Increase In Hot Plate Latency Research Articles

Sodium nitrate preconditioning prevents progression of the neuropathic pain in streptozotocin-induced diabetes Wistar rats.

The purpose of the study was to evaluate the possible protective effects of low dose sodium nitrate preconditioning on the peripheral neuropathy in streptozotocin (STZ)-induced diabetic model. Male Wistar rats were randomly divided into five groups: control (no intervention), control treated sodium nitrate (100mg/L in drinking water), diabetic (no intervention), diabetic treated NPH insulin (2-4U), and diabetic treated sodium nitrate (100mg/L in drinking water). Diabetes was induced by intraperitoneal injection of STZ (60mg/kg). All interventions were done for 60days immediately following diabetes confirmation. Thermal and mechanical algesia thresholds were measured by means of hot-plate test, von Frey test, and tail-withdrawal test before the diabetic induction and after diabetes confirmation. At the end of the experiment, serum NOx level and serum insulin level were assessed. Blood glucose concentration and body weight have recorded at the base and duration of the experiment. Both hypoalgesia, hyperalgesia along with allodynia developed in diabetic rats. Significant alterations including, decrease in tail withdrawal latency (30th day), decreased mechanical threshold (60th day), and an increase in hot plate latency (61st day) were displayed in diabetic rats compared to control rats. Nitrate and insulin preconditioning produced protective effects against diabetes-induced peripheral neuropathy. Data analysis also showed a significant increase in glucose level as well as a considerable reduction in serum insulin and body weight of diabetic rats, which restored by both insulin and nitrate preconditioning. Sodium nitrate preconditioning produces a protective effect in diabetic neuropathy, which may be mediated by its antihyperglycemic effects and increased serum insulin level.

Enhanced antinociception with repeated microinjections of apomorphine into the periaqueductal gray of male and female rats.

Dopamine neurons in the ventrolateral periaqueductal gray (PAG) have been reported to contribute to antinociception. The objective of this study was to determine how this dopamine-mediated antinociception differs from what is known about morphine-induced antinociception. Microinjection of the dopamine receptor agonist apomorphine into the PAG produced a dose-dependent increase in hot plate latency and a decrease in open field activity that was greater in male than in female rats. The peak antinociceptive effect occurred 5 min after apomorphine administration. Surprisingly, the antinociceptive potency of apomorphine was enhanced following systemic administration of the opioid receptor antagonist naloxone in male, but not in female rats. The antinociceptive potency of microinjecting apomorphine into the ventrolateral PAG in male and female rats was also enhanced following twice-daily injections for 2 days. The characteristics of apomorphine-induced antinociception differ from previous reports of morphine antinociception following PAG microinjections in that morphine antinociception peaks at 15 min, is blocked by naloxone, and is susceptible to tolerance with repeated administration. These results indicate that apomorphine-induced antinociception is distinct from opioid-induced antinociception, and that dopamine receptor agonists may provide a novel approach to pain modulation.

Evaluation of Anti-Nociceptive and Anti-Inflammatory Effect of Luteolin in Mice

Flavonoids are polyphenolic compounds that not only impart coloration to plants and fruits, but also protect plants from pathogens, radiation, etc. They serve as a nutrient to plants and possess immense anti-oxidant properties. Research has shown that they exhibit anti-inflammatory, anti-cancer, and neuroprotective properties in both in vitro and in vivo analyses. Luteolin is one such flavonoid belongs to the group of flavones present in herbs such as thyme, chamomile, celery, and green pepper. The epidemiological data on luteolin consumption show that luteolin has anti-inflammatory activity and protects from diseases associated with inflammation. The present study assessed the anti-nociceptive properties of luteolin, a potent anti-inflammatory agent, in mice. The results demonstrated that luteolin produces a significant and dose-dependent increase in hot plate latency and tail withdrawal time. It also reduced the number of abdominal constrictions and paw licking induced by acetic acid and glutamate, respectively. Luteolin inhibited the nociceptive responses in both phases of formalin test. The anti-inflammatory property of luteolin was also confirmed with different anti-inflammatory mice models induced by carrageenan and air pouch. Behavioral changes in luteolin-treated mice were assessed with open-field test to confirm the muscle relaxant property. The results of the current study from various pain and inflammatory models confirms that luteolin possess potent anti-nociceptive and anti-inflammatory properties and can thus be used as a drug in pain management.

