nitroarginine and Pain

These electrophysiological studies, using desensitization and selective antagonists, demonstrate a peripheral role of bradykinin in the generation of pain in the rat. In addition, nitric oxide is shown to play a complex role in peripheral and spinal events in nociception.

Topics: Animals; Arginine; Bradykinin; Formaldehyde; Inflammation; Neurons; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitroarginine; Pain; Rats; Receptors, Bradykinin; Receptors, Neurotransmitter; Spinal Cord

Downstream regulatory element antagonistic modulator (DREAM) is a critical transcriptional repressor for pain modulation. The role of nitric oxide (NO) plays in modulating DREAM pain pathway in the periphery is unclear. Therefore, we investigated the role of the NO in modulation of the expression of DREAM in formalin-induced rat inflammatory pain models. Male Sprague-Dawley rats were randomly distributed into four groups: the normal group, formalin test group, Nω-nitro-L-arginine (l-NNA) group, and morphine group. One hundred microliters of 2.5 % formalin was injected into the plantar surface of the right hindpaw of rats. l-NNA (40 nmol/L) and morphine (40 nmol/L) were injected intrathecally in the hindpaw before formalin injection. The nociceptive behavioral reaction was recorded. After the formalin test, the expression of DREAM mRNA and protein in the spinal cord of the four groups were measured. The nociceptive reaction induced by injection of formalin exhibited two phases. Morphine and l-NNA significantly decreased pain scores of the second phase. The expression of DREAM was significantly increased in the rat spinal cord after formalin-induced pain. Morphine significantly upregulated the expression of DREAM, and the formalin-induced upregulation was significantly attenuated by l-NNA. NO may play an important role in the DREAM pathway modulation of inflammatory pain.

Topics: Animals; Formaldehyde; Inflammation; Injections, Spinal; Kv Channel-Interacting Proteins; Male; Morphine; Nitric Oxide; Nitroarginine; Pain; Pain Measurement; Random Allocation; Rats; Rats, Sprague-Dawley; Repressor Proteins; RNA, Messenger; Spinal Cord

In experiments on female rats it has been shown that the blockade of nitric oxide synthesis by the intraperitoneal administration of NO-synthase non-selective inhibitor nitro-L-arginine at the dose of 10 mg/kg before and after stress (produced by 6, or 12, or 24 h immobilization with painful stimulus) results in the decrease of cardiac ino- and chronotropic functions. In carrying out stress tests (the tests of adrenoreactivity and maximum isometric workload) a considerable decrease in increment of myocardial contractility findings and frequency of heart contractions has been found in animals after 6, 12, 24 h stress under the conditions of nitric oxide synthesis suppression in comparison to stressed animals of control group.

Topics: Animals; Blood Pressure; Epinephrine; Female; Heart; Heart Rate; Immobilization; Injections, Intraperitoneal; Myocardial Contraction; Nitric Oxide; Nitric Oxide Synthase; Nitroarginine; Pain; Periodicity; Rats; Stress, Physiological

The involvement of the L-arginine/nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway in antinociception has been implicated as a molecular mechanism of antinociception produced by several antinociceptive agents, including μ-, κ-, or δ-opioid receptor agonists, nonsteroidal analgesics, cholinergic agonist, and α2C adrenoceptor agonist. In this study, we investigated whether ketamine, a dissociative anesthetic N-methyl-D-aspartate receptor antagonist, was also capable of activating the L-arginine/NO/cGMP pathway and eliciting peripheral antinociception.. The rat paw pressure test was used, with hyperalgesia induced by intraplantar injection of prostaglandin E2. All drugs were locally administered into the right hindpaw of male Wistar rats.. Ketamine (10, 20, 40, 80 μg/paw) elicited a local antinociceptive effect that was antagonized by the nonselective NOS inhibitor L-NOARG (12, 18, and 24 μg/paw) and by the selective neuronal NOS inhibitor L-NPA (12, 18, and 24 μg/paw). In another experiment, we used the inhibitors L-NIO and L-NIL (24 μg/paw) to selectively inhibit endothelial and inducible NOS, respectively. These 2 drugs were ineffective at blocking the effects of the peripheral ketamine injection. In addition, the level of nitrite in the homogenized paw indicated that exogenous ketamine is able to induce NO release. The soluble guanylyl cyclase inhibitor ODQ (25, 50, and 100 μg/paw) blocked the action of ketamine, and the cGMP-phosphodiesterase inhibitor zaprinast (50 μg/paw) enhanced the antinociceptive effects of low-dose ketamine (10 μg/paw).. Our results suggest that ketamine stimulates the L-arginine/NO/cyclic GMP pathway via neuronal NO synthase to induce peripheral antinociceptive effects.

