The effects of p-chlorophenylalanine on morphine analgesia,tolerance and dependence development in two strains of rats

The effects of p-chlorophenylalanine (p-CPA; 100 mg/kg/day for 3 days) on morphine analgesia and the development of tolerance and physical dependence were investigated in Sprague-Dawley (SD) and Fisher (F) strains of albino rats. Using a modified flinch-jump method to detect changes in reactivity to electric footshock, F strain rats were more reactive to the footshock than SD rats, but showed less relative increase in threshold (analgesia) than SD rats following various doses of morphine. Pretreatment with p-CPA attenuated significantly morphine analgesia in SD, but not F rats. In animals implanted subcutaneously with morphine pellets, p-CPA appeared to delay the development of tolerance to morphine in both strains of rats. Hyperalgesia and loss of body weight resulted from administration of naloxone to pellet-implanted rats and p-CPA pretreatment lessened these withdrawal effects significantly in SD rats only. These results emphasize the importance of strain differences in the study of morphine analgesia and development of tolerance and dependence. Assuming differences in the function of the serotonergic inhibitory system in the two strains of rats, these data provide general support for the involvement of brain 5-HT mechanisms in modulating, if not mediating the effects of morphine.     

Comparison of tolerance to morphine-induced respiratory and analgesic effects in mice

Morphine is responsible for severe poisonings in chronically treated patients. We hypothesize that toxicity could be related to the development of weaker tolerance for morphine-induced deleterious respiratory effects in comparison to analgesic effects. Our objectives were to compare tolerance to both effects in mice and investigate possible mechanisms for such possible differences. Tolerance to morphine-induced analgesia and respiratory effects was assessed using hot plate response latencies and plethysmography, respectively. Mechanisms of tolerance were investigated using binding studies to mu-opioid receptors (MOR) and adenylate cyclase (AC) activity measurement in homogenates of cell membranes from the periaqueductal gray region (PAG) and brainstem. Morphine (2.5 mg/kg) was responsible for analgesia with significant increase in inspiratory time. Acute tolerance to analgesia (p < 0.01) and effects on respiratory frequency (p < 0.05) was observed in mice pre-treated with 100 mg/kg morphine in comparison to saline. Following repetitive administration (2.5 mg/kg/day during 10 days), we observed a 13-fold increase in the effective dose-50% (ED₅₀) of morphine-induced analgesia in comparison to a 2- or 4-fold increase in the ED₅₀ of its related increase in inspiratory time determined in air and 4% CO₂, respectively. No significant alteration in MOR expression was observed in either PAG or brainstem following repeated morphine administration. However, in PAG, in contrast to brainstem, superactivation of AC was observed in morphine-treated mice in comparison to controls (p < 0.05). In conclusion, tolerance to morphine-induced respiratory effects is much more limited than tolerance to its analgesic effects in repeatedly morphine-treated mice. The difference in morphine-induced AC activation between the brainstem and the PAG contributes to the observed difference in tolerance between both morphine effects.     

Modification of morphine-induced analgesia, tolerance and dependence by bromocriptine

The effect of two doses of bromocriptine, a dopamine agonist, on morphine-induced analgesia, tolerance and dependence was investigated in mice. Bromocriptine at doses of 0.04 and 0.08 mg/kg did not affect the baseline tail flick latency of mice but potentiated the morphine analgesia. Pretreatment of mice with 5 mg/kg of sulpiride, a D-2 antagonist, not only blocked the effect of 0.08 mg/kg of bromocriptine but also antagonized the morphine analgesia. Control animals given daily injections of 10 mg/kg of morphine rapidly developed tolerance to the analgesic effect. A combined treatment of bromocriptine with morphine given daily suppressed the development of tolerance to morphine analgesia. However, development of tolerance to morphine analgesia was not significantly modified in the animals treated daily with bromocriptine (0.08 mg/kg) plus sulpiride (5 mg/kg). Acute dependence was induced by the administration of 100 mg/kg of morphine. The administration of bromocriptine 30 min before naloxone significantly decreased the ED₅₀ value for naloxone for inducing jumping in mice. Coadministration of sulpiride and bromocriptine attenuated the ability of bromocriptine to potentiate the withdrawal syndrome of morphine dependence. The results indicate that bromocriptine potentiates morphine analgesia, suppresses the development of tolerance to morphine analgesia but exacerbates opiate withdrawal signs in morphine-dependent mice. These effects of bromocriptine appear to be mediated via D-2 receptors.     

