Naltrexone reduces weight gain, alters “β-endorphin”, and reduces insulin output from pancreatic islets of genetically obese mice

Naltrexone, an opiate antagonist, was administered to young obese (ob/ob) and lean mice for five weeks. Animals had continuous access to food and received 10 mg/kg SC twice daily with equivalent volumes of saline given to controls. The effects on body weight, and pituitary and plasma levels of β-endorphin-like material were measured. Naltrexone-injected obese animals gained weight more slowly over the first three weeks while the weight gain of lean animals was not affected by naltrexone. Plasma levels of β-endorphin were shown to be significantly higher in untreated ob/ob mice and this difference increased with age (4–20 weeks). With naltrexone treatment, plasma levels in +/? mice rose and exceeded those in ob/ob. Saline treatment appeared to be a stress, and pituitary β-endorphins rose 4–6 fold in ob/ob compared with +/?. While naltrexone reduced the levels in ob/ob pituitary towards normal, no effect on β-endorphin levels in pituitary of lean mice was obtained. In vitro studies of effects of the opiate antagonists, naloxone, on insulin secretion by isolated islets provided additional evidence of resistance of lean mice to naloxone relative to ob/ob. (IRI secretion fell only in naloxone treated ob/ob islets.) These observations support the contention that this form of genetic obesity is characterized by elevated endogenous opiate levels and an increased sensitivity to opiate antagonists such as naltrexone or naloxone.     

Effect of beta-endorphin on gastric acid secretion and serum gastrin concentration in humans

The effect of synthetic human β-endorphin on gastric acid secretion was studied in 9 healthy subjects. Neither 2.5 mg or 15 mg β-endorphin had a significant effect on acid secretion or on serum gastrin concentration despite the fact that this dose of opiate caused a significant increase in serum prolactin concentrations. The role of endogenous opiate-like peptides on gastric secretion is discussed.     

Naltrexone reduces weight gain, alters “β-endorphin”, and reduces insulin output from pancreatic islets of genetically obese mice

Naltrexone, an opiate antagonist, was administered to young obese (ob/ob) and lean mice for five weeks. Animals had continuous access to food and received 10 mg/kg SC twice daily with equivalent volumes of saline given to controls. The effects on body weight, and pituitary and plasma levels of β-endorphin-like material were measured. Naltrexone-injected obese animals gained weight more slowly over the first three weeks while the weight gain of lean animals was not affected by naltrexone. Plasma levels of β-endorphin were shown to be significantly higher in untreated ob/ob mice and this difference increased with age (4–20 weeks). With naltrexone treatment, plasma levels in +/? mice rose and exceeded those in ob/ob. Saline treatment appeared to be a stress, and pituitary β-endorphins rose 4–6 fold in ob/ob compared with +/?. While naltrexone reduced the levels in ob/ob pituitary towards normal, no effect on β-endorphin levels in pituitary of lean mice was obtained. In vitro studies of effects of the opiate antagonists, naloxone, on insulin secretion by isolated islets provided additional evidence of resistance of lean mice to naloxone relative to ob/ob. (IRI secretion fell only in naloxone treated ob/ob islets.) These observations support the contention that this form of genetic obesity is characterized by elevated endogenous opiate levels and an increased sensitivity to opiate antagonists such as naltrexone or naloxone.     

The roles of the opioid peptides in controlling thyroid stimulating hormone release

We have studied the role of the opioid peptides in controlling TSH secretion. Morphine sulfate significantly decreased, while naloxone had no effect on, basal plasma TSH levels of female rats. In contrast, naloxone blocked the stress-induced fall in plasma TSH. Microinjection of β-endorphin into the third ventricle resulted in a fall in TSH while such injection of naloxone into the posterior hypothalamus increased TSH. Microinjection of β-endorphin directly into the pituitary caused a rise in plasma TSH. It is concluded that opioid peptides probably play no role in basal TSH secretion, but are involved in the stress-induced fall in TSH. Furthermore, it appears that opioid peptides have a site of action in the hypothalamus to decrease TSH and a direct pituitary action to increase TSH.     

