Gyongyi Horvath

Endomorphin-1 and endomorphin-2: Pharmacology of the selective endogenous μ-opioid receptor agonists

The recently discovered endogenous opioid peptides, endomorphins-1 and -2, appear to have properties consistent with neurotransmitter/neuromodulator actions in mammals. This review surveys the information gained so far from studies of different aspects of the endomorphins. Thus, the endomorphins have been found unequally in the brain; they are stored in neurons and axon terminals, with a heterogeneous distribution; they are released from synaptosomes by depolarization; they are enzymatically converted by endopeptidases; and they interact specifically and with high affinity with mu-opioid receptors. The most outstanding effect of the endomorphins is their antinociceptive action. This depends on both central and peripheral neurons. Additionally, the endomorphins cause vasodilatation by stimulating nitric oxide release from the endothelium. Their roles in different central and peripheral functions, however, have not been fully clarified yet. From a therapeutic perspective, therefore, they may be conceived at present as potent antinociceptive and vasodilator agents.

The Effects of Ketamine and Its Enantiomers on the Morphine- or Dexmedetomidine-induced Antinociception after Intrathecal Administration in Rats

Background The spinal administration of some N-methyl-d-aspartate receptor antagonists results in antinociception and potentiates the effects of opioids and &agr;2-adrenoceptor agonists, but ketamine and its enantiomers have not been examined. The present study investigated the interactions of racemic ketamine, R (−)-ketamine and S (+)-ketamine with morphine and with dexmedetomidine. Methods Intrathecal catheters were implanted into male Wistar rats. Three days later, the acute nociceptive sensitivity was assessed using the tail-flick test. Analgesic latencies were converted to the percentage maximum possible effect. The dose that yielded 50% of the maximum possible effect (ED50) and dose–response and time–course curves were determined for the ketamines (30–300 &mgr;g), morphine (0.1–3.0 &mgr;g), dexmedetomidine (0.3–10.0 &mgr;g), and mixtures of two doses of ketamines (30 or 100 &mgr;g) with different doses of morphine or dexmedetomidine for fixed-dose analysis. Results Neither racemic ketamine nor its enantiomers alone had a significant effect on the tail-flick test, with the exception of the highest dose of racemic ketamine, which caused motor impairment. Morphine and dexmedetomidine each produced dose-dependent antinociception, with ED50 of 1.7 &mgr;g (95% confidence interval: 1.04–2.32) and 4.85 &mgr;g (3.96–5.79), respectively. A low dose (30 &mgr;g) of racemic ketamine or its enantiomers did not influence the ED50 of morphine significantly. Coadministration of 100 &mgr;g racemic ketamine or S (+)-ketamine, but not R (−)-ketamine, significantly enhanced and prolonged the antinociceptive effect of morphine. Both doses of racemic ketamine or its isomers significantly decreased the ED50 value for dexmedetomidine, although the higher dose of racemic or S (+)-ketamine had the highest potency. One-hundred micrograms of racemic ketamine or S (+)-ketamine also prolonged the effects of dexmedetomidine. Conclusions These data indicate that racemic ketamine and S (+)-ketamine, but not R (−)-ketamine, exhibit similar effectiveness in potentiating the antinociceptive effects of both morphine and dexmedetomidine.

The role of TRPV1 receptors in the antinociceptive effect of anandamide at spinal level

&NA; While it is well known that the endogenous cannabinoid receptor ligand anandamide also activates the transient receptor potential vanilloid1 (TRPV1) receptors, there has been no in vivo study indicating the role of the TRPV1 receptors in the antinociceptive effect of anandamide at spinal level. The goal of this study was to determine the effect of inhibition of TRPV1 receptors by capsazepine on the antinociceptive potency of anandamide after intrathecal administration. Anandamide alone (1, 30 or 100 μg) dose‐dependently decreased carrageenan‐induced thermal hyperalgesia, however, the highest dose caused temporary excitation and vocalization, suggesting the pain‐inducing potential of anandamide. Capsazepine (10 or 20 μg) by itself did not change the pain sensitivity markedly, but the lower dose increased it, and the higher dose decreased the antinociceptive effect of 30 μg anandamide. Furthermore, both doses of capsazepine decreased the efficacy of the largest dose of anandamide. These results show that TRPV1 receptor activation plays a substantial role in the antinociceptive effects of anandamide at spinal level. The effect of the inhibition on TRPV1 receptors depended on the dose applied. We presume that coactivation of the cannabinoid and TRPV1 receptors by anandamide provides elevated antinociception through the release of antinociceptive endogenous ligands at spinal level.