Bupropion attenuates morphine tolerance and dependence: Possible role of glutamate,norepinephrine, inflammation,and oxidative stress

Background

Morphine – the main pillar of nociceptive pain management – systemic use is associated with development of tolerance and dependence. Tolerance and dependence lay a heavy burden in clinical pain management settings. An added weight to this dilemma is that effective, safe, and tolerable solution to this problem is still beyond reach. Antidepressants were reported as possible alleviators of opioid tolerance and dependence. One of the increasingly used antidepressant in clinical practice is bupropion given its high safety and tolerability profile.

The effects of receptor selective opioid peptides on morphine-induced analgesia

Utilizing the mouse tail-flick assay, four opioid peptides, which have been reported to be selective for either μ- or δ-opioid receptors, were examined for their analgesic potency and for their ability to modify morphine-induced analgesia. [D-Ala²,D-Leu⁵]enkephalin and [D-Ser²,Thr⁶]leucine-enkephalin, putative δ-receptor selective peptides, produced a potent analgesic response and at subanalgesic doses potentiated morphine-induced analgesia. Morphiceptin and [D-Ala²,Pro⁵]enkephalinamide, putative μ-receptor selective peptides, were similarly found to produce analgesia. However, in contrast to the δ-receptor selective peptides, three μ-receptor selective peptides were unable to alter the potency of morphine. Thus, it would appear that the potentiation of morphine analgesia is a unique property of δ-receptor selective peptides.     

Facilitation of morphine-induced tolerance and physical dependence by prolyl-leucyl-glycinamide

The pretreatment of mice with 30 mg/kg morphine s.c. did not alter the analgesic effect of morphine in mice pretreated with saline but decreased the analgesic effect of morphine in mice pretreated with prolyl-leucyl-glycinamide (PLG). Tolerance was evaluated by the effect of PLG on morphine-induced enhancement of naloxone potency which is a measure of the capacity of naloxone to antagonize morphine-induced analgesia and is postulated to be an indicator of tolerance development. The naloxone potency of PLG-treated mice was 2-fold greater than that of control mice. PLG did not alter the whole brain levels of morphine, nor did it alter the naloxone potency in mice which were not pretreated with 30 mg/kg morphine. In mice treated with 100 mg/kg morphine or implanted with 50 mg morphine pellets for 24 or 72 h, the amount of naloxone required to induce jumping was not altered by PLG. However, PLG treatment did increase the hypothermia and body weight loss seen after naloxone-induced withdrawal. Administration of PLG to morphine-dependent mice 1 h prior to naloxone did not modify the resultant hypothermia or body weight loss. These results indicate that PLG facilitated the development of morphine tolerance and dependence.     

Time-dependent disruptive effects of apomorphine and alpha-methyl-p-tyrosine on development of morphine tolerance

In rats the development of one-trial tolerance to the analgesic action of i. p. administered morphine is disrupted by the postadministration of apomorphine (30 mg/kg at 5 min but not at 3 h) or alpha-methyl-p-tyrosine (200 mg/kg at 5 min and 3 h but not at 12 h). Diethyldithiocarbamate or propranol has no disruptive effect. It is suggested that development of tolerance to the analgesic effects of morphine is mediated by sequential time-dependent biochemical processes.     

Interaction between ACTH fragments, brain opiate receptors and morphine-induced analgesia.

The present study confirms that N-terminal fragments of ACTH have an affinity for rat brain opiate receptors in vitro. Such peptides, devoid of corticotrophic activity, were found to inhibit morphine-induced analgesia if they also possessed affinity for opiate receptors in vitro. The structure-activity relationship for these two parameters is comparable to that observed for the same peptides on the induction of excessive grooming.     

