George A. Matwyshyn

Dizocilpine (MK-801) Blocks Tolerance to the Analgesic but Not to the Hyperthermic Effect of Morphine in the Rat

The effect of dizocilpine (MK-801), an N-methyl-D-aspartate receptor antagonist, on the development of tolerance to the analgesic and hyperthermic effects of morphine was determined in the rat. Tolerance to morphine in male Sprague-Dawley rats was induced by implanting subcutaneously 6 morphine pellets during a 7-day period. Two schedules of intraperitoneal injections of MK-801 were used. In one, the drug was injected once a day, and in the other it was injected twice a day. The doses of MK-801 were 0.03, 0.1 and 0.3 mg/kg. In the treatment once a day, MK-801 blocked the development of tolerance to the analgesic effect of morphine, but there was no dose-dependent effect. In the treatment twice a day, MK-801 produced a dose-dependent inhibition of tolerance to the analgesic effect of morphine. Higher doses of MK-801 produced high mortality. MK-801 given once a day or twice a day failed to affect the tolerance to the hyperthermic effect of morphine. In both schedules of MK-801 treatment, the highest dose of MK-801 resulted in high mortality. It is concluded that MK-801 is selective in blocking the tolerance to the analgesic effect of morphine in the rat.

Comparative effects of NG-monomethyl-d-arginine and MK-801 on the abstinence syndrome in morphine-dependent mice

The effects of NG-monomethyl-L-arginine (L-NMMA), an inhibitor of nitric oxide (NO) synthase and MK-801, an NMDA receptor antagonist on abrupt and naltrexone-precipitated abstinence symptoms were determined in male Swiss-Webster mice rendered dependent on morphine by subcutaneous implantation of a pellet containing 75 mg of morphine base for 3 days. Mice which served as controls were implanted with placebo pellets. Six hours after pellet removal, mice were injected intraperitoneally with either the vehicle or MK-801 (0.03, 0.1 and 0.3 mg/kg). Thirty minutes later the animals were injected with naltrexone subcutaneously (50 micrograms/kg) and the intensity of abstinence symptoms were determined. Of the three doses of MK-801 used, only 0.1 mg/kg dose inhibited the jumping behavior precipitated by naltrexone in morphine-dependent mice. Whereas the lower dose (0.03 mg/kg) of MK-801 increased, the higher doses of MK-801 (0.1 and 0.3 mg/kg) displayed a decrease in the formation of fecal boli. Administration of MK-801 did not affect the body weight loss observed during abrupt withdrawal (induced by removal of the pellets) in morphine-dependent mice. MK-801 at 0.1 mg/kg dose further decreased the body temperature during abrupt withdrawal in morphine-dependent mice. Other two doses of MK-801 (0.03 and 0.3 mg/kg) did not modify the hypothermia observed during abrupt morphine withdrawal. On the other hand, L-NMMA (0.02 to 4.0 mg/kg) injected intraperitoneally 15 min prior to the naltrexone administration blocked the stereotyped jumping response in a dose-dependent manner. Higher doses of L-NMMA 2.0 and 4.0 mg/kg also decreased the number of fecal boli formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Potentiation of morphine analgesia by BQ123, an endothelin antagonist.