The synergistic antinociceptive interactions of endomorphin-1 with dexmedetomidine and/or S(+)-ketamine in rats.

Spinal administration of the endogenous &mgr;-opioid agonist peptide, endomorphin-1, results in antinociception in rodents, but there are few data about its interaction with other antinociceptive drugs. We investigated the antinociceptive interactions at the spinal level of endomorphin-1 with the N-methyl-d-aspartate antagonist S(+)-ketamine, the &agr;2-adrenoceptor agonist dexmedetomidine, or both in awake rats. Nociception was assessed by the tail-flick test. Dose-response curves were determined for endomorphin-1 (0.6–50 &mgr;g), for dexmedetomidine (0.1–10 &mgr;g), for mixtures of S(+)-ketamine (30 or 100 &mgr;g) with endomorphin-1 (2–18 &mgr;g) or of endomorphin-1 with dexmedetomidine in a fixed ratio (4:1), and for the triple combination of the three drugs after intrathecal administration. Endomorphin-1 and dexmedetomidine both produced dose-dependent antinociception. The coadministration of 100 &mgr;g S(+)-ketamine significantly enhanced the antinociceptive effect of 6 &mgr;g endomorphin-1. Isobolographic analysis of the combinations of endomorphin-1 and dexmedetomidine revealed a synergistic interaction between these drugs. The 80% effective dose for the triple combination was significantly less than that for either binary combination. These data indicate that S(+)-ketamine and dexmedetomidine, acting via different receptors, produce synergistic antinociceptive interaction with endomorphin-1 at the spinal level. Furthermore, the triple combination of an opioid agonist, an &agr;2-adrenoceptor agonist, and an N-methyl-d-aspartate receptor antagonist shows potent antinociceptive activity.

Antinociceptive effect of the S(+)-enantiomer of ketamine on carrageenan hyperalgesia after intrathecal administration in rats

Ketamine exerts antinociceptive effects in many pain tests.We investigated the antinociceptive effect of intrathecally administered racemic ketamine and its S(+)- and R(-)-enantiomer on carrageenan-induced thermal hyperalgesia with a paw withdrawal test and acute pain (hot plate and tailflick) tests. Rats were prepared with a chronic lumbar intrathecal catheter to receive either saline or ketamine enantiomers in cumulative doses. None of the ketamines (10, 50, or 100 [micro sign]g) had any effect on the withdrawal latency of the contralateral, noninjected paw. In the injected paw, intrathecal saline did not alter carrageenan-induced thermal hyperalgesia, whereas intrathecally applied S(+)-, R(-)-, and racemic ketamine decreased thermal hyperalgesia. However, compared with saline, racemic ketamine had a higher efficacy than S(+)-ketamine, whereas R(-)-ketamine did not achieve statistical significance. Neither S(+)- nor R(-)-ketamine had a significant effect in the tailflick test (10, 100, or 500 [micro sign]g). In the hot plate test, only the largest dose of ketamine (500 [micro sign]g) caused a non-stereospecific, significant increase in hot plate latency; this dose caused supraspinal effects as well. The results demonstrate that the behavioral hyperalgesia associated with carrageenan-induced hindpaw inflammation in rats is attenuated by the intrathecal administration of racemic and S(+)-ketamine, but not R(-)-ketamine, which only displayed an insignificant trend toward a dose-response relationship. This finding warrants further studies to investigate a possible clinical advantage of preservative-free S(+)-ketamine over the currently used preservative containing racemic mixture. Implications: In rats, intrathecal S(+)-ketamine was effective for treating inflammatory pain. Although racemic ketamine has a greater efficacy, S(+)-ketamine is available as a preservative-free drug and might be of clinical interest for future neuraxial administration in different pain states. (Anesth Analg 1998;86:561-5)