Functionally selective signaling for morphine and fentanyl antinociception and tolerance mediated by the rat periaqueductal gray.

Functionally selective signaling appears to contribute to the variability in mechanisms that underlie tolerance to the antinociceptive effects of opioids. The present study tested this hypothesis by examining the contribution of G protein-coupled receptor kinase (GRK)/Protein kinase C (PKC) and C-Jun N-terminal kinase (JNK) activation on both the expression and development of tolerance to morphine and fentanyl microinjected into the ventrolateral periaqueductal gray of the rat. Microinjection of morphine or fentanyl into the periaqueductal gray produced a dose-dependent increase in hot plate latency. Microinjection of the non-specific GRK/PKC inhibitor Ro 32-0432 into the periaqueductal gray to block mu-opioid receptor phosphorylation enhanced the antinociceptive effect of morphine but had no effect on fentanyl antinociception. Microinjection of the JNK inhibitor SP600125 had no effect on morphine or fentanyl antinociception, but blocked the expression of tolerance to repeated morphine microinjections. In contrast, a microinjection of Ro 32-0432 blocked the expression of fentanyl, but not morphine tolerance. Repeated microinjections of Ro 32-0432 blocked the development of morphine tolerance and inhibited fentanyl antinociception whether rats were tolerant or not. Repeated microinjections of SP600125 into the periaqueductal gray blocked the development of tolerance to both morphine and fentanyl microinjections. These data demonstrate that the signaling molecules that contribute to tolerance vary depending on the opioid and methodology used to assess tolerance (expression vs. development of tolerance). This signaling difference is especially clear for the expression of tolerance in which JNK contributes to morphine tolerance and GRK/PKC contributes to fentanyl tolerance.

Opioid Selective Antinociception Following Microinjection Into the Periaqueductal Gray of the Rat

Morphine and fentanyl produce antinociception in part by binding to mu-opioid receptors in the periaqueductal gray (PAG). The present study tested the hypothesis that the PAG also contributes to the antinociceptive effects of other commonly used opioids (oxycodone, methadone, and buprenorphine). Microinjection of high doses of oxycodone (32–188 μg/.4 μL) into the ventrolateral PAG of the rat produced a dose-dependent increase in hot plate latency. This antinociception was evident within 5 minutes and nearly gone by 30 minutes. In contrast, no antinociception was evident following microinjection of methadone or buprenorphine into the ventrolateral PAG despite use of a wide range of doses and test times. Antinociception was evident following subsequent microinjection of morphine into the same injection sites or following systemic administration of buprenorphine, demonstrating that the injections sites and drugs could support antinociception. Antinociception to systemic, but not PAG, administration of buprenorphine occurred in both male and female rats. These and previous data demonstrate that the mu-opioid receptor signaling pathway for antinociception in the PAG is selectively activated by some commonly used opioids (eg, morphine, fentanyl, and oxycodone) but not others (eg, methadone or buprenorphine). The fact that methadone and buprenorphine produce antinociception following systemic administration demonstrates that mu-opioid receptor signaling varies depending on location in the nervous system. PerspectiveThis study demonstrates that the PAG contributes to the antinociceptive effects of some commonly used opioids (morphine, fentanyl, and oxycodone) but not others (methadone or buprenorphine). Such functional selectivity in PAG-mediated opioid antinociception helps explain why the analgesic profile of opioids is so variable.