Topics: Analgesia; Anesthetics, Dissociative; Animals; Arginine; Cyclic AMP; Dinoprostone; Drug Synergism; Enzyme Inhibitors; Excitatory Amino Acid Antagonists; Guanylate Cyclase; Hyperalgesia; Ketamine; Male; Nitric Oxide; Nitric Oxide Synthase; Nitroarginine; Oxadiazoles; Pain; Peripheral Nervous System Diseases; Quinoxalines; Rats; Rats, Wistar; Receptors, N-Methyl-D-Aspartate; Signal Transduction

The objective of this study was to extend our previous findings by investigating in greater detail the mechanisms that might be involved in the antinociceptive action of p-methoxyl-diphenyl diselenide, (MeOPhSe)(2), in mice. The pretreatment with nitric oxide precursor, l-arginine (600 mg/kg, intraperitoneal, i.p.), reversed antinociception caused by (MeOPhSe)(2) (10 mg/kg, p.o.) or N(G)-nitro-l-arginine (l-NOARG, 75 mg/kg, i.p.) in the glutamate test. Ondansetron (0.5 mg/kg, i.p., a 5-HT(3) receptor antagonist) and SCH23390 (0.05 mg/kg, i.p.., a D(1) receptor antagonist) blocked the antinociceptive effect caused by (MeOPhSe)(2). Conversely, pindolol (1 mg/kg, i.p., a 5-HT(1A)/(1B) receptor/beta adrenoceptor antagonist), WAY 100635 (0.7 mg/kg, i.p., a selective 5-HT(1A) receptor antagonist), ketanserin (0.3 mg/kg, i.p., a selective 5-HT(2A) receptor antagonist), prazosin (0.15 mg/kg, i.p., an alpha(1)-adrenoreceptor antagonist), yohimbine (1.0 mg/kg, i.p., an alpha(2)-adrenoreceptor antagonist), sulpiride (5 mg/kg, i.p., a D(2) receptor antagonist), naloxone (1 mg/kg, i.p., a non-selective opioid receptor antagonist) and caffeine (3 mg/kg, i.p., a non-selective adenosine receptor antagonist) did not change the antinociceptive effect of (MeOPhSe)(2). (MeOPhSe)(2) significantly inhibited nociception induced by intraplantar (i.pl.) injection of bradykinin (10 nmol/paw) and Des-Arg(9)-bradykinin (10 nmol/paw, a B(1) receptor agonist). (MeOPhSe)(2) significantly inhibited phorbol myristate acetate (PMA, 0.03 mug/paw, a protein kinase C (PKC) activator)-induced licking response. These results indicate that (MeOPhSe)(2) produced antinociception in mice through mechanisms that involve an interaction with nitrergic system, 5-HT(3) and D(1) receptors. The antinociceptive effect is related to (MeOPhSe)(2) ability to interact with kinin B(1) and B(2) receptors and PKC pathway mediated mechanisms.