Modification of morphine analgesia and tolerance by flumazenil in male and female rats

This study assessed the effect of the central benzodiazepine receptor antagonist, 8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]benzodiazepine-3-carboxylic acid ethyl ester (flumazenil), on morphine-induced analgesia, locomotor effects, and development of tolerance in rats. The thermally evoked pain (tail flick) response was determined after acute and chronic intraperitoneal (i.p.) administration of morphine and flumazenil, alone and in combination. In acute studies, flumazenil induced weak analgesia unrelated to dose and sex, whereas morphine-induced analgesia was dependent on both dose and sex (male>female). Flumazenil dose-dependently enhanced the analgesic effect of morphine in female but not in male rats. Isobolographic analysis suggested synergism between flumazenil and morphine in female rats, but antagonism in male rats. Flumazenil-induced locomotor changes (alone and with morphine) were related to sex but not dose. Chronic coadministration of flumazenil with morphine enhanced analgesia and attenuated tolerance development in female rats. The findings suggest a possible role for flumazenil as an adjunct with opioids in acute and chronic pain therapy.     

Comparative effects of prolyl-leucyl-glycinamide and naloxone on morphine-induced inhibition of gastrointestinal transit

The effects of prolyl-leucyl-glycinamide (MIF) and naloxone on the gastrointestinal transit in mice were investigated using the charcoal meal test. MIF administered intraperitoneally (IP) (1-10 mg/kg) or intracerebroventricularly (ICV) (10 micrograms/mouse) had no effect on the transit. Administration of morphine by subcutaneous (SC) route significantly inhibited the gastrointestinal transit. The morphine-induced inhibition of the transit was not affected by MIF whether given by IP or ICV route. Administration of the opiate antagonist naloxone (1 mg/kg, IP or 10 micrograms/mouse, ICV) had no effect on the gastrointestinal transit, but it significantly antagonized the inhibition produced by morphine. Some earlier studies have indicated narcotic antagonistic effect of MIF. However, in the present study, evidence for such an action of MIF was not obtained. It is suggested that MIF does not appear to have narcotic antagonistic activity and further supports an earlier study from this laboratory that MIF may not interact with opiate receptors.     

The role of vasopressin on the effect of U-50,488 to block the development of morphine tolerance and physical dependence

U-50,488, a selective κ-opioid receptor agonist, has been reported to inhibit the development of antinociceptive tolerance to morphine in mice, rats and guinea pigs, but the mechanism involved in this action remains unknown. Since U-50,488 has been reported to supress the plasma vasopressin level, we investigated the role of vasopressin with U-50,488 in the male Sprague Dawley rat in this study. Animals (230–270 g) were chronically treated with morphine (10 mg/kg, i.p.) twice a day for 6 days in order to induce tolerance to antinociceptive effect measured by tail-flick test. Withdrawl symptoms were precipitated by naloxone (10 mg/kg, i.p.) on day 7. U-50,488 (i.p.) or AVP (i.p. or i.c.v.) or U-50,488 and AVP was (were) coadministered with chronic morphine to investigate their effects on morphine tolerance and dependence. We found that coadministration of 8 mg/kg U-50,488 (i.p.) with morphine almost completely block morphine tolerance and partially block withdrawal symptoms. In contrast, coadministration of AVP (0.3 μg/kg, i.p., or 0.01 μg, i.c.v.) with morphine and U-50,488, the effects of U-50,488 to block morphine tolerance and dependence were reversed. In addition, treatment of AVP antagonist (dPTyr(Me)AVP, 0.5 μg/kg, i.p. or 0.5 μg, i.c.v.) has the similar effect as U-50,488 to block morphine tolerance. In summary, the effect of U-50,488 to block morphine tolerance and dependence may relate to its inhibitory effect on AVP release. Received: 20 February 1996 / Accepted: 14 October 1996     

Morphine tolerance and dependence in noradrenaline neurones of the rat cerebral cortex