Effect Of Buspirone On Inflammation, Pain And Gastric Injury In Mice

Buspirone, a 5HT1A receptor partial agonist, was evaluated in various acute nociceptive pain models, on the carrgaeenaninduced paw oedema, in Porsolt's forced-swimming test, on haloperidol-induced catalepsy, on the indomethacin-induced gastric mucosal damage and on gastric acid secretion in mice. Buspirone (2, 4 or 8 mg/kg, i.p.) significantly increased the response latency in the mouse hot plate test. The anti-nociceptive activity was observed with 2 mg/kg and a maximal increase in hot-plate latency by 41.3%. Buspirone in addition markedly increased the nociceptive thresholds in the acetic-acid-induced writhing assay, in the capsaicin-induced chemogenic pain and in electrically-induced pain in mice. It displayed no antidepressant activity in the Porsolt's forced-swimming test, but at doses of 8 or 16 mg/kg, buspirone decreased the duration of catalepsy induced by haloperidol by 28.1 and 58.3%, respectively. When given 30 min prior to subplantar carrageenan injection, the drug at 8, 16 mg/kg, inhibited paw oedema response 4 h post-carrageenan by 26.3, 41.6%, respectively. Buspirone (2, 4 or 8 mg/kg, i.p.) significantly inhibited the development of indomethacin gastric mucosal lesions and gastric acid secretion in a dose-dependent manner. In conclusion, Buspirone increased pain threshold in models of thermal, chemogenic, electrical and visceral pain and displayed anti-inflammatory and gastric protective properties. The drug might prove of value for the management of inflammatory painful conditions.

Antinociceptive Effects of the Aqueous Extract of Brugmansia suaveolens Flowers in Mice

The infusion of Brugmansia suaveolens, popularly known as trombeteira or cartucheira, has been used to treat pain in Brazil. The present study was conducted to test for its antinociceptive effects using the abdominal-writhing, formalin, tail-flick, and hot-plate tests in mice. The aqueous extract from B. suaveolens flowers administered intraperitoneally at doses of 100 and 300 mg/kg body weight significantly inhibited acetic acid-induced abdominal constrictions. An increase in hot-plate latency was also observed in animals receiving both doses (100 and 300 mg/kg). In the formalin test, both doses from the aqueous extract inhibited the first (0-5 min) and second phase (20-25 min). Tail-flick assays demonstrated that treatment of animals with plant extract induced attenuation of the response. These results suggest that the aqueous extract from B. suaveolens flowers produced antinociceptive effects, as demonstrated in the experimental models of nociception in mice. This supports popular medicinal uses of this plant as an analgesic.

The involvement of nitric oxide on the analgesic effect of tramadol

The involvement of nitric oxide on the analgesic effect of tramadol

Intermittent dosing prolongs tolerance to the antinociceptive effect of morphine microinjection into the periaqueductal gray

Tolerance to the antinociceptive effect of microinjecting morphine into the ventrolateral periaqueductal gray (vPAG) develops with repeated administration. The objective of the present experiment was to determine whether the magnitude and duration of tolerance differ depending on the interval between injections. Rats were injected with morphine or saline into the vPAG either once daily for 4 days or twice daily for 2 days. All rats were injected with morphine into the vPAG for the fifth injection to determine whether tolerance had developed. Morphine microinjection produced tolerance in both morphine-pretreated groups regardless of inter-dose interval. One and two weeks later, microinjection of morphine produced an increase in hot plate latency in all groups except rats pretreated with daily morphine microinjections. That is, tolerance was evident 2 weeks following the induction of tolerance produced by daily, but not twice daily injections of morphine. Although a long inter-dose interval has been shown to prolong the duration of tolerance after systemic morphine administration, this is the first report showing a similar effect with direct administration of morphine into the brain. Given that associative learning underlies prolonged tolerance with systemic morphine administration, the present data suggest that associative mechanisms of tolerance are also engaged when morphine administration is restricted to the PAG.