Topics: Analgesics; Animals; Arginine; Benzazepines; Benzene Derivatives; Bradykinin; Dopamine Antagonists; Enzyme Inhibitors; Glutamic Acid; Magnetic Resonance Spectroscopy; Male; Mice; Neurotransmitter Agents; Nitric Oxide; Nitric Oxide Synthase; Nitroarginine; Organoselenium Compounds; Pain; Receptors, Neurotransmitter; Tetradecanoylphorbol Acetate

The role of central and peripheral neuronal nitric oxide synthase (nNOS) splice variants in the development of inflammatory hyperalgesia was investigated using the formalin test. Supraspinal administration of the NOS inhibitor NOArg lowered both the first and second phase of the formalin response. An oligodeoxynucleotide targeting four nNOS isoforms given supraspinally also reduced the formalin response of both phases. Supraspinal antisense mapping suggested that this effect results from the nNOS-1 splice variant, implying that nNOS-1 is important in mediating formalin pain. At the spinal level, antisense mapping suggested a role of both the nNOS-1 and the nNOS-beta variants in producing formalin pain. Conversely, an antisense selective against nNOS-2 had an opposing effect against the first phase, increasing its intensity. This result, which was similar to prior studies examining opioid actions, implies that endogenous nNOS-2 activity acted to minimize pain perception. Locally in the foot, arginine, the precursor for NO, increased the phase II response at low doses while higher doses reduced the response. This complex biphasic response suggested opposing NOS actions. Local antisense mapping again showed that nNOS-1 is involved in producing phase II of the formalin response while nNOS-2 had an opposite effect similar to that seen spinally. Finally, downregulation of nNOS-1 by antisense prevented tolerance to morphine in both the tail-flick and the formalin test. Together, these observations illustrate the complexity of nNOS in pain perception and the existence of opposing nNOS systems likely due to splice variants of nNOS.

Topics: Analysis of Variance; Animals; Arginine; Drug Tolerance; Formaldehyde; Hyperalgesia; Injections, Spinal; Isoenzymes; Male; Mice; Morphine; Nitric Oxide Synthase Type I; Nitroarginine; Oligodeoxyribonucleotides, Antisense; Pain; Pain Measurement; Pain Threshold

Painful reaction of rats to intraperitoneal injections of L-arginine, Nw-nitro-L-arginine, and agmatine was studied on the model of formalin-induced inflammation. All drugs exhibited a dubious effect on the patterns of nociceptive behavior depending on the phase of painful reaction. The dynamics of nitrate/nitrite content in animal blood and serum indicated the presence of NO-dependent and NO-independent components in the mechanisms of pharmacological effects of these drugs.

Topics: Agmatine; Animals; Arginine; Behavior, Animal; Formaldehyde; Inflammation; Injections, Intraperitoneal; Male; Nitric Oxide; Nitroarginine; Pain; Pain Measurement; Rats; Rats, Wistar; Spectrophotometry

We have previously verified that niga-ichigoside F(1) (NI), a triterpene isolated from Rubus imperialis, exhibits significant and potent antinociceptive action when evaluated in some pharmacological models of pain in mice. This effect was confirmed in other experimental models and also the mechanism of action has been evaluated. The antinociception caused by NI (60 mg kg(-1)) in both phases of the formalin test was significantly attenuated by intraperitoneal injection of mice with haloperidol (a dopaminergic antagonist, 0.20 mg kg(-1)) and L-arginine (precursor of nitric oxide, 600 mg kg(-1)). Regarding the cholinergic system, atropine (a cholinergic antagonist 60 mg kg(-1)) reverted only the second phase. The effect of NI was not affected by treatment of mice with yohimbine (an alpha2-adrenoceptor antagonist, 0.15 mg kg(-1)). The same pharmacological profile was observed for the administration of naloxone (an opioid receptor antagonist, 1 mg kg(-1)). On the other hand, intraperitoneal injection caused dose-related and significant effects against glutamate- and capsaicin-induced pain, respectively. In conclusion, the marked antinociception of NI appears to be related to the dopaminergic, cholinergic, glutamatergic, tachykininergic and oxinitrergic systems, supporting the ethnomedical use of Rubus imperialis (Rosaceae).