Several neurotransmitter mechanisms have been proposed to play a role in the actions of morphine. The present study is the first to provide evidence that central endothelin (ET) mechanisms are involved in the modulation of pharmacological actions of morphine. The effect of intracerebroventricular (i.c.v.) administration of endothelin-A (ET(A)) antagonist, BQ123, on morphine-induced analgesia, hyperthermia, and catalepsy was determined in the rat. Morphine produced a significant increase in tail-flick latency as compared to control group. Pretreatment with BQ123 significantly potentiated the effect and duration of morphine (2 and 8 mg/kg, s.c.)-induced analgesia as compared to vehicle-pretreated control rats. The hyperthermic effect of morphine was not only significantly greater in BQ123-pretreated rats but also lasted for more than 6 h. ET antagonist, BQ123, did not affect the pharmacological effect of morphine on cataleptic behavior. These studies demonstrate that BQ123, a specific ET(A) receptor antagonist, significantly potentiated morphine-induced analgesia and hyperthermia in rats without affecting morphine-induced cataleptic behavior. [(3)H]-Naloxone binding was carried out to determine the possibility of BQ123 acting on opiate receptors. It was found that morphine could displace [(3)H]-naloxone but BQ123 did not affect [(3)H]-naloxone binding even at 1,000 nM concentration. Therefore, it can be concluded that BQ123 does not act on opioid receptors. This is the first report suggesting that an ET(A) antagonist, BQ123, significantly potentiates the analgesic effect of morphine, possibly through a nonopioid mechanism.

Interactions of thyrotropin releasing hormone, its metabolites and analogues with endogenous and exogenous opiates

The interactions of thyrotropin releasing hormone, its metabolites and synthetic analogues with acute and chronic effects of endogenous and exogenous opiates have been described. The endogenous and exogenous opiates are represented by beta-endorphin and morphine, respectively. The pharmacological effects of opiates include analgesia, temperature effects, respiratory depression, catalepsy, locomotor activity, opiate receptor binding, tolerance, and physical dependence. Thyrotropin releasing hormone and related compounds appear to (a) antagonize hypothermia, respiratory depression, locomotor depression and catalepsy but not the analgesia induced by opiates, (b) inhibit the development of tolerance to the analgesic effect but not to the hypothermic effect of opiates, (c) inhibit the development of physical dependence on opiates as evidenced by the inhibition of development of certain withdrawal symptoms, and (d) suppress the abstinence syndrome in opiate dependent rodents. Thyrotropin releasing hormone does not interact with the opiate receptors in the brain. Potential therapeutic applications of thyrotropin releasing hormone and its synthetic analogues in counteracting some of the undesirable effects of opiates are discussed.

Opioid peptides in pituitary gland, brain regions and peripheral tissues of spontaneously hypertensive and Wistar-Kyoto normotensive rats.

The concentrations of beta-endorphin (beta-END), dynorphin (DYN) and methionine-enkephalin (MEK) in pituitary, brain regions, heart, kidney and adrenal of 8 week old male spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) normotensive rats were determined by radioimmunoassay and compared. The brain regions examined were hypothalamus, striatum, pons + medulla, midbrain and cortex. The concentration of beta-END in pituitary of SHR rats was 49% higher than those of WKY rats. The concentration of beta-END in the striatum of SHR rats was 71% lower as compared to WKY rats. The concentration of beta-END in the heart, adrenals and kidney of SHR rats was significantly lower (92, 48 and 57%, respectively), than those of WKY rat tissues. The concentration of DYN in pituitary, striatum and heart were lower by 38, 55 and 46%, respectively, in SHR compared to WKY rats, but in hypothalamus it was greater (33%) than in WKY rats. The concentration of DYN in other brain areas and in kidney and adrenal did not differ. The tissues of SHR and WKY rats which showed significant difference in the concentration of MEK were pituitary, pons + medulla, cerebral cortex and adrenals. The concentration of MEK was greater in SHR rats with pons + medulla, cortex and adrenals showing 33, 40, 268% higher levels, respectively, over the WKY rat tissues. However, the concentration of MEK in pituitary of SHR rats was 40% lower than that of WKY rats. These studies suggest that the endogenous opioid peptides of both central and peripheral tissues may be important in the regulation of blood pressure in SHR rats.

