Search

The effects of i.c.v. administration of several aminoglycoside antibiotics on naloxone-precipitated morphine withdrawal symptoms were evaluated in mice acutely dependent on morphine. Neomycin (10-40 micrograms/mouse), gentamicin (40-160 micrograms/mouse) and kanamycin (80-320 micrograms/mouse) produced a dose-dependent reduction of the number of precipitated jumps, forepaw tremors and head shakes. The order of potency of the aminoglycoside antibiotics on all withdrawal symptoms was neomycingentamicinkanamycin, which is the same order that these drugs show as N-type calcium channel blockers. The capacity of several drugs that decrease neuronal calcium availability (such as lanthanum and L-type calcium channel blockers) to antagonize opiate withdrawal is well known. In the light of these findings, our results suggest that the mechanism of aminoglycoside-induced inhibition of morphine abstinence may be related to the capacity of these antibiotics to block N-type calcium channels, and to decrease neuronal calcium availability.

1. The analgesic effects of diltiazem and verapamil, both per se and together with morphine, were studied using subcutaneous (s.c.) and intracerebroventricular (i.c.v.) administrations, in the hot-plate test in mice. 2. The i.c.v. injection of verapamil (15-120 micrograms/mouse) and diltiazem (60-120 micrograms/mouse) induced dose-dependent analgesic effects. 3. The i.c.v. administration of verapamil (30-120 micrograms/mouse) and diltiazem (60-120 micrograms/mouse) significantly enhanced, in a dose-dependent way, the analgesic effects of morphine and produced a parallel displacement to the left of the morphine log dose-response line. 4. When these calcium channel blockers were administered subcutaneously at doses of 40 and 80 mg/kg, they exerted no analgesic actions, but dose-dependently potentiated the analgesic effects of morphine, producing a parallel shift to the left of the morphine log dose-response line. 5. These results suggest that inhibition of calcium entry through calcium channels induced by verapamil and diltiazem may play a role in analgesia development.

The effect of the non-competitive NMDA receptor antagonist dizocilpine (MK-801) on conditioned place preference induced by morphine was studied in mice. As expected, morphine (1-8 mg/kg, i.p.) elicited a significant preference for the drug-paired compartment. Pretreatment of mice with (+)-dizocilpine (0.1 and 0.2 mg/kg, i.p.), the more active dizocilpine enantiomer, dose-dependently reversed the conditioned place preference produced by morphine (4 mg/kg, i.p.), whereas (-)-dizocilpine (0.2 mg/kg, i.p.) did not modify morphine-induced effects. In contrast, both enantiomers of dizocilpine (at a dose of 0.2 mg/kg, i.p.) elicited a conditioned place preference. These data suggest that (1) NMDA receptors play a role in morphine-induced place preference, and (2) dizocilpine-reinforcing properties in the place preference paradigm do not seem to be dependent on NMDA receptor blockade.

The influence of the ATP-sensitive K+(KATP) channel opener cromakalim on the antinociception induced by agonists of several receptors coupled to pertussis toxin-sensitive G proteins, clonidine (alpha2 adrenoceptor), baclofen (gamma-aminobutyric acid(B) receptor), morphine (mu opioid receptor) and U50,488H (kappa opioid receptor), was evaluated with a tail-flick test in mice. The subcutaneous administration of clonidine (0.12-2 mg/kg), morphine (0.5-16 mg/kg), baclofen (2-16 mg/kg) and U50,488H (2-16 mg/kg) induced a dose-dependent antinociceptive effect. Cromakalim (8-64 microgram/mouse intracerebroventricularly [i.c.v.]) did not change tail-flick latency in control animals but produced a dose-dependent enhancement of the antinociception induced by clonidine and morphine, and shifted their dose-response curves to the left. These effects of cromakalim were antagonized dose dependently by the K(ATP) channel blocker gliquidone (0.1-8 microgram/mouse i.c.v.). On the other hand, cromakalim (16-64 microgram/mouse i.c.v.) did not significantly enhance the antinociception induced by baclofen and U50,488H and did not shift their dose-response curves. These results suggest that opening of the K(ATP) channels plays an important role in the antinociception mediated by alpha(2) adrenoceptors and mu opioid receptors, but not in that induced by gamma-aminobutyric acid(B) and kappa opioid receptors.