Suppression of serum prolactin by naloxone but not anti-β-endorphin antiserum in stressed and unstressed rats

The role of endogenous opioids in the regulation of tonic and stress-induced prolactin secretion was studied in male rats. Animals with chronically indwelling intra-atrial catheters were used and served as their own controls. Intravenous injection of a potent rabbit anti-β-endorphin antiserum produced no change in either baseline serum prolactin or in the rise induced by swimming for 15 minutes at 20 C. Naloxone, 0.5 mg/kg intravenously, produced a small but statistically significant lowering of baseline serum prolactin levels, as well as a mild blunting of the stress-induced serum prolactin rise. The failure of anti- - endorphin antiserum to affect serum prolactin may be explained either by failure of the antiserum to gain access to hypothalamic prolactin regulating loci, or to lack of involvement of β-endorphin in the control of serum prolactin. The suppression of prolactin secretion seen with naloxone indicates that endogeneous opioids are involved in prolactin regulation, though the relatively small magnitude of the changes observed suggests that their role under physiological conditions may be a minor one.     

Effects of beta-endorphin, met-enkephalin and somatostatin on the neurotensin-induced neurogenic contraction in the guinea-pig ileum

At maximally effective concentrations, the opiate peptides β-endorphin (240 nm) and Met-enkephalin (1400 nM) virtually abolished the contractions induced by a maximally effective concentration of 60 nM neurotensin (NT), either in the longitudinal smooth muscle strip or in the intact segment of guinea-pig ileum. This inhibitory effect was concentration-dependent and was totally blocked by naloxone at 100 nM. In contrast a maximally effective concentration of somatostatin (60 nM) partially inhibited (50–60%) the contraction induced by 60 nM NT in either smooth muscle preparation. Somatostatin inhibition was concentration-dependent and was not blocked by naloxone at 100 nM. Atropine at 100 nM inhibited by 50% the contractions induced by 60 nM NT in the intact segment of guinea-pig ileum. The remaining contraction was abolished by β-endorphin and Met-enkephalin and partially reduced by somatostatin. Our results confirm that NT-induced contractions in the guinea-pig ileum are neurogenic and involve a cholinergic as well as a non-cholinergic component. Furthermore, we show that the release of mediators from both components     

Alpha-endorphin, gamma-endorphin and their des-tyrosine fragments in rat pituitary and brain tissue

Enkephalins, endorphins and related peptides were determined in pituitary and brain tissue of rats which were killed by decapitation or microwave irradiation. The tissues were heated in 1M acetic acid prior to homogenization and the levels of the various peptides were measured by means of a combination of HPLC and radioimmunoassays. Enkephalin levels in pituitary and brain of irradiation-killed rats were much higher as compared to those in tissue of rats sacrificed by decapitation. Similar data were obtained with respect to pituitary levels of γ-endorphin, des-Tyr-γ-endorphin and des- Tyr-α-endorphin. However, brain levels of α- and γ-endorphin and their respective des-Tyr-fragments were not different with the two methods of sacrifice used. The concentrations of β-endorphin in the pituitary gland were similar in rats killed by microwave irradiation and decapitation, but irradiation showed higher β-endorphin levels in the brain than decapitation. These results suggest that β-endorphin fragments like α- and γ-endorphin and des-Tyr-α- and des-Tyr-γ-endorphin are endogenous peptides in the rat pituitary gland and the brain.     

Regulation of hypophysial secretion by endogenous opiates: Humoral endorphin stimulates the release of growth hormone

Humural endorphin, a recently discovered endogenous opioid factor stimulates the release of growth hormone and, to some extent of prolactin, similarly to other endogenous (enkephalin, β-endorphin) and exogenous (morphine) opiates. This stimulatory effect is dose-dependent with peak values at 30 minutes following intraventricular injection to newborn rats. However, in contrast to the other opioid ligands, the effect of humoral endorphin is not blocked in a dose-dependent fashion by naloxone, the potent opiate antagonist. Thus, while moderate doses of naloxone partially inhibit the stimulatory effect, higher doses which completely block morphine, enkephalin and β-endorphin, are ineffective in antagonizing humoral endorphin. This peculiar interaction between naloxone and humoral endorphin resembles the effect of the opiate antagonist on spontaneous release of growth hormone and prolactin, suggesting the involvement of humoral endorphin in the physiological regulation of hypophysial secretion.     

beta-Endorphin in human plasma: basal and pathologically elevated levels.