Topics: Analgesics; Animals; Apomorphine; Atropine; Capsaicin; Dose-Response Relationship, Drug; Drug Administration Schedule; Female; Glutamic Acid; Haloperidol; Injections, Intraperitoneal; Male; Mice; Molecular Structure; Morphine; Naloxone; Nitroarginine; Pain; Pain Measurement; Plant Extracts; Rosaceae; Saponins; Yohimbine

The modulation by spinal nitric oxide (NO) of descending pathways travelling through the dorsal lateral funiculus (DLF) is a mechanism proposed for the antinociceptive effects of drugs that changes the NO metabolism. In this study we confirm that a surgical incision in the mid-plantar hind paw of rats reduces the threshold to mechanical stimulation with von Frey filaments. The incisional pain was further increased in rats with ipsilateral DLF lesion. Intrathecal L-NOARG (50-300 microg), or SIN-1 (0.1-5.0 microg) reduced, while SIN-1 (10 and 20 microg) intensified the incisional pain in rats with sham or effective lesion of the DLF. Stimulation of the dorsal raphe (DRN) or anterior pretectal (APtN) nuclei with stepwise increased electrical currents (7, 14, 21, 28 and 35 microA r.m.s.) produced a current-related reduction of the incisional pain. These nuclei activate pain inhibitory pathways that descend to the spinal cord mainly through the DLF. Intrathecal SIN-1 (5 microg) reduced, SIN-1 (20 microg) decreased and L-NOARG (150 microg) did not change the EC50 for the DRN or APtN stimulation-induced reduction of incisional pain. We conclude that the antinociceptive effects of L-NOARG or low doses of SIN-1 are independent on the activity of descending pain control pathways travelling via the DLF, but the antinociceptive effect of stimulating electrically the DRN or APtN can be summated to the effect of low dose of SIN-1 or overcome by the high dose of SIN-1.

Topics: Analgesia; Animals; Disease Models, Animal; Dose-Response Relationship, Drug; Efferent Pathways; Electric Stimulation; Enzyme Inhibitors; Hindlimb; Injections, Spinal; Male; Mesencephalon; Molsidomine; Nitric Oxide Donors; Nitroarginine; Pain; Pain Threshold; Raphe Nuclei; Rats; Rats, Wistar; Spinal Cord

To assess the systemic and nociceptive effects of nitric oxide synthase (NOS) inhibitors in the modulation of acute pain in rats subjected to the formalin test.. Formalin 5% was injected in the hind paw in the presence and absence of NOS inhibitors (e.g., 7-nitro indazole, N-nitro-L-arginine and aminoguanidine). Catheters were chronically implanted to continuously record mean arterial blood pressure (MAP) and heart rate (HR). MAP, HR and paw lifting time were recorded at control and every five minutes for 35 min following formalin and NOS inhibitors.. Formalin injected into the rat hind paw induced a biphasic nociceptive behaviour: an initial acute phase (phase 1: during zero to five minutes after the formalin injection) followed by a prolonged tonic response (phase 2: beginning about ten minutes after the formalin injection). Aminoguanidine, an inhibitor of the inducible NOS and 7-nitro indazole, an inhibitor of the neuronal NOS, did not affect phase 1, whereas N-nitro-L-arginine, a non-selective NOS inhibitor decreased it (49%). All three NOS inhibitors diminished nociceptive behaviours during phase 2. L-arginine reversed antinociceptive effects of N-nitro-L-arginine in phase 1 and in phase 2. Pressor effects induced by formalin in phase 1 were abolished following all three NOS inhibitors. During phase 2, formalin-induced pressor effects remained unaffected by N-nitro-L-arginine and aminoguanidine but were inhibited by 7-nitro indazole.. Our data demonstrate that NO is predominantly generated by vascular endothelial NOS in phase 1 and phase 2, whereas the neuronal NOS and the inducible NOS exhibit antinociceptive effects through a non-NO related pathway in phases 1 and 2 in rats subjected to the formalin test.