The modulation by K+ channel-acting drugs of the antinociceptive effect of several 5-HT1A receptor agonists was examined with the hot plate test in mice. All the 5-HT1A receptor agonists tested induced dose-dependent antinociception, the order of potency being (+/-)-8-hydroxy-2-(di-n-propyl-amino)tetralin (8-OH-DPAT) > buspirone > or = lesopitron > or = tandospirone. The blockers of ATP-sensitive K+ channels (KATP) gliquidone and glipizide (1-4 and 16-64 micrograms/mouse i.c.v., respectively) reduced the antinociceptive effect of 8-OH-DPAT, whereas cromakalim (32-64 micrograms/mouse i.c.v.), an opener of KATP channels, enhanced the effect. In contrast, 4-aminopyridine (25-250 ng/mouse i.c.v.) and tetraethylammonium (10-20 micrograms/mouse i.c.v.), which antagonize several non-ATP-dependent K+ conductances, were inactive. The same results were found with other agonists of 5-HT1A receptors (lesopitron, buspirone and tandospirone): gliquidone inhibited whereas cromakalim increased their antinociceptive effects. None of the K+ channel-acting drugs modified the binding of [3H]8-OH-DPAT to hippocampal membranes, whereas all the 5-HT1A receptor agonists displaced the ligand. These results suggest that ATP-sensitive K+ conductances are involved in the antinociception induced by agonists of 5-HT1A receptors.

We studied the effects of two different ATP-sensitive K+ channel openers on naloxone-precipitated withdrawal in morphine-dependent mice. The i.c.v. administration of cromakalim and diazoxide (both at 5-40 micrograms/mouse) dose-dependently inhibited several signs of morphine withdrawal (number of jumps and episodes of forepaw tremors, and body weight loss). At present it is impossible to specify the exact mechanism(s) involved in this effect. However, considering that morphine opens K+ channels in neurons, it is tempting to suggest that K+ channel openers can mimic the effects of morphine on neuronal K+ currents, and as a consequence can act as substitutes for this drug during morphine withdrawal.

The effect of gliquidone, an ATP-dependent K+ (KATP) channel blocker, on morphine-induced hypermotility in mice was studied. Morphine (5-40 mg/kg s.c.) dose dependently increased ambulatory activity. Gliquidone (10 micrograms/mouse i.c.v.) induced a parallel displacement to the right of the morphine dose-response curve. Moreover, gliquidone (10 and 40 micrograms/mouse i.c.v.) produced a dose-dependent antagonism of morphine (20 mg/kg s.c.)-induced hypermotility. These results suggest that KATP channels are involved in morphine-induced hypermotility. The present data, together with those of previous studies showing antagonism by KATP channel blockers of morphine-induced antinociception and hyperthermia, further indicate that the opening of KATP channels plays an important role in the mechanism of action of morphine.

The effects of four ATP-dependent K+ channel blockers (hypoglycemic sulfonylureas) against morphine- and U50488H-induced antinociception were evaluated using the tail flick test in mice. None of the sulfonylureas tested significantly modified tail flick latency in control animals. However, i.c.v. pretreatment with gliquidone (0.4-1.6 micrograms/mouse), glipizide (2.5-10 micrograms/mouse), glibenclamide (10-40 micrograms/mouse) or tolbutamide (20-80 micrograms/mouse) dose dependently antagonized morphine-induced antinociception approximately equieffectively, the only difference being in potency: gliquidoneglipizideglibenclamidetolbutamide. This effect of sulfonylureas was very specific, since none antagonized the antinociception elicited by U50488H even at doses twice as great as the dose that induced maximum antagonism of morphine antinociception. Because morphine, but not U50488H, opens K+ channels in neurons and because the order of potency of the different sulfonylureas for blocking ATP-dependent K+ channels in neurons and for antagonizing morphine antinociception is the same, we suggest that morphine antinociception is mediated by the opening of ATP-dependent K+ channels.