Summary By subcutaneous implantation of 2 or 13 morphine pellets (75 mg morphine/pellet), rats were made tolerant to, and dependent on narcotic analgesics. Occipital cortex slices from dependent animals and placebo-implanted controls were incubated with (-)-³H-noradrenaline and subsequently superfused with physiological salt solution. The accumulation of ³H-noradrenaline was not changed by pretreatment with 2, but was slightly decreased by pretreatment with 13 morphine pellets. The overflow of tritium evoked by electrical field stimulation was higher in slices from morphine-implanted rats than in those from placebo controls. Morphine and levorphanol, added in vitro, inhibited the stimulation-induced overflow of tritium at, similar concentrations and to a similar degree in slices from morphine-and placebo-pretreated animals. —It is concluded that, during chronic treatment with morphine, an adaptation takes place in the brain to compensate for the acute effect of narcotic analgesics, i.e. inhibition of the release of noradrenaline by nerve impulses. The chain of events from the drug-receptor interaction to the depression of the release process can be excluded as substrate of this adaptation. During with-drawal, the compensatory changes provoke an enhanced increase of extracellular noradrenaline during nerve impulses.     

Influence of hemicholinium (HC-3) on morphine analgesia, tolerance, physical dependence and on brain acetylcholine

Administration of HC-3 intracerebrally at a dose which reduced brain acetylcholine (ACh) without any change in choline (Ch) levels antagonized morphine antinociception slightly as measured by inhibition of the tail-flick response, in both naive mice and in mice rendered tolerant to and dependent on morphine by pellet implantation. However, the development of tolerance to morphine and of dependence on morphine were not affected by HC-3. Although naloxone precipitated withdrawal jumping was enhanced irrespective of whether the HC-3 was administered before or after the dependence on morphine had developed, body weight loss during abrupt withdrawal was unaffected by HC-3. The results indicate that although some of the acute and withdrawal effects of morphine may be associated either directly or indirectly with acetylcholine, these actions do not appear to be the primary process responsible for initiating the development of tolerance and dependence.     

Influence of prazosin and clonidine on morphine analgesia,tolerance and withdrawal in mice

Rapid development of tolerance and dependence limits the usefulness of morphine in long-term treatment. We examined the effects of clonidine (₂-adrenoceptor agonist) and prazosin (₁-adrenoceptor antagonist) on morphine analgesia, tolerance and withdrawal. Morphine tolerance was induced using a 3-day cumulative twice-daily dosing regimen with s.c. doses up to 120 mg/kg. Tolerance was assessed on day 4, as loss of the antinociceptive effect of a test dose of morphine (5 mg/kg). After 10 h, morphine withdrawal was precipitated with naloxone (1 mg/kg). Prazosin had no analgesic effect alone but dose-dependently potentiated morphine analgesia in morphine-naive mice. Another ₁-adrenoceptor antagonist, corynanthine, had similar effects. Prazosin also increased the analgesic potency of the morphine test dose in morphine-tolerant mice. Naloxone-precipitated vertical jumping was not affected, but weight loss was reduced by prazosin. Acutely administered clonidine potentiated morphine analgesia and alleviated opioid withdrawal signs, as expected. We conclude that in addition to the already established involvement of ₂-adrenoceptors in opioid actions, also ₁-adrenoceptors have significant modulatory role in opioid analgesia and withdrawal.     

A genetic analysis of morphine-induced running and analgesia in the mouse

A genetic analysis of morphine-induced analgesia and activity was conducted in mice belonging to the strains BALB/cJ, C57BL/6J, DBA/2J and to their F1 and backcross progenies. The results support previous findings showing that a negative correlation is evident between these two behavioral measures and support that their mode of inheritance is characterized by dominance or partial dominance. The biometric analysis conducted on the parental, F1 hybrid and backcross populations indicates very clearly that the effects of morphine are genetically determined.     

The influence of learning on morphine analgesia and tolerance development in rats tested on the hot plate

The influence of learning on the development of tolerance to the analgesic effect of morphine in rats was examined employing the hot plate procedure. A tested-reinforced (Tr) group and its yoked-control, a tested-non-reinforced (Tnr) group, received identical exposure to the testing procedure; the Tr group was reinforced daily for its behavior on the heated plate whereas the Tnr group was reinforced only on the last day of the experiment. Paired statistical comparisons between these two groups on the last day of the experiment revealed that: 1. premorphine control reaction times on the heated plate were significantly lower in Tr than in Tnr animals; and 2. post-morphine increases in reaction time did not differ between Tr and Tnr animals. It was concluded that whereas some learning does occur in this testing procedure, learning does not influence the behavioral tolerance to morphine which develops in this analgesiometric method. An hypothesis which accommodates this behavioral tolerance and a mechanistic scheme is offered.     