Topics: Analgesics; Animals; Arginine; Blood Pressure; Enzyme Inhibitors; Formaldehyde; Guanidines; Heart Rate; Indazoles; Nitric Oxide Synthase; Nitroarginine; Pain; Pain Measurement; Rats; Rats, Sprague-Dawley; Sodium Chloride; Time Factors

In view of the scarce information about the analgesic mechanism of kappa-opioid receptor agonists, the objective of the present study was to determine whether nitric oxide (NO) is involved in the peripheral antinociception of bremazocine, a kappa-opioid receptor agonist. Three drugs all interfering with the L-arginine/NO/cyclic GMP pathway were tested using the rat paw model of carrageenan-induced (250 microg) hyperalgesia: (a) N(G)-nitro-L-arginine (a nonselective NO-synthase inhibitor), (b) methylene blue (a guanylate cyclase inhibitor), and (c) zaprinast (a cyclic GMP phosphodiesterase inhibitor). Intraplantar administration of bremazocine (20, 40 and 50 microg) caused a dose-dependent peripheral antihyperalgesia against carrageenan-induced hyperalgesia. The possibility of the higher dose of bremazocine (50 microg) having central or systemic effect was excluded since administration of the drug into the left paw did not elicit antinociception in the contralateral paw. However, when the dose of bremazocine was increased to 100 microg, a significant increase in the nociceptive threshold was observed, as measured in the hyperalgesic contralateral paw. Peripheral antihyperalgesia induced by bremazocine (50 microg) was significantly reduced in a dose-dependent manner when N(G)-nitro-L-arginine (6, 9, 12 and 25 microg) or methylene blue (250, 375 and 500 microg) was injected before. Previous treatment with 50 microg of zaprinast (which had no effect when administered alone) potentiated the antihyperalgesic effect of bremazocine (20 microg). Our data suggest that bremazocine elicits peripheral antinociception by activation of the L-arginine/NO/cyclic GMP pathway and that nitric oxide is an intermediary in this mechanism, forming cyclic GMP.

Topics: Analgesics; Animals; Arginine; Benzomorphans; Cyclic GMP; Guanylate Cyclase; Hyperalgesia; Male; Methylene Blue; Nitric Oxide; Nitric Oxide Synthase; Nitroarginine; Pain; Phosphodiesterase Inhibitors; Purinones; Rats; Rats, Wistar; Receptors, Opioid, kappa

This study evaluates the actions of the new ruthenium complexes trans-[RuCl2(nic)4] (Complex I) and trans-[RuCl2(i-nic)4] (Complex II) as antinociceptives, and their interaction with nitric oxide isoenzymes and with acetylcholine-induced relaxation of rat and rabbit aorta. Complex II inhibited, in a graded manner, neuronal and inducible nitric oxide (NO) synthase, and was about two fold more effective in inhibiting the neuronal NO synthase than the inducible form of the enzyme. Complex I was inactive. Both complexes failed to interfere with constitutive endothelial nitric oxide synthase because they did not change the mean arterial blood pressure of rats. The vasorelaxant effect of acetylcholine was markedly antagonised by the Complexes I and II in rings of both rat and rabbit aorta. Complexes I and II, given intraperitoneally, like N(omega)-nitro-L-arginine methyl ester (L-NAME) and N(G)-nitro-L-arginine (L-NOARG), inhibited, in a graded manner, both phases of the pain response induced by formalin. The actions of L-NAME, L-NOARG and Complex II, but not that of Complex I, were largely reversed by L-arginine. Both complexes failed to affect the motor response of animals in the rota-rod test and had no effect in the hot-plate assay. Together, these findings provide indications that the new ruthenium complexes, especially Complex II and its derivatives, might be of potential therapeutic benefit in the management of pain disorders.

