Inhibitory action of the alpha 1-adrenergic receptor on growth hormone secretion in the dog.

Intravenous administration of methoxamine ( METHOX , 5 mg, iv), a specific alpha 1-adrenergic agonist, reduced baseline GH levels of four unanesthetized beagle dogs 45 and 60 min post-injection and completely abolished the GH-releasing effect of the alpha 2-adrenergic agonist clonidine ( CLON , 4/ micrograms/kg, iv). Prazosin (PRA, 0.1 mg/kg, iv), an alpha 1-adrenergic antagonist, administered before METHOX re-instituted the GH-releasing effect of CLON . METHOX administered at the starting of an arginine infusion (ARG, 10% solution, 3.3 ml/min X 30 min) reduced consistently the GH releasing effect of the latter while, conversely, pretreatment with PRA, strikingly potentiated the GH-releasing effect of the amino acid (2 dogs). METHOX did not alter the GH-releasing effect of human pancreatic GH-releasing factor (hpGRF-40, 1/microgram/kg, iv), though it consistently delayed the occurrence of the GH secretory peak following hpGRF-40. These data indicate that alpha 1-adrenergic receptors located in the central nervous system, inhibit in the dog tonic and stimulated GH secretion.

Opiates act centrally on GH and PRL release.

This study was conducted in order to further understand whether the effect of opiates on growth hormone and prolactin release was exerted centrally or at least in part peripherally. Male rats were treated with morphine-HCl after pretreatment with saline with either the opiate antagonist naloxone-HCl or the quaternary derivative naloxone methyl bromide (Naloxone-Br), the latter of which does not cross the blood brain barrier. Morphine-HCl elicited a clear cut increase in prolactin and growth hormone release after pretreatment with naloxone-Br, but not after pretreatment with naloxone-HCl. Naloxone-Br, however, was able to inhibit the effect of morphine when administered directly in the brain ventricles. To further confirm these results, we administered the quaternary derivative morphine-methyl-iodide (morphine-I), which unlike morphine-HCl, does not cross the blood brain barrier. Morphine-I was ineffective in eliciting growth hormone and prolactin release when administered peripherally, but was effective when administered intraventricularly.

Proof for histaminergic but not for adrenergic involvement in the growth hormone-releasing effect of an enkephalin analog in the dog.

A series of studies was performed in unanesthetized dogs to ascertain whether, in addition to cholinergic pathways, other neurotransmitter systems were involved in the GH-releasing effect of the potent enkephalin analog [D-Ala2, MePhe4, Met(o)5-ol]enkephalin (DAMME). DAMME at a dose of 8 microgram/kg iv elicited a striking rise in plasma canine GH (cGH), with peak levels at 30 min. Blockade of histaminergic H1 receptors by diphenhydramine (30 mg,iv, 15 min before) or clemastine (1 mg orally three times for 2 days and 2 mg orally 60 min before) completely suppressed the cGH release induced by DAMME without significantly altering baseline cGH levels. A slight reduction of the effect of DAMME was also induced by the histamine H2 receptor antagonist cimetidine (300 mg, iv, 15 min before). Pretreatment with the alpha-adrenergic inhibitor phentolamine (0.4 mg/min for 45 min) did not alter the neuroendocrine effect of DAMME, despite the occurrence of a rise in blood glucose (peak levels, 185 +/- 47 mg/dl). The administration of propranolol, a blocker of beta-receptors, did not potentiate the cGH release induced by a threshold dose of DAMME (4 microgram/kg, iv). An iv bolus injection of glucose (1 g/kg), which induced peak glucose levels of 296 +/- 29 mg/dl, completely suppressed the cGH release induced by DAMME or propranolol plus DAMME. These results indicate that histaminergic H1 receptors play an important role in the cGH release induced by DAMME, whereas this action occurs independently from adrenergic mediation. Based on these and previous findings, a neuromodulator role in GH-releasing mechanisms is suggested for opioid peptides.

Carbidopa inhibits the growth hormone- and prolactin-suppressive effect of L-dopa in acromegalic patients.