β-Endorphin-like immunoreactivity was measured in plasma of normal human subjects and in plasma of patients with pathologically elevated ACTH levels. The antiserum used displayed the same avidity for human β-endorphin and human β-lipotropin (detection limit for both peptides 1–2 fmoles/tube). Gel chromatography of the immunoreactive components in plasma of normal subjects indicated the presence of both β-lipotropin (2.1 -10.1 fmoles/ml) and β-endorphin (3.5–6.4 fmoles/ml). A close correlation between immunoreactive β-endorphin and ACTH was found in plasma of patients suffering from Addison's disease, Cushing's disease and exhibiting Nelson's syndrome. Elevated levels of β-endorphin-like immunoreactivity in plasma of these patients were due to both β-lipotropin and β-endorphin.     

β-endorphin: Effects on respiratory regulation

Intracisternal injection of 14.5 nmoles of human β-endorphin in lightly anesthetized dogs resulted in marked respiratory depression, manifested by diminished responses of ventilation and airway occlusion pressure to carbon dioxide rebreathing. These responses were temporarily reversed by intravenous injection of naloxone and attenuated following a second β-endorphin injection. Results in this study suggest a possible physiological role for endogenous opioid peptides in the regulation of respiration.     

The effect of β-endorphin on arginine-induced growth hormone and prolactin release

β-endorphin administration via constant infusion inhibited the release of growth hormone (GH) and augmented the release of prolactin (PRL) induced by arginine in normal female subjects. Although β-endorphin infusion also induced hyperglycemia, the increment in plasma glucose was insufficient to account for the observed suppression of arginine-initiated GH release. These studies demonstrate that β-endorphin influences, in opposed directions, the secretion of PRL and GH in women.     

Elevated plasma beta-endorphin levels in pregnant women and their neonates.

Using radioimmunoassay technique β-endorphin levels were measured in the plasma of women undergoing labour and partirition and in the plasma of their neonates. The level of immunoreactive β-endorphin in the plasma of women undergoing labour was found to be significantly elevated (mean values: 38–135 fmoles/ml) above the levels found in non-pregnant women (mean values: 5–10 fmoles/ml). After birth, the level of β-endorphin-like immunoreactivity in maternal venous plasma was significantly higher than that in the umbilical vein and artery plasma of the new-borns, but there was no arterio-venous difference in the neonatal plasma. Since the antiserum used displayed the same avidity for human β-endorphin and β-lipotropin chromatographic separation of the immunoreactive components was performed by gelfiltration. Both peptides were found in the plasma of non-pregnant women, in maternal plasma and in the plasma of the neonates. In addition, high amounts of both peptides were found in the fetal pituitary gland showing that the fetus can probably produce its own peptides.     

Endorphins and the central inhibition of urinary bladder motility

The involvement of endogenous opioid mechanisms in the central neurogenic control of urinary bladder function has been examined in anesthetized rats. Intracerebroventricular (ICV) microinjection of β-endorphin (0.5–2.0 μg) produced powerful inhibition of rhythmic bladder contractions initiated by central reflex activity. The peptide fragments γ-endorphin and α-endorphin (4–16 μg), formed by the processing of β-endorphin by membrane homogenates of brain, were less active than the parent compound. The inhibitory effects of β-endorphin was reversed by ICV naloxone (1–2 μg) but higher doses were required to reverse γ- or α-endorphin effects. ICV naloxone administered alone increased intravesicular pressure and bladder contraction frequency. These observations support the hypothesis that the endorphins have a physiological role in the central regulation of urinary bladder activity.     

Neonatal administration of beta-endorphin produces "chronic" insensitivity to thermal stimuli

Male and female rat pups were injected with β-endorphin, naloxone or a saline control solution during days 2–7 postnatally. At 90 days of age the rats were tested for analgesia with the tail flick test. Testing was conducted during the first 2 hours of the light and the dark cycle. In both sexes and during both phases of the light cycle rats treated with β-endorphin as infants evidenced a significant elevation in threshold for painful thermal stimuli. Early treatment with naloxone also resulted in elevated threshold for thermal stimuli. Administration of naloxone to these rats as adults did not reverse the analgesic effect. It was concluded that early exposure to β-endorphin results in permanent changes in behavior perhaps by altering the interaction of endogenous opiates with their binding sites during a ciritcal period of opiate receptor development.     

Effects of peptides of pituitary origin on the formation of C21 steroid by fetal calf adrenal cells in culture.