Topics: Analgesics; Animals; Blood Pressure; Drug Interactions; Enzyme Inhibitors; Isoenzymes; Male; Mice; Motor Activity; Muscle, Smooth, Vascular; NG-Nitroarginine Methyl Ester; Nitric Oxide Synthase; Nitroarginine; Pain; Rabbits; Rats; Rats, Wistar; Ruthenium Compounds; Structure-Activity Relationship

Twice daily injections of L-arginine (200 mg/kg, i.p.), a precursor for nitric oxide (NO), for 4 days decreased morphine antinociception in male Swiss-Webster mice. Chronic treatment with L-arginine also produced significant decreases in morphine levels in midbrain, pons and medulla, hippocampus, corpus striatum and spinal cord of mice following an injection of morphine (10 mg/kg, s.c.) in comparison to vehicle-injected mice. N(G)-nitro-L-arginine (L-NNA), an inhibitor of NO synthase (NOS), (5 mg/kg, i.p.) given prior to each injection of L-arginine reversed the effects of the latter on morphine antinociception and decreases in morphine levels in brain regions and spinal cord. Chronic injections of L-NNA alone did not modify either morphine antinociception or morphine distribution in brain regions and spinal cord of mice. These results suggest that decreases in morphine antinociception by chronic treatment with L-arginine is related to the decreases in the entry of morphine in the central sites. The reversal of L-arginine-induced effects by L-NNA suggests that NO-NOS system may be playing a critical role in the regulation of blood-brain barrier to morphine.

Topics: Analgesics, Opioid; Animals; Arginine; Brain; Male; Mice; Morphine; Nitroarginine; Pain; Spinal Cord; Tissue Distribution

Local application of nicotine over the surface of the left ventricle and also occlusion of the left anterior descending coronary artery in the lightly anaesthetised, open-chested, artificially ventilated cat resulted a biphasic rectal movement--initial relaxation followed by sustained contraction. However, distension of the atrial appendage did not evoke any change in rectal motility, indicating the non-involvement of atrial volume receptors in initiating this rectal response of cardiac origin. The relaxation phase of this response was not abolished by pretreatment with atropine or with phentolamine or propranolol but was abolished by the nitric oxide inhibitor, N(G)-nitro-L-arginine (LNNA), and this blockade of the relaxation phase by LNNA was reversed by L-arginine. The contraction phase, however, was abolished by atropine. From these observations it is clear that the relaxation phase of the rectal response to coronary occlusion or epicardial nicotine is mediated through neither cholinergic nor adrenergic pathways but through the release of nitric oxide whereas the contraction phase of such a cardio-rectal response is mediated through the release of the neurotransmitter, acetylcholine.

Topics: Acetylcholine; Afferent Pathways; Animals; Arginine; Atropine; Blood Pressure; Cats; Coronary Vessels; Female; Ganglionic Blockers; Heart; Hexamethonium; Male; Muscle Contraction; Muscle Relaxation; Myocardial Ischemia; Neurotransmitter Agents; Nicotine; Nitric Oxide; Nitroarginine; Pain; Phentolamine; Propranolol; Rectum

The effects of low-dose x-rays on mouse nociceptive behavior were examined using a formalin injected test that rated the amount of time the animals spent licking the injected hind paw. Male ICR White Swiss mice showed a marked suppression of licking behavior after repeated low-dose x-irradiation (5 cGy/day, 6 consecutive days). The most profound effect was observed on the day 30 after irradiation. The decline of licking behavior, however, was not observed at all following olfactory bulbectomy or vomeronasal tract cut. The analgesic effects could be observed in writhing animals administered acetic acid intraperitoneally. Moreover, analgesia was totally blocked by the administration of N-nitro-L-arginine, a nitric oxide synthase inhibitor, to accessory olfactory bulbs prior to the exposure. The present results indicate that the olfactory system plays an important role in modulation of radiation-induced analgesia, and a possible involvement of nitric oxide in the formation of recognition memory subjected to repeated x-rays. Relatively higher doses (5 cGy x 9 days, 5 cGy x 12 days), however, did not induce such effects, namely, the decline of nociceptive response was limited to the animals irradiated with the smaller dose.