The plasma GH, PRL, TSH, and dopamine (DA) responses to an infusion of L-dopa were examined in six acromegalic patients before and after pretreatment with carbidopa, a drug which inhibits the peripheral conversion of L-dopa to DA. Carbidopa neither modified baseline DA nor induced changes in baseline GH, PRL, or TSH levels. The drug instead markedly reduced the L-dopa-induced DA rise, an effect which was concomitant to a striking reduction of the suppressive effect of L-dopa on plasma GH and a partial inhibition of the suppressive effect of L-dopa on plasma PRL. TSH levels did not change either after L-dopa alone or L-dopa plus carbidopa. These data demonstrate that in "responder" acromegalics, L-dopa inhibits GH secretion through its peripheral conversion to DA and not via activation of central DA neurotransmission. For the effect of L-dopa on PRL secretion, in addition to a peripheral dopaminergic component, a central component cannot be disregarded.

Plasma beta-endorphin, beta-lipotropin, and met-enkephalin concentrations during pregnancy in normal and drug-addicted women and their newborn.

Most studies of plasma beta-endorphin concentrations in pregnant women show that these are highly elevated. This might indicate a role for opiate peptides during pregnancy and in the fetus-mother relationship. We measured plasma beta-endorphin, beta-lipotropin, and met-enkephalin concentrations in normal and drug-addicted women during pregnancy, labor, and delivery, and in their newborn infants. Peptides were measured by RIA after extraction and concentration on silica columns and separation by high pressure liquid chromatography. In both normal and drug-addicted mothers we found an increase in plasma beta-endorphin during pregnancy, without a concomitant increase in plasma beta-lipotropin or metenkephalin. Only beta-lipotropin increased dramatically in both groups at delivery, whereas beta-endorphin and met-enkephalin remained unchanged. Peptide concentrations in umbilical plasma were similar to those in peripheral plasma of the mothers. On day 1 of life plasma beta-endorphin, beta-lipotropin, and met-enkephalin concentrations in the newborn from normal mothers were higher than in nonpregnant adult subjects and gradually decreased toward normal adult values by day 5 of life. Plasma beta-endorphin, beta-lipotropin, and met-enkephalin concentrations of newborn infants of drug-addicted mothers increased dramatically on day 2 and 3 of life, up to 1000-fold the concentrations of normal adults, and remained elevated up to 40 days after birth. In conclusion, beta-endorphin, beta-lipotropin, and met-enkephalin concentrations during pregnancy are not affected by drug addiction, whereas in the newborn of drug addicted mothers concentrations of these compounds are markedly increased.

Epidermal growth factor-induced phosphoinositide hydrolysis. Modulation by protein kinase C.

A short-term treatment with phorbol 12,13-dibutyrate (PDBu) was found to inhibit totally the epidermal growth factor (EGF)-stimulated phosphoinositide hydrolysis in A431 cells, whereas long-term pretreatment with PDBu, which is known to down regulate protein kinase C, induced a greater accumulation of the EGF-triggered inositol phosphate accumulation, particularly of Ins(1,3,4,5)P4. The increased Ins(1,4,5)P3/Ins(1,3,4,5)P4 formation in the PDBu long-term pretreated cells was coincident with the increased Ca2+ influx stimulated by EGF in the same cells. Since long-term pretreatment with PDBu was found to enhance the EGF signals, an explanation for the synergism between EGF and phorbol esters in the induction of DNA synthesis is provided.

The cost-effectiveness of multiple sclerosis rehabilitation: a model.

Many patients with multiple sclerosis (MS) who have fixed, chronic neurologic deficits are kept at home with the help of community support systems, which cannot improve the physical independence of the MS patients they serve. We have found that active, intensive, multidisciplinary rehabilitation effectively improves functional performance, even though it cannot alter the underlying disorder. Inpatient rehabilitation with follow-up home care may actually be less expensive than any other current health care alternative.

Effect of aspirin on serotonin and met-enkephalin in brain: correlation with the antinociceptive activity of the drug.