Defensive behaviors evoked from the ventrolateral periaqueductal gray of the rat: Comparison of opioid and GABA disinhibition

Microinjection of morphine into the ventrolateral periaqueductal gray (PAG) disinhibits output neurons resulting in immobility and antinociception. Disinhibition also can be produced by microinjection of the GABA antagonist bicuculline. If morphine and bicuculline disinhibit the same class of neurons, then the behavioral effects evoked should be the same. Microinjection of morphine (5 μg/0.4 μl) into the ventrolateral PAG produced antinociception in 46 of 85 rats (54%). Subsets of rats with and without morphine antinociception were subsequently injected with bicuculline (2.5, 5, 10, and 25 ng/0.4 μl) into the same PAG site. Microinjection of bicuculline produced an increase in hot plate latency that was independent of the effect of the prior morphine microinjection. Bicuculline administration also produced an increase in locomotor activity in most rats, not immobility as with morphine microinjections. These differences between morphine and bicuculline microinjections indicate three things: (a) disinhibition of PAG neurons whether by morphine or bicuculline is an effective means of producing antinociception; (b) the circuitry underlying the behavioral effects of morphine and bicuculline differ; (c) the ventrolateral PAG appears capable of supporting a range of defensive behaviors from immobility to flight.

Effects of N-Methyl-D-Aspartate Receptor Antagonists on Acute Morphine-Induced and l-Methadone-Induced Antinociception in Mice

Although N-methyl-D-aspartate (NMDA) receptor antagonists clearly attenuate the development of tolerance to the antinociceptive effects of opioids, it is not clear whether they also alter acute opioid-induced antinociception. The present study was designed to assess NMDA/opioid interactions in C57BL/6 mice by examining various NMDA receptor antagonists of different selectivity in combination with the μ opioid receptor agonists morphine and l-methadone. A mouse hot plate procedure was used to assess the effects of morphine (0.1 to 10.0 mg/kg) and l-methadone (0.1 to 5.6 mg/kg) alone and after pretreatment with the competitive NMDA receptor antagonist LY235959 (0.1 to 1.0 mg/kg), the glycine site NMDA receptor antagonist R(+)-HA-966 (10.0 to 56.0 mg/kg), or the polyamine site and NR2B selective NMDA receptor antagonist ifenprodil (3.2 to 10.0 mg/kg). Morphine and l-methadone produced dose- and time-dependent increases in 56°C hot plate latencies. At the doses tested, the NMDA receptor antagonists produced no effect on hot plate latencies. However, when these drugs were combined with morphine, latency to respond to the hot plate was significantly increased from morphine alone. Combinations of the NMDA receptor antagonist LY235959 and l-methadone produced similar increases in hot plate latencies; however, combinations of l-methadone with R(+)-HA-966 or ifenprodil did not increase hot plate latencies compared with l-methadone alone. These results suggest that a range of NMDA receptor antagonists potentiate morphine-induced antinociception, although the potentiation of l-methadone might be specific to the antagonist examined. Perspective The inclusion of low-dose NMDA receptor antagonists to opioids might be beneficial for the treatment of acute pain by enhancing the antinociceptive effects of the opioid.

Anti-Inflammatory, Antinociceptive, and Gastric Effects ofHypericum perforatumin Rats

The pharmacological activity of Hypericum perforatum was assessed using models of inflammation, nociception, and gastric mucosal injury in rats. H. perforatum was given systemically as well as orally. When administered systemically, H. perforatum (50–300 mg/kg, s.c.) produced a dose-related and significant inhibition of the edematogenic response to s.p. injection of carrageenan. The percentages of maximal inhibition by the above doses were 53.7, 61.3, and 75.3%, respectively (compared to 90% after 50 mg/kg fluoxetine and 60.7% after 72 mg/kg etodolac). In tests of nociception, H. perforatum, administered orally, displayed antinociceptive activity in the tail electric stimulation and hot plate tests. The antinociceptive activity was observed with 25 mg/kg and a maximal increase in hot plate latency by 50% (compared to 73.2 and 77.8% increases by 5 or 10 mg/kg fluoxetine, respectively). In contrast, the acetic acid–induced (0.6%, i.p.) writhing was significantly reduced by fluoxetine or etodolac, but not H. perforatum. Also, the nociceptive response caused by i.p. injection of capsaicin (1.6 μg/paw) was unaffected by H. perforatum, but reduced by fluoxetine. Injection of H. perforatum (50, 125, or 250 mg/kg, s.c.) to pylorus-ligated rats, decreased gastric acid secretion, but increased indomethacin-induced gastric mucosal lesions dose dependently. These results demonstrate that H. perforatum exhibits antiedematogenic and antinociceptive properties, which may be of value for the management of inflammatory painful conditions. The agent, however, causes gastric irritation and may aggravate that of NSAIDs.