Topics: Analgesia; Animals; Behavior, Animal; Dose-Response Relationship, Radiation; Enzyme Inhibitors; Male; Mice; Mice, Inbred ICR; Microinjections; Nitric Oxide Synthase; Nitroarginine; Nociceptors; Olfactory Pathways; Pain

1. We examine some of the mechanisms underlying the analgesic effects of the hydroalcoholic extracts (HE) of Phyllanthus urinaria and P. niruri against formalin-induced nociception in mice. In addition, we also investigate the action of both HEs against capsaicin-mediated pain. 2. Both prazosin and yohimbine (0.15 mg/kg, i.p.) induced a marked inhibition of the analgesic effect caused by phenylephrine (10 mg/kg, i.p.) and clonidine (0.1 mg/kg, i.p.), respectively, but had no effect on the antinociceptive action caused by HE of P. urinaria (10 mg/kg, i.p.) or P. niruri (30 mg/kg, i.p.). 3. NG-nitro-L-arginine (L-NOARG, 75 mg/kg, i.p.) caused marked analgesic effect against the second phase of formalin-induced pain. Treatment of animals with L-arginine (600 mg/kg) completely antagonized the antinociceptive effect of L-NOARG but had no significant effect against the HE of P. urinaria (10 mg/kg, i.p.) or P. niruri (30 mg/kg. i.p.) analgesic properties. 4. The antinociceptive effects caused by the HEs of P. urinaria (10 mg/kg, i.p.) and P. niruri (30 mg/kg, i.p.) were unaffected by methysergide (5 mg/kg, i.p.), p-chloro-phenylalanine-methyl-ester (100 mg/kg, i.p., once a day for 4 consecutive days) or after previous adrenalectomy of animals. 5. The HE of P. urinaria and P. niruri given either intraperitoneally (1-30 mg/kg) or orally (25-200 mg/kg) caused marked and dose-related inhibition of capsaicin-induced pain with ID50 of 2.1 and 6.1 mg/kg given intraperitoneally and 39 and 35 mg/kg given orally, respectively.

Topics: Adrenergic alpha-Agonists; Adrenergic alpha-Antagonists; Analgesics; Animals; Arginine; Capsaicin; Clonidine; Male; Mice; Nitroarginine; Pain; Phenylephrine; Plant Extracts; Prazosin; Yohimbine

Incubation of slices of neonatal rat spinal cord with nitric oxide donor compounds produced marked elevations in cyclic guanosine 3',5' monophosphate (cGMP) levels. The excitatory amino acid receptor agonists N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) produced smaller increases, which were blocked by the nitric oxide synthase (NOS) inhibitor L-NG-nitroarginine (NOArg), indicating that these cGMP responses were mediated by nitric oxide. Immunocytochemistry revealed that, in response to NMDA, cGMP accumulated in a population of small cells and neuropil in laminae II and III of the dorsal horn. This area was also shown, by reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry, to contain NOS. These observations suggest that, in the rat spinal cord, NMDA receptor activation is linked to the formation of NO and, hence, of cGMP. This pathway is located selectively in the superficial dorsal horn, consistent with a role in the processing of nociceptive signals.