Endothelin receptor antagonists restore morphine analgesia in morphine tolerant rats

Several neurotransmitter mechanisms have been proposed to play a role in the development of morphine tolerance. The present study provides evidence for the first time that endothelin (ET) antagonists can restore morphine analgesia in morphine tolerant rats. Tolerance to morphine was induced by subcutaneous implantation of six morphine pellets during a 7-day period. The degree of tolerance to morphine was measured by determining analgesic response (tail-flick latency) and hyperthermic response to morphine sulfate (8 mg/kg, subcutaneously (s.c.)) in placebo and morphine pellet implanted rats. The maximal tail-flick latency in morphine pellet-vehicle treated rats (7.54 s) was significantly lower (P<0.05) when compared to placebo pellet-vehicle treated rats (10s), indicating that tolerance developed to the analgesic effect of morphine. In separate sets of experiments, ET antagonists, BQ123 (10 microg, intracerebroventricularly (i.c.v.)) and BMS182874 (50 microg, i.c.v.) were administered in placebo and morphine tolerant rats. BQ123 was injected twice daily for 7 days and once on day 8. BMS182874 was administered only on day 8. Morphine (8 mg/kg, s.c.) was administered 30min after BQ123 or BMS182874 administration. It was found that both BQ123 and BMS182874 potentiated morphine analgesia in placebo and morphine tolerant rats. BQ123 potentiated tail-flick latency by 30.0% in placebo tolerant rats and 94.5% in morphine tolerant rats compared to respective controls. BMS182874 potentiated tail-flick latency by 30.2% in placebo tolerant rats and 66.7% in morphine tolerant rats. Morphine-induced hyperthermic effect was also potentiated by BQ123 and BMS182874. The duration of analgesic action was also prolonged by BQ123 and BMS182874. The effect of BMS182874 was less as compared to BQ123. BQ123 and BMS182874 are selective ET(A) receptor antagonists. Therefore, it is concluded that ET(A) receptor antagonists restore morphine analgesia in morphine tolerant rats.

Regulation of gene expression of various phase I and phase II drug-metabolizing enzymes by tamoxifen in rat liver.

The objective of the present investigation was to evaluate the effect of tamoxifen (TAM) on the gene expression of different phase I and phase II drug-metabolizing enzymes. Groups of male and female F344/NCr rats were administered either corn oil or TAM (2.8 to 45 mg/kg body wt x 14 days) dissolved in corn oil by gavage. An additional group of rats received a diet supplemented with phenobarbital (PB, 500 ppm). Northern blot analyses of total liver RNA were conducted using [32P]-labeled cDNA or oligonucleotide probes coding for different sulfotransferase (ST); UDP-glucuronosyltransferase (UGT), glutathione S-transferase (GST), epoxide hydrolase (EPH) or cytochrome P450 (CYP) mRNA transcripts. In male rats, TAM increased the levels of STel, STa and STpl mRNAs, whereas PB increased only the STel mRNA. In female rats, there was no expression of STel and STHA mRNA in either control or TAM-treated animals. TAM and PB increased UGTBe/p mRNAs in all rats, whereas UGTml mRNA was elevated only in PB-treated animals. EPH mRNA was elevated markedly in all rats treated with TAM and PB, whereas GSTya/ye mRNA was highly increased by PB, but only marginally increased by TAM. Finally, TAM increased CYP3A1 mRNA, and slightly increased CYP2B1 mRNA, whereas PB highly elevated mRNAs for both of these CYP genes. In conclusion, treatments of rats with TAM increased the mRNA levels of many phase I and phase II drug-metabolizing enzymes, and this pleiotypic response to TAM seems to be different from other prototype inducers such as PB or dioxin (TCDD).