Intravenous administration of acetyl salicylate of lysine, a soluble salt of aspirin, reduced in rats the firing discharge of thalamic neurones, evoked by noxious stimuli. Concomitantly, concentrations of 5-hydroxyindole acetic acid increased, while those of met-enkephalin-like immuno-reactive derivatives were decreased in several areas of the brain. Similar electrophysiological and biochemical responses were obtained by administering tryptophan or 5-hydroxytryptophan plus carbidopa. The effect of aspirin on the evoked firing of the thalamic neurones was counteracted by pretreating the animals with metergoline. On the other hand, naloxone did not antagonize the inhibitory effect of aspirin and 5-hydroxytryptophan on pain-induced neuronal excitation. These data indicate that a serotonin-, but not a naloxone-sensitive opiate mechanism, may be relevant for aspirin-mediated antinociception.

Effect of a cholecystokinin antagonist on some effects of diazepam.

Dose responses were evaluated for the effects of diazepam alone or together with the cholecystokinin receptor antagonist CR 1409 on pentetrazole-induced convulsions, motor performance and spontaneous motor activity. The results obtained showed that the cholecytokinin antagonist potentiated the effects of diazepam on motor performance and the anticonvulsant activity of diazepam, while it did not affect spontaneous motor activity. The data presented are consistent with a role for cholecystokinin in some effects of diazepam.

Tolerance to some behavioural effects of lisuride, a dopamine receptor agonist, and reverse tolerance to others, after repeated administration.

To investigate whether prolonged pretreatment with the dopamine (DA) agonist lisuride would result in modification of some of its behavioural effects, food intake, locomotor activity, body temperature or stereotyped and mounting behaviour were evaluated after acute injections of different doses of lisuride into rats, pretreated daily for four weeks with either saline or lisuride. Rats pretreated with lisuride developed tolerance to its anorexigenic and hypothermic effects, and reverse tolerance to its effects on locomotor activity, stereotyped and mounting behaviour. Pretreatment with lisuride did not modify the activity of drug-metabolizing enzymes in the liver. These results, in addition to revealing the pattern of the changes in the behavioural effects of a DA agonist drug, after repeated administration, may be taken as evidence for the existence of different DA receptor systems in different areas of the brain, that mediate different behavioural effects, and that differ markedly in their reactions to prolonged stimulation with an agonist drug.

Pharmacological modulation of neuropeptides in peripheral mononuclear cells.

The concentrations of beta-endorphin and cholecystokinin were measured in fresh resting peripheral mononuclear cells obtained from rats and human subjects in basal conditions and after different pharmacological treatments. Both in the human and the rat, beta-endorphin concentrations in mononuclear cells, increased after treatment with serotoninergic agonists, decreased after dopaminergic or GABAergic drugs, while the respective antagonists exerted the opposite effect. In vitro, serotoninergic and GABAergic compounds confirmed their roles in the modulation of beta-endorphin in mononuclear cells. Cholecystokinin was never affected by the pharmacological treatments.

Antinociceptive and immunosuppressive effects of opiate drugs: a structure-related activity study.

1. Although it is well known that morphine induces significant immunosuppression, the potential immunosuppressive activity of morphine derived drugs commonly used in the treatment of pain (codeine, hydromorphone, oxycodone) has never been evaluated. 2. We evaluated in the mouse the effect of the natural opiates (morphine and codeine) and synthetic derivatives (hydromorphone, oxycodone, nalorphine, naloxone and naltrexone) on antinociceptive thresholds and immune parameters (splenocyte proliferation, Natural Killer (NK) cell activity and interleukin-2 (IL-2) production). 3. Morphine displayed a potent immunosuppressive effect that was not dose-related to the antinociceptive effect, codeine possessed a weak antinociceptive effect and limited immunosuppressive activity; nalorphine, a mu-antagonist and kappa-agonist, exerted a potent immunosuppressive effect, but had very weak antinociceptive activity. The pure kappa-antagonist nor-BNI antagonized the antinociceptive, but not the immunosuppressive effect of nalorphine. 4. Hydromorphone and oxycodone, potent antinociceptive drugs, were devoid of immunosuppressive effects. 5. The pure antagonists naloxone and naltrexone potentiated immune responses. 6. Our data indicate that the C6 carbonyl substitution, together with the presence of a C7-8 single bond potentiates the antinociceptive effect, but abolishes immunosuppression (hydromorphone and oxycodone). 7. The single substitution of an allyl on the piperidinic ring resulted in a molecule that antagonized the antinociceptive effect but maintained the immunosuppressive effect. 8. Molecules that carry modifications of C6, the C7-8 bond and C14, together with an allyl or caboxymethyl group on the piperidinic ring antagonized both the antinociceptive and the immunosuppressive effect of opiates and were themselves immunostimulants.