Discriminative stimulus properties of fentanyl and morphine: tolerance and dependence.

Using a food-reinforced two-lever operant procedure, rats were trained to discriminate 0.04 mg/kg fentanyl from saline. At different time intervals after the establishment of discriminative responding, stimulus generalization experiments were performed with equivalent dose ranges of fentanyl (0.0025 to 0.02 mg/kg) and morphine (2.5 to 20 mg/kg). It was found that the ED50 values of both compounds for generalization with the narcotic discriminative stimulus complex, did not change over a 4-month period. The subjects used in this study demonstrated marked tolerance to narcotic analgesia, but none of them showed signs reminescent of narcotic dependence. It is concluded that the discriminative stimulus properties of narcotic analgesics are not subject to tolerance.     

Differential effects of opiate agonists-antagonists on morphine-induced hyperexcitability and analgesia in mice

The effects of two opiate agonists-antagonists, butorphanol (4.0 and 8.0 mg/kg) and buprenorphine (0.1 and 1.0 mg/kg), were assessed on locomotor activity and analgesia in DBA/2 and C57BL/6 mice. Diferrent behavioral effects were evident in these strains, which might becharacterized by different reactions to the effects of opiates and by differences in endorphin distributions and opiate receptor populations. In particular, buprenorphine acted as an agonist-antagonist to morphine in both strains while a dissociation of buturphanol effects was evident, depending on the strain considered. The clinical implications of these findings are discussed.     

Blockade of orexin receptor 1 attenuates the development of morphine tolerance and physical dependence in rats

The goals of this study were to evaluate the effects of pretreatment by orexin receptor-1 antagonist on the development of morphine tolerance and physical dependence in rat. Animals were rendered dependent on morphine by subcutaneous (SC) injection of morphine sulfate (10 mg/kg) at set intervals of 12 h for 10 days. Just before the morphine administration, the animals received SB-334867, a selective orexin receptor 1 (OXR1) antagonist. To assess morphine tolerance, the antinociceptive responses of morphine were measured using the warm-water tail immersion test before and after its administration. On day 11, naloxone was injected 2 h after morphine administration and the physical dependence evaluated by quantifying/scoring naloxone-precipitated withdrawal signs for 30 min. The effect of chronic SB-334867 on locomotion was carried out by calculating the number of grid crossings as a measure of locomotor activity. Our findings demonstrated that although morphine-tolerance tended to develop in response to repeated injections of morphine, pre-treatment of OXR1 antagonist prevented this effect, causing a delay in the development of morphine-tolerance. Moreover, co-administration of orexin receptor 1 antagonist with morphine significantly decreased the somatic signs of withdrawal including diarrhea, teeth chattering, jumping, and defecation. Administration of SB-334867 alone or in a chronic co-administration with morphine failed to change locomotor activity. These results suggest that the activation of OXR1 might be involved in the development of morphine tolerance and dependence.     

Antagonism of morphine analgesia by prolyl-leucyl-glycinamide (MIF-1) in humans

Prolyl-leucyl-glycinamide (MIF-1) has been observed to inhibit the analgesic effect of morphine in a series of animal studies. In the present study, the naloxone-like properties of MIF-1 were assessed in human subjects. Eight men received a capsule containing 60 mg of MIF-1 or placebo followed one hour later by a 10 mg intramuscular injection of morphine in a double-blind, crossover design at two visits 4 weeks apart. Experimental pain was induced by the cold pressor test administered 45, 75, 120 and 180 min after the morphine. Each subject recorded severity of pain on a 100 mm line scale every 5 sec during the 120 sec his foot was immersed in the cold water tank and during the 60 seconds immediately following its removal. On a third visit, baseline values were measured in the absence of morphine, MIF-1 or placebo. Analysis of variance revealed that MIF-1 resulted in significantly higher scores (less analgesia) compared with placebo when measured at 45 and 75 min after morphine during the immersion phase and during all four times the subjects were evaluated during the removal phase. The results indicate that MIF-1 can act in humans as an opiate antagonist.