Anti-inflammatory effects of phosphatidylcholine in neutrophil leukocyte-dependent acute arthritis in rats

We investigated the effects of exogenous phosphatidylcholine (PC) and non-steroidal diclofenac supplementation on polymorphonuclear cell influx in carrageenan-induced arthritis in rats. The microcirculatory consequences were evaluated by a novel method developed for direct intravital microscopic observation of the synovial membrane. Arthritis was induced by injection of a mixture of 2% lambda-carrageenan and 4% kaolin into the knee joints and the animals were treated orally with PC (150 mg/kg twice daily), sodium diclofenac (0.5mg/kg twice daily) or saline vehicle. Intravital videomicroscopy was used to investigate the leukocyte-endothelial interactions directly in the synovial membrane at 6h after the challenge. The inflammation-induced thermal and mechanical secondary hyperalgesic reactions were assessed at 24h, and the knee volume changes at 48h after the insult. The development of arthritis was accompanied by a significant increase in the number of adherent leukocytes in the synovial postcapillary venules, but this increase was reduced significantly (by approximately 40%) by PC, and slightly (by 22%) by diclofenac treatment. The perivascular infiltration of the neutrophil leukocytes and the intercellular adhesion molecule-1 (ICAM-1) expressions were reduced only by PC treatment. The significant decrease (45%) in the thermal nociceptive latency, the 3-fold increase in the mechanical touch sensitivity and the knee cross-sectional area (which was increased by 35% by the arthritis induction) were significantly ameliorated by both treatments. The present study demonstrated the anti-inflammatory effects of PC in experimental arthritis. The therapeutic potential may be linked to the reduction of neutrophil leukocyte-mediated microcirculatory inflammatory reactions.

Effect of intrathecal agmatine on inflammation-induced thermal hyperalgesia in rats

Agmatine, an endogenous ligand, interacts both with the alpha2-adrenoceptors and with the imidazoline binding sites. The effect of intrathecally administered agmatine on carrageenan-induced thermal hyperalgesia was investigated by means of a paw-withdrawal test in rats. The effect of agmatine on morphine-induced anti-hyperalgesia was also studied. Intrathecal agmatine in doses larger than 250 microg caused a decrease in the pain threshold, with vocalization and agitation lasting for several hours in all animals. Agmatine alone at 1-100 microg did not give rise to any change in the thermal withdrawal threshold in the contralateral non-inflamed paw. Agmatine pretreatment was found to dose-dependently attenuate the thermal hyperalgesia induced by intraplantar carrageenan. The effect of 100 microg agmatine was completely lost by 60 min, whereas the effect of 50 microg was of similar magnitude but exhibited a longer duration. Agmatine posttreatment had a slighter effect. Agmatine pretreatment (100 microg) together with 1 microg morphine (subeffective dose) has significantly higher anti-hyperalgesic effect then the individual compounds by themselves. These are the first data demonstrating the behavioral and anti-hyperalgesic effects of intrathecal agmatine. The results reveal important interactions between intrathecal agmatine and opioids in thermal hyperalgesia.

Uncoupling protein 2 (UCP2) lowers alcohol sensitivity and pain threshold

Abuse of ethanol is a major risk factor in medicine, in part because of its widespread effect on the activity of the central nervous system, including behavior, pain, and temperature sensation. Uncoupling protein 2 (UCP2) is a mitochondrial protonophore that regulates cellular energy homeostasis. Its expression in mitochondria of axons and axon terminals of basal forebrain areas suggests that UCP2 may be involved in the regulation of complex neuronal responses to ethanol. We employed a paradigm in which acute exposure to ethanol induces tolerance and altered pain and temperature sensation. In UCP2 overexpressing mice, sensitivity to ethanol was decreased compared to that of wild-type animals, while UCP2 knockouts had increased ethanol sensitivity. In addition, UCP2 expression was inversely correlated with the impairment of pain and temperature sensation induced by ethanol. Taken together, these results indicate that UCP2, a mitochondrial uncoupling protein previously associated with peripheral energy expenditure, is involved in the mediation of acute ethanol exposure on the central nervous system. Enhancement of UCP2 activation after acute alcohol consumption might decrease the time of recovery from intoxication, whereas UCP2 inhibition might decrease the tolerance to ethanol.