Behavioral and electrophysiological evidence for tolerance to continuous morphine administration into the ventrolateral periaqueductal gray

Repeated microinjections of morphine into the ventrolateral periaqueductal gray (vPAG) produce tolerance to the antinociceptive effect of morphine [Behav Neurosci 113 (1999) 833]. These results may be a direct effect of morphine on cells within the vPAG or be caused by cues linked to the microinjection procedure (i.e. associative tolerance). The objective of this paper was to determine whether continuous administration of morphine into the vPAG (i.e. no cues) would produce tolerance. Tolerance was assessed by measuring changes in behavior and changes in the activity of neurons in the rostral ventromedial medulla (RVM), the primary output target of the PAG. Rats were implanted with an osmotic minipump that released morphine (2.5 or 5 μg/h) or saline into the vPAG continuously. Continuous administration of morphine produced an increase in hotplate latency when measured 6 h after initiation of treatment. Tolerance to this antinociception was evident within 24 h. After 3 days, rats were anesthetized and the activity of RVM neurons was assessed. Although acute morphine administration into the RVM inhibits the activity of RVM on-cells and enhances the activity of off-cells, these neurons appeared normal following 3 days of continuous morphine administration. Systemic naloxone administration produced hyperalgesia that was associated with a marked increase in on-cell activity and a complete cessation of off-cell activity. The loss of morphine inhibition of nociception, measured behaviorally and electrophysiologically, demonstrates that tolerance is caused by a direct action of morphine on vPAG neurons.

The effects of deleting the mouse neurotensin receptor NTR1 on central and peripheral responses to neurotensin.

Mice deficient in the neurotensin (NT)-1 receptor (NTR1) were developed to characterize the NT receptor subtypes that mediate various in vivo responses to NT. F2 generation (C57BL6/Sv129J) NTR1 knockout (-/-) mice were viable, and showed normal growth and overt behavior. The -/- mice lacked detectable NTR1 radioligand binding in brain, whereas NTR2 receptor binding density appeared normal compared with wild-type (+/+) mice. The gene deletion also resulted in the loss of NTR1 expression as determined by reverse transcription-polymerase chain reaction and in situ hybridization. Intracerebroventricular injection of NT (1 microg) to +/+ mice caused a robust hypothermic response (5-6 degrees C) and a significant increase in hot-plate latency. These effects were absent in the -/- mice. Similar results were obtained with i.p. injections of the brain-penetrant NT analog NMe-Arg-Lys-Pro-Trp-Tle-Leu (NT-2, 1 mg/kg i.p.). NT-2 administration also impaired rotarod performance in wild-type mice, but had no effect on motor coordination in knockout mice. In vitro, NT and NT-2 at 30 nM caused predominantly contraction and relaxation in isolated distal colon and proximal ileum, respectively, from +/+ mice, but no responses were observed with tissues from -/- mice. A similar loss of the contractile effects of NT was observed in the isolated stomach fundus from the knockout mice. In vivo, NT-2 administration reduced colonic propulsion substantially in wild-type mice. In contrast, NT-2 had no effect in NTR1 null mice, whereas the hypomotility effect of clonidine was intact. These data indicate that NTR1 mediates several of the central and peripheral effects of NT.