Purine involvement in morphine antinociception.

The effects of a series of adenosine derivatives on morphine antinoceptive effect were investigated in rats by the 'tail-flick' method. 2-Chloroadenosine (CADO) and L-N6-phenylisopropyladenosine (L-PIA), given intraperitoneally, caused decreased morphine antinociception. Intracerebroventricular injections of CADO, L-PIA and 5'-N-ethylcarboxamide adenosine (NECA), but not of 2'-deoxyadenosine, antagonized morphine antinociception. The effects of both central and peripheral injections of CADO and L-PIA on morphine antinociception were partially reversed by caffeine. Intracerebroventricular injection of dibutyryl-cyclic 3', 5' adenosine monophosphate (db cyclic AMP) had no effect on morphine antinociception. These data indicate that adenosine plays a role in morphine-induced antinociception. The results are discussed in terms of postulated effects of adenosine derivatives on adenylate cyclase.

The 5-hydroxytryptamine-like actions of 5,6-dihydroxytryptamine.

1. With isolated preparations of rat stomach fundus as well as of duodenum and ileum of rats and guinea-pigs, 5,6-dihydroxytryptamine and 5,6-diacetoxytryptamine caused a contraction which was antagonized by methysergide and lysergic acid diethylamide (LSD), but not by atropine. Pretreatment of the animals with reserpine did not decrease the effect of the two indoleamines on the isolated ileum and duodenum.2. In anaesthetized guinea-pigs, 5,6-dihydroxytryptamine and its diacetyl derivative caused bronchoconstriction which was antagonized by methysergide, but not modified by pretreating the animals with reserpine.3. In anaesthetized cats, 5,6-dihydroxytryptamine had, in general, a hypotensive effect which was reversed by hexamethonium.4. 5,6-Dihydroxytryptamine also caused aggregation of isolated rabbit and human platelets and inhibited the platelet aggregation induced by 5-hydroxytryptamine (5-HT) plus adrenaline.5. The pattern of action of 5,6-hydroxytryptamine and 5,6-diacetoxytryptamine was qualitatively the same as that of 5-HT, but the potency of the compounds decreased in the order 5-HT, 5,6-dihydroxytryptamine, 5,6-diacetoxytryptamine both in vitro and in vivo.6. It is concluded that 5,6-dihydroxytryptamine and its diacetyl derivative stimulate postsynaptic 5-HT receptors, but that their effect is weaker than that of 5-HT.

Effects of mazindol, a non-phenylethylamine anorexigenic agent, on biogenic amine levels and turnover rate.

1 Mazindol is a new anorexigenic agent which possesses a different chemical structure from that of phenylethylamines, but shows a pharmacological profile similar to that of (+)-amphetamine. 2 Mazindol neither altered whole brain monoamine levels (noradrenaline (NA), dopamine, 5-hydroxytryptamine (5-HT)) nor changed NA levels in the hypothalamus or dopamine levels in the caudate nucleus. 3 Mazindol enhanced dopamine turnover rate in the caudate nucleus, as shown by the increased rate of dopamine decline after blockade of catecholamine synthesis by alpha-methyl-p-tyrosine and decreased the conversion index of (3H)-tyrosine into brain NA. 4 Mazindol administration did not modify pargyline-induced decline of 5-hydroxyindoleacetic acid suggesting that 5-HT turnover is not altered by this drug.