The effects of several calcium antagonists (verapamil, nicardipine and two diltiazem isomers, d-cis and l-cis diltiazem) alone and associated to non-depolarizing (pancuronium) and depolarizing (succinylcholine) neuromuscular blockers, were evaluated on sciatic nerve-tibialis anterior muscle preparations from cats in vivo. The calcium antagonists used (at 0.1 and 0.5 mg/kg iv) did not modify the height of muscular twitches elicited indirectly. However, these agents potentiated in a dose-dependent way the neuromuscular blockade induced by iv pancuronium (2-40 micrograms/kg) and succinylcholine (6-200 micrograms/kg). The order of potency in increasing the effects of pancuronium was nicardipine much greater than d-cis diltiazem greater than or equal to verapamil, whereas the order of potency in enhancing succinylcholine effects was d-cis diltiazem greater than or equal to verapamil much greater than nicardipine. The effects of diltiazem were stereoselective, thus the potentiation induced by d-cis diltiazem was significantly greater in all cases than that induced by l-cis diltiazem, which suggests that calcium channel blockade plays a role in these interactions. However, other mechanisms such as calcium antagonists-induced nicotinic receptor desensitization may also be involved.

The possible analgesic effects of i.c.v. administration of several aminoglycoside antibiotics were evaluated in mice using hot plate and tail flick tests. Neomycin (10-80 micrograms/mouse), gentamicin (40-160 micrograms/mouse) and kanamycin (80-320 micrograms/mouse) produced dose-dependent increases in the latencies to forepaw licking and jumping in hot plate test. These drugs also produced dose-dependent increases in the percentage of animals showing analgesia in tail flick test. The order of potency of these aminoglycoside antibiotics in both tests was neomycin greater than gentamicin greater than kanamycin, which is exactly the same order that these drugs show as N-type calcium channel blockers. Bearing in mind this fact and the well known analgesic activity of several drugs which decrease neuronal calcium availability, we suggest that the mechanism of aminoglycoside-induced antinociception may be related to the capacity of these antibiotics to block N-type calcium channels and decrease neuronal calcium availability.

We compared the effects of nicardipine on indirectly and directly elicited twitches in phrenic-hemidiaphragm preparations from rats, and the effects of calcium and Bay K 8644 on nicardipine-induced neuromuscular blockade. Nicardipine (5-250 microM) decreased both directly and indirectly elicited muscular contractions in a concentration-dependent way. The inhibitory effect was greater when the preparation was indirectly stimulated. Nicardipine antagonism of indirectly elicited contractions was reversed in an apparently competitive way by calcium (1-4 mM). In contrast, the calcium channel agonist Bay K 8644 (0.1 and 1 microM) had no effects per se, nor did it reverse neuromuscular blockade induced by nicardipine. Interestingly, Bay K 8644, at 10 microM, decreased the height of indirectly elicited muscular contractions. These results suggest that the calcium antagonist nicardipine affects neuromuscular transmission to a greater degree than muscular contraction, this effect being antagonized by calcium. In contrast, the calcium agonist Bay K 8644 was unable to produce any effect as stimulator of calcium channels at the neuromuscular level, indicating that this drug may exhibit tissue specificity.

We showed that glibenclamide, a blocker of ATP-dependent potassium channels in the CNS, antagonizes morphine-induced analgesia in mice

The effects of two calcium channel blockers (verapamil and flunarizine) were evaluated on the naloxone-precipitated syndrome in morphine-dependent rats. The withdrawal signs in saline-treated rats were mainly diarrhea, body weight loss, jumping and ptosis. On i.p. administration, verapamil and flunarizine prevented diarrhea and body weight loss but not jumping. Verapamil also reduced the incidence of ptosis at the highest dose tested (40 mg/kg). Administered i.c.v., 160 μg verapamil reduced the body weight loss and the number of jumps without modifying diarrhea or ptosis. The results show that calcium channel blockers inhibit morphine abstinence syndrome manifestations through both peripheral and central mechanisms.