Corticotrophic activity of opiate-like peptides was assessed by their ability to stimulate the formation of C₂₁ steroids from [³H] progesterone by three-day old cultures of fetal calf adrenal cells. ACTH_1–39, ACTH_α1–24 and a purified preparation of pituitary ovine β-endorphin caused a marked increase in 17α and 21-hydroxylation while a preparation of pure synthetic porcine β-endorphin gave a minimal stimulation. The activity of the purified ovine β-endorphin preparation could not be accounted for by contamination by ACTH or by a synergistic action between the two peptides. The novel pituitary factor described here may be due to a contaminant of the β-endorphin peak which is different from ACTH_1–39.     

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

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

Growth hormone secretion during sleep. II. Interrelationships between growth hormone secretion, insulin-like growth factor I and sex steroids

The serum levels of insulin-like growth factor I (IGF I), dehydroepiandrosterone sulfate (DHAS), testosterone (T) and estradiol (E2) have been measured in 78 prepubertal and 57 early pubertal patients referred for short stature, at the same time when their secretion of GH was evaluated both during nocturnal sleep and by two conventional stimulation tests. According to the results of GH measurements they were considered as having a normal secretion of GH (group I), a complete GH deficiency (group II), a partial GH deficiency (group III), low responses to stimuli with normal secretion during sleep (group IV) or a nocturnal neurosecretory dysfunction (group V). Though widely scattered, the IGF I levels showed the following characteristics: a significant increase at puberty from 0.77 to 1.29 U/ml (p less than 0.001) in the so-called endocrinologically normal patients of group I, not in the other groups; in the prepubertal patients of group I, a correlation of IGF I with chronological age (r = 0.47, p less than 0.005) and bone age (r = 0.52, p less than 0.002); significantly reduced IGF I levels in patients of group II having complete GH deficiency (p less than 0.001); no significant differences between prepubertal patients with partial or atypical GH deficiency from groups III, IV, V and prepubertal patients from group I; lower pubertal levels in groups III, IV, V than in pubertal patients from group I (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Studies of naloxone-induced secretion of β-endorphin immunoreactivity in dogs

The stimulating effect of naloxone on plasma β-endorphin immunoreactivity (βEI) was examined in dogs. Intravenous naloxone at 5, 1, or 0.1 mg/kg caused a significant increase in βEI while doses of 0.01 or 0.001 mg/kg had no effect. The peak plasma βEI levels occurred at 25 mins after naloxone. The neurotransmitter and antagonists metergoline, atropine, diphenhydramine and phentolamine all failed to significantly alter basal βEI secretion; further, they all failed to prevent the increase in βEI resulting from naloxone administration. Dexamethasone prevented the naloxone-induced rise in βEI. Our results suggest naloxone's effect on βEI is not mediated through several neurotransmitter systems known to affect ACTH secretion. Additionally, βEI secreted in response to naloxone appears to originate mainly from the anterior lobe of the pituitary.     

Morphine treatment selectively regulates expression of rat pituitary POMC and the prohormone convertases PC1/3 and PC2

The prohormone convertases, PC1/3 and PC2 are thought to be responsible for the activation of many prohormones through processing including the endogenous opioid peptides. We propose that maintenance of hormonal homeostasis can be achieved, in part, via alterations in levels of these enzymes that control the ratio of active hormone to prohormone. In order to test the hypothesis that exogenous opioids regulate the endogenous opioid system and the enzymes responsible for their biosynthesis, we studied the effect of short-term morphine or naltrexone treatment on pituitary PC1/3 and PC2 as well as on the level of pro-opiomelanocortin (POMC), the precursor gene for the biosynthesis of the endogenous opioid peptide, β-endorphin. Using ribonuclease protection assays, we observed that morphine down-regulated and naltrexone up-regulated rat pituitary PC1/3 and PC2 mRNA. Immunofluorescence and Western blot analysis confirmed that the protein levels changed in parallel with the changes in mRNA levels and were accompanied by changes in the levels of phosphorylated cyclic-AMP response element binding protein. We propose that the alterations of the prohormone processing system may be a compensatory mechanism in response to an exogenous opioid ligand whereby the organism tries to restore its homeostatic hormonal milieu following exposure to the opioid, possibly by regulating the levels of multiple endogenous opioid peptides and other neuropeptides in concert.