Muscimol antagonism of morphine analgesia in rats.

1 Muscimol, a gamma-aminobutyric acid (GABA) receptor agonist, when injected intraventricularly antagonizes the antinociceptive effect of morphine given either subcutaneously or intraventricularly. The antagonistic effect of muscimol on morphine analgesia appears to be linearly related. 2 This finding provides support for the view that a GABA-ergic system is involved in morphine analgesia.

Modification of the antinociceptive effect of morphine by centrally administered diazepam and midazolam.

1 Intracerebroventricular administration of diazepam or midazolam decreased the antinociceptive effect of morphine in rats as measured by the "tail flick" method. 2 Midazolam, injected into the periaqueductal grey matter (PAG) antagonized the analgesic effect of morphine. The action of midazolam was partially reversed by bicuculline. 3 These findings support the view that the effect of benzodiazephines on morphine antinociception may be mediated through gamma-aminobutyric acid receptors.

Effects of lisuride on body temperature of rats and rabbits: relation to microsomal biotransformation and dopaminergic receptor stimulation.

In rats, lisuride, either administered systemically or intracerebroventricularly induced a dose-related hypothermia. This effect was selectively antagonized by blockade of DA receptors in the CNS but not by inhibition of catecholamine synthesis or blockade of serotoninergic receptors. Also a blocker of "peripheral" DA receptors failed to antagonize the hypothermic effect of lisuride in rats. Induction of rat liver microsomal drug-metabolizing enzymes by phenobarbital counteracted lisuride-induced hypothermia. In rabbits lisuride induced a hyperthermic response which was sensitive to both pimozide and metergoline pretreatment. These findings indicate that stimulation of brain DA receptors involved in thermoregulation is responsible for the changes in body temperature indiced by lisuride in rats and rabbits and that these effects are caused by the drug itself and do not require previous biotransformation into an active metabolite.

Central nervous system neuropeptides after peripheral nerve deafferentation.

Beta-endorphin, Met-enkephalin, substance P, somatostatin and dynorphin concentrations were evaluated in right and left brain areas, and in cervical, thoracic, and lumbosacral spinal cord of rats that underwent section of either the right, the left, both sciatic nerves, the right brachial plexus, the saphenous, or the sural nerve. With all the surgeries, beta-endorphin concentrations decreased significantly in all brain areas with the exception of the striatum where they did not change. By contrast Met-enkephalin increased in all brain areas and in the spinal cord tracts interested by the lesions. The other peptides were always unaffected. The changes in the concentrations of the neuropeptides were observed starting twenty-four hours after surgery and lasted for at least four months. We did not find a lateralization in the brain peptide concentrations of either sham operated or unilaterally deafferentated rats. Moreover, the treatment with serotoninergic agents normalized the concentrations of beta-endorphin, suggesting a role of the serotoninergic system in the decrease of the peptide that follows the lesion of peripheral nerves.

Structural and biochemical alterations in the dorsal horn of the spinal cord caused by peripheral nerve lesions.

Substance P and Met-enkephalin were detected by radioimmunoassay and immunocytochemistry in the rat lumbar spinal cord. The sciatic nerve was lesioned by resecting a piece and the proximal stump was either ligated, for limiting neurite outgrowth, or intraperitoneally sutured, allowing the formation of a large neuroma. Ten days post lesioning both peptide levels dropped approximately 50% and the punctate immunoreactivity decreased in the dorsal horn. Lesioning both sciatic nerves did not accelerate nor increase the extent of peptide loss compared to monolateral lesions. Immunocytochemistry showed that after bilateral lesioning also the punctate immunoreactivity in the dorsal horn decreased less drastically. However, FRAP staining of the dorsal horn decreased according to the lesion paradigms, mono- and bilaterally with the same intensity. Therefore nerve lesions trigger the process, but the peptidergic loss seems intraspinally regulated. In addition, both kinds of abnormal neurite outgrowth similarly altered peptide levels and distribution in the spinal cord. Our data suggest that pain states related to peripheral nerve lesions may be due to opiate peptide loss rather than to neuroma.