High-dose Remifentanil Does Not Impair Cerebrovascular Carbon Dioxide Reactivity in Healthy Male Volunteers

Background Cerebrovascular carbon dioxide reactivity during high-dose remifentanil infusion was investigated in volunteers by measurement of regional cerebral blood flow (rCBF) and mean CBF velocity (CBFv). Methods Ten healthy male volunteers with a laryngeal mask for artificial ventilation received remifentanil at an infusion rate of 2 and 4 &mgr;g · kg−1 · min−1 under normocapnia, hypocapnia, and hypercapnia. Stable xenon-enhanced computed tomography and transcranial Doppler ultrasonography of the left middle cerebral artery were used to assess rCBF and mean CBFv, respectively. If required, blood pressure was maintained within baseline values with intravenous phenylephrine to avoid confounding effects of altered hemodynamics. Results Hemodynamic parameters were maintained constant over time. Remifentanil infusion at 2 and 4 &mgr;g · kg−1 · min−1 significantly decreased rCBF and mean CBFv. Both rCBF and mean CBFv increased as the arterial carbon dioxide tension increased from hypocapnia to hypercapnia, indicating that cerebrovascular reactivity remained intact. The average slopes of rCBF reactivity were 0.56 ± 0.27 and 0.49 ± 0.28 ml · 100 g−1 · min−1 · mmHg−1 for 2 and 4 &mgr;g·kg−1·min−1 remifentanil, respectively (relative change in percent/mmHg: 1.9 ± 0.8 and 1.6 ± 0.5, respectively). The average slopes for mean CBFv reactivity were 1.61 ± 0.95 and 1.54 ± 0.83 cm · s−1 · mmHg−1 for 2 and 4 &mgr;g · kg−1 · min−1 remifentanil, respectively (relative change in percent/mmHg: 1.86 ± 0.59 and 1.79 ± 0.59, respectively). Preanesthesia and postanesthesia values of rCBF and mean CBFv did not differ. Conclusion High-dose remifentanil decreases rCBF and mean CBFv without impairing cerebrovascular carbon dioxide reactivity. This, together with its known short duration of action, makes remifentanil a useful agent in the intensive care unit when sedation that can be titrated rapidly is required.

Selective disturbance of pain sensitivity after social isolation.

The effects of social isolation or NMDA-receptor antagonists on pain sensitivity have repeatedly been described. However, the mechanisms underlying the alterations of pain perception in these models still remain a matter of debate. Thus, we aimed to determine the long-lasting effects of subchronic ketamine treatment and social isolation on the C- and Adelta-fiber-mediated nociception. Wistar rats after weaning (21-23 days old) were either housed individually or grouped for 21 days. The animals were treated daily for 14 days with either ketamine (30 mg/kg) or saline. On the 21st day, tail-flick latency was determined at 48 degrees C (C-fiber activation) and 52 degrees C (affects mainly Adelta-fibers), and rats were rehoused. Tail-flick test was repeated 2 and 4 weeks later. On the 5th week, carrageenan-induced heat hyperalgesia was determined on paw-withdrawal test before and after morphine treatment (1, 2 or 3 mg/kg). Regarding tail-flick latencies at 48 degrees C, juvenile isolation, but not ketamine resulted in a significantly enhanced pain threshold (p<0.001) throughout the investigation period, while the changes at 52 degrees C were not significant. In addition, both isolation and ketamine treatments enhanced the antihyperalgesic effect of 2 mg/kg morphine. In summary, juvenile isolation exerts a long-lasting effect on acute heat pain sensitivity, disturbing primarily the C-fiber-linked pain pathways, suggesting a selective disruption in the parallel sensory pathways. Since both social isolation and NMDA treatment are well-known animal models of schizophrenia, our results showed that juvenile isolation but not ketamine administration can simulate hypoalgesia associated with this disease.