Topics: alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; Amino Acid Oxidoreductases; Animals; Animals, Newborn; Arginine; Biomarkers; Cyclic GMP; Female; Glutathione; Male; N-Methylaspartate; NADPH Dehydrogenase; Nerve Tissue Proteins; Nitric Oxide; Nitric Oxide Synthase; Nitroarginine; Nitroprusside; Nitroso Compounds; Pain; Rats; Rats, Wistar; Receptors, N-Methyl-D-Aspartate; S-Nitrosoglutathione; Second Messenger Systems; Signal Transduction; Spinal Cord

This study evaluated the role of nitric oxide (NO) in spinal cord nociceptive transmission during peripheral inflammation evoked by formalin injection into the rat paw, using N omega-nitro-L-arginine (N-Arg), an NO synthase inhibitor. Male rats were prepared with intrathecal (IT) catheters. To quantify the formalin response, the instances of "flinching behavior" were counted at 5-min intervals for 60 min. IT N-Arg depressed the flinching behavior in a dose-dependent manner when N-Arg was administered 10 min before the formalin injection. This N-Arg effect was reversed with L-arginine but not D-arginine. We conclude that NO plays an important role in nociceptive transmission in the spinal cord during the formalin test.

Topics: Amino Acid Oxidoreductases; Animals; Arginine; Dose-Response Relationship, Drug; Formaldehyde; Inflammation; Injections, Spinal; Male; Nitric Oxide Synthase; Nitroarginine; Pain; Rats; Rats, Sprague-Dawley; Spinal Cord

The effects of N-methyl-D-aspartate (100 amol-1 nmol) on the nociceptive tail-flick reflex were studied in awake rats. Lesser doses of N-methyl-D-aspartate (100 amol-10 pmol) administered intrathecally to the lumbar spinal cord produced a dose-dependent facilitation of the tail-flick reflex (maximum at 0.5-1 min). The greatest dose tested (1 nmol) inhibited the tail-flick reflex (maximum at 2-5 min) and produced a caudally directed scratching and biting behavior accompanied by vocalizations. Intrathecal pretreatment with the N-methyl-D-aspartate receptor antagonist, D-2-amino-5-phosphonovaleric acid (1 fmol-1 pmol), which produced no change in baseline tail-flick latency, blocked all N-methyl-D-aspartate produced effects in a dose-dependent manner (100 fmol D-2-amino-5-phosphonovaleric acid produced maximum blockage for about 40 min). The magnitude and duration of N-methyl-D-aspartate-produced biphasic effects on tail-flick latency were similar in awake and lightly pentobarbital-anesthetized rats, but caudally directed biting and scratching behavior was not produced in lightly anesthetized rats. Reversible spinalization at T8-T10 in lightly anesthetized rats (produced by cold-block) completely abolished inhibition of the tail-flick reflex produced by 1 nmol N-methyl-D-aspartate whereas facilitation produced by 10 pmol N-methyl-D-aspartate remained unchanged, indicating that N-methyl-D-aspartate-produced facilitation is a local, segmental effect and that N-methyl-D-aspartate-produced inhibition requires a supraspinal loop. To examine the nature of the supraspinal loop, potential contributions of descending noradrenergic and serotonergic systems were studied. Intrathecal pretreatment with 100 nmol phentolamine completely blocked N-methyl-D-aspartate-produced inhibition of the tail-flick reflex, while N-methyl-D-aspartate-produced facilitation and caudally directed biting and scratching behavior remained unchanged. Intrathecal pretreatment with 50 nmol methysergide reversed the inhibitory effect of 1 nmol N-methyl-D-aspartate, resulting in a potent and prolonged facilitation which could be blocked by D-2-amino-5-phosphonovaleric acid. (1 pmol). Intrathecal pretreatment with an alternate substrate for nitric oxide synthase, NG-nitro-L-arginine methyl ester (100 nmol), completely blocked N-methyl-D-aspartate-produced facilitation of the tail-flick reflex, whereas N-methyl-D-aspartate-produced inhibition and caudally directed biting and scratching behavior were

Topics: 2-Amino-5-phosphonovalerate; Animals; Arginine; Behavior, Animal; Cold Temperature; Hot Temperature; Male; Methysergide; N-Methylaspartate; Nitroarginine; Nociceptors; Pain; Phentolamine; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate; Reflex; Spinal Cord