The effects of calcium channel blockers and stimulants on naloxone-precipitated morphine withdrawal in morphine-dependent ileum were evaluated in vitro. Both verapamil and diltiazem (0.01–1 μM) inhibited the naloxone-induced morphine withdrawal contractures in a concentration-dependent way. The effect of diltiazem was stereospecific. On the other hand, the calcium channel stimulant, Bay k 8644 (0.01 μM), significantly increased the naloxone-induced contractures of morphine-dependent ileum. These results a suggest a role for calcium channels in morphine withdrawal in vitro

The in vitro effects of flunarizine on indirectly- and directly-elicited contractions in rat phrenic-hemidiaphragm preparations were studied. The interactions of flunarizine with non-depolarizing and depolarizing neuromuscular blocking drugs (pancuronium and suxamethonium) and with an aminoglycoside antibiotic (neomycin) were also evaluated. Flunarizine induced a slowly developing concentration-dependent reduction of indirectly-elicited diaphragm twitch height, but only slightly reduced directly-elicited contractions. Flunarizine 1 and 5 mumol.l-1 produced a concentration-dependent enhancement of pancuronium-induced neuromuscular blockade, whereas suxamethonium blockade was significantly increased by flunarizine 5 mumols.l.-1 only. Moreover, both flunarizine 1 and 5 mumols.l-1 also increased the neuromuscular blockade induced by neomycin. In conclusion, flunarizine induced neuromuscular blockade and enhanced the effects of several neuromuscular blocking agents to varying degrees in vitro.

The effects were studied of several calcium channel blocking agents on muscle twitch, and possible interactions between these drugs and pancuronium and suxamethonium, using a rat phrenic-hemidiaphragm preparation. Nicardipine, verapamil and diltiazem each caused a concentration-related depression of muscle response. Nicardipine had the most, and diltiazem the least, potent effect. Verapamil and diltiazem 5 and 10 mumol litre-1 caused a concentration-dependent enhancement of suxamethonium-induced neuromuscular blockade, but increased the effect of pancuronium only at 10 mumol litre-1. Nicardipine 10 mumol litre-1 significantly enhanced pancuronium-induced neuromuscular blockade, but not that produced by suxamethonium.

The effects of several calcium channel blockers (nifedipine, verapamil and diltiazem) on rat phrenic-hemidiaphragm preparations were studied. The calcium channel blockers were used either alone or associated with two aminoglycoside antibiotics, neomycin and streptomycin. All drugs investigated produced a concentration-dependent decrease in indirectly elicited diaphragmatic contractions. The order of potency was: verapamil greater than neomycin congruent to nifedipine greater than diltiazem greater than streptomycin. Moreover, neomycin-induced neuromuscular blockade was significantly increased by nifedipine (1 and 10 microM), verapamil (1 and 10 microM) and diltiazem (10 microM), whereas the streptomycin-induced neuromuscular blockade was increased only by nifedipine (1 and 10 microM) and verapamil (10 microM).

The effects of two enantiomers of diltiazem (l-cis and d-cis) on naloxone-precipitated morphine abstinence after acute morphine dependence were evaluated in vivo in mice and in vitro in isolated pieces of rat terminal ileum. d-cis-diltiazem (10-40 mg/kg) produced a dose-dependent inhibition of the jumping and body weight loss induced by naloxone in acutely morphine dependent mice. By contrast, l-cis-diltiazem 40 mg/kg did not significantly inhibit jumping and produced a much lower inhibition of body weight loss than the same dose of d-cis-diltiazem. In addition, d-cis-diltiazem (0.01-1 microM) produced a concentration-dependent inhibition of naloxone-induced contracture in morphine dependent ilea, whereas l-cis-diltiazem, even at 1 microM, did not inhibit this contracture. These results suggest that calcium channels may play a similar role in naloxone-precipitated morphine abstinence in vivo and in vitro.

The possibility that calcium channel blockers might produce antinociception and increase morphine analgesia was examined using the acetic acid writhing test in mice. Subcutaneous injections of diltiazem, verapamil, nicardipine, flunarizine and cinnarizine produced a dose-dependent antinociception. This activity of diltiazem was stereospecific; d-cis-diltiazem was more potent than 1-cis-diltiazem. All the calcium channel blockers studied increased morphine analgesia and displaced to the left the morphine dose-response curve. This effect of diltiazem was also stereospecific. These results suggest that calcium channel blockers can induce analgesia and increase morphine analgesia, possibly through a decrease in cellular calcium availability.