RB101-mediated protection of endogenous opioids: potential therapeutic utility?

The endogenous opioids met- and leu-enkephalin are inactivated by peptidases preventing the activation of opioid receptors. Inhibition of enkephalin-degrading enzymes increases endogenous enkephalin levels and stimulates robust behavioral effects. RB101, an inhibitor of enkephalin-degrading enzymes, produces antinociceptive, antidepressant, and anxiolytic effects in rodents, without typical opioid-related negative side effects. Although enkephalins are not selective endogenous ligands, RB101 induces these behaviors through receptor-selective activity. The antinociceptive effects of RB101 are produced through either the mu-opioid receptor alone or through activation of both mu- and delta-opioid receptors; the antidepressant-like and anxiolytic effects of RB101 are mediated only through the delta-opioid receptor. Although little is known about the effects of RB101 on other physiologically and behaviorally relevant peptides, these findings suggest that RB101 and other inhibitors of enkephalin-degrading enzymes may have potential as novel therapeutic compounds for the treatment of pain, depression, and anxiety.

Through central arginine vasopressin, not oxytocin and endogenous opiate peptides, glutamate sodium induces hypothalamic paraventricular nucleus enhancing acupuncture analgesia in the rat.

Our previous study proved that the hypothalamic paraventricular nucleus (PVH) plays an important role in acupuncture analgesia. The neuropeptides involving in the PVH regulation of acupuncture analgesia was investigated in the rat. The changes of pain threshold, which was induced by electrical acupuncture of "Zusanli" points (St. 36), were measured as acupuncture analgesia. Microinjection of l-glutamate sodium into the PVH, which only excites the PVH neurons, could dose-dependently enhance the acupuncture analgesia, but microinjection of l-glutamate sodium into the area nearby the PVH did not alter acupuncture analgesia. Removing pituitary did not influence this effect of l-glutamate sodium. Microinjection of l-glutamate sodium into the PVH only increased the arginine vasopressin (AVP), not oxytocin (OXT), leucine enkephaline (L-Ek), beta-endorphine (beta-Ep) and dynorphinA(1-13) (DynA(1-13)) concentrations in the PVH perfuse liquid using radioimmunoassay. Intraventricular injection of anti-arginine vasopressin serum (AAVPS) could completely reverse the effect of microinjection of l-glutamate sodium into the PVH enhancing acupuncture analgesia. Intraventricular injection of naloxone, one opiate peptide antagonist, partly attenuated this effect of l-glutamate sodium, and intraventricular of anti-oxytocin serum (AOXTS) did not change this effect of l-glutamate sodium. The results suggested that l-glutamate sodium induces the PVH enhancing acupuncture analgesia only through AVP, not OXT and endogenous opiate peptides in central nervous system.

A possible role for an enkephalinergic system in the internal defense mechanism of Biomphalaria alexandrina exposed to Schistosoma mansoni.

The present study documents that in Bioniphalaria alexandrina coordinated responses to Schistosoma mansoni infection are modulated by receptor-mediated opioid signals. Rather comprehensive tests in susceptible and resistant snails have demonstrated: I- the presence of an endogenaus opioids in the snail hemolymph (in particular, Leu-enkephalin-like material). II- in vitro treatment of snail hemocytes with synthetic Leu-enkephalin analogue (DADLE) resulted in the modulation of cellular adherence, and phagocytic activity. III- the addition of Naloxone, either alone or in combination whith DADLE, generally reduced hemocyte activity indicating opioid-receptor-mediated mechanism. V- the presence of DADLE or Naloxone modulated the level of IL-2-, TNF-gamma- and FNF-alpha-like molecules in S. mansoni resistant and susceptible snails. Specifically, DADLE and DADLE in combination with Naloxone generally were found to be capable of modulating resistant snail hemocytes at concentrations of 10(-6) and 10(-8) M. Similar actions after incubation with the same concentrations were not detected in the susceptible snails. These observations demonstrate the existence of a complete opioid system in B. alexandrina, associated with susceptibility and resistance to S. mansoni infection, the results suggest the role of such opioid system in molecular signaling within the host and in host-parasite interactions.

Toluene alters brainstem enkephalinergic system in rats.

Acute exposure to high doses of toluene can generate respiratory depression. However, neurotoxic mechanism of its action in the brainstem is not completely clear. In this work, acute, but not subchronic, exposure of rats to toluene increased leu-enkephalin immunostaining in several myelencephalic nuclei implicated in cardiorespiratory control. Due to the physiological role of enkephalins in the central regulation of breathing, it is suggested that the enkephalinergic system could play a role in neurotoxic respiratory depression induced by high dose acute toluene exposure.

Pituitary adenylate-cyclase activating polypeptide (PACAP) evokes long-lasting secretion and de novo biosynthesis of bovine adrenal medullary neuropeptides.