Morphine tolerance does not develop in mice treated with endothelin-A receptor antagonists

Long-term use of morphine leads to development of antinociceptive tolerance. We provide evidence that central endothelin (ET) mechanisms are involved in development of morphine tolerance. In the present study, we investigated the effect of ET(A) receptor antagonists, BQ123 and BMS182874, on morphine antinociception and tolerance in mice. Mechanism of interaction of ET(A) receptor antagonists with morphine was investigated. BQ123 (3 microg, i.c.v.) and BMS182874 (50 microg, i.c.v.) significantly enhanced antinociceptive effect of morphine (P < 0.05), through an opioid-mediated effect. Treatment with a single dose of BQ123 (3 microg, i.c.v.) reversed tolerance to morphine antinociception in morphine-tolerant mice. BQ123 or BMS182874 did not affect naloxone binding in the brain. Therefore, ET(A) receptor antagonists did not bind directly to opioid receptors. [35S]GTPgammaS binding was stimulated by morphine and ET-1 in non-tolerant mice. Morphine- and ET-1-induced GTP stimulation was significantly lower (P < 0.05) in morphine-tolerant group (33% and 42%, respectively) compared to control group. BQ123 and BMS182874 did not activate binding in non-tolerant mice. BQ123 and BMS182874 significantly increased G protein activation in morphine-tolerant mice (96% and 86%, respectively; P < 0.05). These results provide evidence that uncoupling of G protein occurs in morphine-tolerant mice, and ET(A) antagonists promote coupling of G protein to its receptors, thereby restoring antinociceptive effect. These findings indicate that ET(A) receptor antagonists potentiate morphine antinociception and reverse antinociceptive tolerance in mice, through their ability to couple G proteins to opioid receptors.

Effects of prolyl-leucyl-glycinamide and cyclo(leucyl-glycine) on morphine-induced antinociception and brain μ, δ and κ opiate receptors

Abstract The effects of prolyl-leucyl-glycinamide and cyclo (leucyl-glycine) on morphine-induced antinociception in mice and on in vitro binding of 3 H-ligands for opiate receptor subtypes (μ, δ and κ) the mouse brain homogenate were determined. Subcutaneous administration of either of the above peptides (1, 2, and 4 mg/kg) 10 min prior to the injection of morphine did not affect morphine-induced antinociception as evidenced by the identical ED 50 values of morphine in vehicle and peptide treated groups. The binding of 3 H-dihydromorphine and 3 H-naloxone ( μ receptors), 3 HDAla 2 DLeu 5 -enkephalin (δ receptors), and 3 H-ethylketocyclazocine (κ receptors) to opiate receptors in the mouse brain homogenate was also unaffected by both the peptides over a large concentration range. It is concluded that these peptides do not interact with brain opiate receptors.

Multiple opiate receptors and pharmacological response to morphine in rats maintained on diets differing in protein concentration

1. The influence of diets differing in protein concentration on the characteristics of mu, delta, and kappa-opiate receptors and on the analgesic and hyperthermic responses to morphine was examined in rats. Three groups of male Sprague-Dawley rats were maintained for four weeks on isocaloric diet containing either 4, 20 or 50% protein. 2. The animals maintained on 4% protein diet weighed 92 +/- 2% of the initial weight at the end of the fourth week, whereas animals maintained on 20% and 50% protein diet weighed 222 +/- 2% and 221 +/- 2%, respectively. The average food intake per 100 g body weight on day 1 of the study in 4, 20 and 50% protein diet group was 5.0 +/- 2.2 g, 10.4 +/- 1.4 g, and 10.0 +/- 1.2 g, respectively. This difference in food intake was not observed during rest of the period of the study. 3. Water intake was higher for the animals maintained on diet containing 50% protein as compared to the other two groups. 4. The analgesic and the hyperthermic response to morphine varied in direct relation to the concentration of protein in the diet. The concentration of morphine in the brain and plasma of animals maintained on the three diets following challenge dose of morphine did not differ. Similarly the Bmax and Kd values for the binding of [3H]naltrexone, [3H]D-Ser2-Thr6-leucine enkephalin and [3H]ethylketocyclazocine to brain membranes prepared from rats kept on 4, 20 and 50% protein concentration did not differ. 5. It is concluded that the diet differing in protein concentration can alter the responses to morphine, and that such altered effects can not be accounted for by the changes in the distribution of morphine in brain and plasma or to the changes in the characteristics of the mu, delta and kappa opiate receptors in the brain.