 does not alter the interdigestive gastric motility or secretion of gastrin and motilin in dogs, while large doses of hpGRF stimulate somatostatin release into the hepatic portal blood of the rat. Significant GRF activity as determined by a rat pituitary perifusion system is confined within the median eminence and the arcuate nucleus, though detectable but insignificant GRF activity is present in other area of the hypothalamus and cortex in the rat. GRF activity is present in the ovine brain as well as in the gut. Both tissues contain large (between 4000–5000 daltons) and small (but possibly larger than 1000 daltons) m.w. GRF materials. GRF appears to be structurally different between species and more than one GRF may be present within the same species. One of the ovine brain peptides with GH-releasing activity was partially characterized as His-Ser-Asp-Gly-Ile-Phe-Thr-Asp-Ser-Tyr-Lys-Arg-Try-Asn-Lys-Glu-Met-Ala-Lys—which is similar to rat GRF and porcine VIP having His at the N-terminus. Another peptide with GRF activity which eluted earlier on reverse phase HPLC and later on cation exchange chromatography has also been obtained in a pure form. In addition, VIP which has low but significant GRF activity in vitro may potentiate the GRF action of hpGRF in vivo and in vitro under certain conditions, suggesting a potentially physiologically important interplay of VIP and GRF with respect to GH secretion.

Reversed-phase high-performance liquid chromatography: preparative purification of synthetic peptides

Biologically active peptides synthesized by the solid phase methodology of Merrifield were purified by reversed-phase high-performance liquid chromatography using newly developed preparative radially compressed cartridges fitting Waters Assoc. Prep LC 500 liquid chromatograph. Cartridges were handpacked with Vydac C 18, C 4 or diphenyl derivatized silicas (pore size 300 Å) of different particle sizes (10–20 μm). Large scale purification of gram amounts of gonadotropin releasing hormone analogs (agonist and antagonist) as well as amidated human pancreatic tumor growth hormone releasing factor (a 40-peptide) illustrate the resolutive power of this technique applied to the isolation of more than 300 synthetic peptides in our laboratory over the last two years. Difficult separations were achieved by changing supports (C 18, C 4, diphenyl) as well as mobile phase composition: (triethylammonium phosphate pH 2.25 or 6.5, 0.1% trifluoroacetic acid, ammonium acetate pH 6.5 and acetonitrile). Protected amino acids and peptides amenable to normal-phase chromatography on Vydac spherical underivatized silica were purified economically by the reversed-phase mode. It is understood that this general, convenient and versatile strategy may be applicable to the preparative scale isolation of any other class of compounds usually separated on reversed-phase high-performance liquid chromatography.

In vivo and in vitro stimulation of growth hormone release in chickens by synthetic human pancreatic growth hormone releasing factor (hpGRFs).

The effect of the synthetic human pancreatic GH releasing factors (hpGRFs, hpGRF44 and hpGRF40) on GH release was studied both in vivo and in vitro. Four-week-old cockerels were injected iv with hpGRFs at 0.1 microgram, 1.0 microgram, and 10.0 micrograms/bird. The 0.1 microgram dose of hpGRFs had no effect on plasma GH levels, but the 1.0 microgram and 10.0 micrograms doses of hpGRFs caused a dose-dependent elevation of plasma GH. Plasma GH levels returned to basal 60 min after injection. HpGRFs, thyrotropin releasing hormone (TRH) and somatostatin (SRIF) were examined in a chicken in vitro pituitary cell culture system. In vitro hpGRFs and TRH caused a significant dose-dependent release of chicken GH, and the ability to stimulate GH was additive when hpGRF44 and TRH were added together in the culture medium. SRIF showed no consistent effect on the release of chicken GH, but it inhibited the stimulatory effect of hpGRF44 on chicken GH release. We conclude that hpGRF is a potent releaser of chicken GH in vivo and in vitro.