Recently, the pituitary adenylate-cyclase activating polypeptide (PACAP) has emerged as a potential noncholinergic neuromodulator of adrenal medullary function. In support of this hypothesis, we documented PACAP's effects on the secretion and biosynthesis of neuropeptides by cultured bovine chromaffin cells. Data presented in this study indicate that PACAP is a potent and efficacious secretagogue of leucine-enkephalin which was coreleased with catecholamines with identical profiles. In comparison to nicotinic activation, however, rates of PACAP-induced secretion were substantially slower but persisted for several hours causing a prolonged increase in the tonic release of both transmitters and peptides. Interestingly, renewal of intracellular pools of neuropeptides was also stimulated by PACAP but not the vasoactive intestinal peptide (VIP). Indeed, the higher incorporation of [35S]-labeled amino acids into atrial and brain natriuretic peptides (ANP, BNP) provided strong evidence that PACAP directly activated de novo biosynthesis. Of particular importance was PACAP's net preferential stimulation of the biosynthesis of BNP, similar to the differential regulation by protein kinase A (PK-A) and protein kinase C (PK-C) activators we have previously the differential regulation by protein kinase A (PK-A) and protein kinase C (PK-C) activators we have previously reported. PACAP-induced secretion and biosynthesis appeared to be mediated by the PACAP-specific type I receptors known to activate adenylate cyclase and phospholipase C. We verified that PACAP did indeed stimulate the production of cyclic AMP and inositol phosphates in our cell system. These findings suggest that the dual signaling properties of type I receptors may be important for PACAP's differential effect on the biosynthesis of natriuretic peptides. We conclude that PACAP might assume important noncholinergic trans-synaptic regulation of the adrenal medulla by releasing and modifying intragranular catecholamine and neuropeptide contents.

[The interaction of the renin-angiotensin and enkephalinergic systems of the brain and hypophysis in normal rats and in experimental pathology of the hypothalamo-hypophyseal-adrenal complex]. / Vzaimodeistvie renin-angiotenzinovoi i énkefalinergicheskoi sistem mozga i gipofiza krys v norme i pri éksperimental'noi patologii gipotalamo-gipofizarno-nadpochechnikovogo kompleksa.

Administration of angiotensin-converting enzyme inhibitor captopril inhibited the activity of renin-angiotensin system (RAS), increased leukin-enkephalin level in the hypothalamus and adenohypophysis. The data obtained suggest that the enzyme is an important link in the mechanism of interaction between the renin-angiotensin and enkephalinergic systems in rats.

[The activity of the enkephalinergic systems of the brain and hypophysis in experimental hypocorticism]. / Aktyvnist' enkefalinerhichnoï systemy mozku ta hipofiza za umov eksperymental'noho hipokortytsyzmu.

Changes in activity of basic components of enkephalinergic system, leu-enkephalin contents, activity of enkephalin-hydrolysing enzymes (enkephalinases A and B, enkephalin aminopeptidases) and 3H-leu-enkephalin specific binding to opioid receptor in rat anterior and mediobasal hypothalamus, striatum, medulla oblongata and adenohypophysis have been analysed on experimental models of hypocorticoidism. No changes in brain and pituitary body leu-enkephalin contents following unilateral hypocorticoidism. No changes in brain and pituitary body leu-enkephalin contents following unilateral adrenalectomy were shown. Bilateral adrenalectomy resulted in two-phase character of neuropeptide level: a decrease of leu-enkephalin contents in hypothalamus, striatum and adenohypophysis on the 7th day and its increase to the normal level on the 10th day after the operation were revealed. A decrease of leu-enkephalin contents in rat brain on the 7th day following adrenalectomy occurred simultaneously with a decrease in enkephalin aminopeptidase activity and specific binding of labeled leu-enkephalin testifying to strengthening of enkephalin release form neurosecretory granules of brain structures following adrenalectomy. Important changes in leu-enkephalin contents, reception and inactive processes on the level of adenohypophysis and on the level of hypothalamus, striatum and medulla oblongata were detected by cortisol and ACTH administration in adrenalectomised animals.

[The effect of the aminopeptidase inhibitor bestatin on the enkephalin level in the rat brain]. / Vliianie ingibitora aminopeptidaz bestatina na uroven' énkefalinov v mozge krys.

Met- and leu-enkephalin contents in midbrain (including hypothalamus) and striatum of rats were determined by radioimmunoassay after bestatin (racemate) injection (200 g, i.c.v.). It was found that bestatin administration influenced the midbrain met-enkephalin content, values and directions of the changes observed being dependent upon the time after the injection. The data obtained confirm the participation of aminopeptidase in enkephalin inactivation and present evidence for the possibility of regional variations of enkephalin catabolism pathways in the brain.