Antinociceptive and antiedematogenic effects of the aqueous extract of Hyptis pectinata leaves in experimental animals

The aqueous leaf extract of Hyptis pectinata (L.) Poit (Lamiaceae), popularly known in Brazil as “sambaicatá” or “canudinho”, was tested for its antinociceptive effects using the abdominal writhing, hot plate and formalin test models, and for its aniedematogenic effects using the carrageenin and arachidonic acid-induced rat paw edema. The aqueous extract of Hyptis pectinata administered orally at doses of 100, 200 and 400 mg/kg had a significant antinociceptive effect in the test of acetic acid-induced abdominal writhing, with 43, 51 and 54% reduction of writhes, respectively, compared to the control. An increase in hot-plate latency of 47 and 37.5% was also observed in animals receiving doses of 200 and 400 mg/kg, p.o. when placed on a hot plate. In the formalin test, doses of 200 and 400 mg/kg, p.o. had no significant effect during the first phase of the test (0–5 min), while the dose of 200 mg/kg, p.o. reduced the nociceptive effect by 70% during the second phase (20–25 min). At the dose of 600 mg/kg, p.o., the aqueous extract inhibited carrageenin-induced rat paw edema by 34.1%, and the dose of 300 mg/kg administered intraperitoneally inhibited the rat paw edema induced by subplantar injection of arachidonic acid by 32.8%. These results suggest that the aqueous extract from the Hyptis pectinata leaves produces antiedematogenic and antinociceptive effects. The antinocipetion observed with the hot-plate test probably involves the participation of the opioid system.

Sensory thresholds and the antinociceptive effects of GABA receptor agonists in mice lacking the β 3 subunit of the GABA A receptor

A line of mice was recently created in which the gabrb3 gene, which encodes the β 3 subunit of the GABA A receptor, was inactivated by gene-targeting. The existence of mice with a significantly reduced population of GABA A receptors in the CNS enabled an investigation of the role of GABA and GABA A receptors in nociception. The present study examined the sensory thresholds of these mice, as well as the antinociceptive effects of subcutaneously or intrathecally administered GABA A and GABA B receptor agonists. Homozygous null (β 3 −/−) mice displayed enhanced responsiveness to low-intensity thermal stimuli in the tail-flick and hot-plate test compared to C57BL/6J and 129/SvJ progenitor strain mice, and their wild-type (β 3 +/+) and heterozygous (β 3 +/−) littermates. The β 3 −/− mice also exhibited enhanced responsiveness to innocuous tactile stimuli compared to C57BL/6J, 129/SvJ and to their β 3 +/+ littermates as assessed by von Frey filaments. The presence of thermal hyperalgesia and tactile allodynia in β 3 −/− mice is consistent with a loss of inhibition mediated by presynaptic and postsynaptic GABA A receptors in the spinal cord. As expected, subcutaneous administration of the GABA A receptor agonist 4,5,6,7-tetrahydroisoxazolo-(5,4-c)pyridin-3-ol did not produce antinociception in β 3 −/− mice, whereas it produced a dose-dependent increase in hot-plate latency in C57BL/6J, 129/SvJ, β 3 +/+ and β 3 +/− mice. However, the antinociceptive effect of the GABA B receptor agonist baclofen in the tail-flick and hot-plate tests was also reduced in β 3 −/− mice compared to the progenitor strains, β 3 +/+ or β 3 +/− mice after either subcutaneous or intrathecal administration. This finding was unexpected and suggests that a reduction in GABA A receptors can affect the production of antinociception by other analgesic drugs as well.

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 microg) 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 microg). In the hot plate test, only the largest dose of ketamine (500 microg) caused a nonstereospecific, 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. 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.

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)

Studies on the safety of chronically administered intrathecal neostigmine methylsulfate in rats and dogs.

The spinal delivery of the cholinesterase inhibitor neostigmine yields analgesia in rats and augments the analgesic effects of alpha 2 agonists in sheep. To assess its activity in humans, preclinical toxicology studies to define its safety were required in two species. Rats with chronic intrathecal catheters received daily injections of saline (vehicle) or 5 micrograms/10 microliters or 10 micrograms/10 microliters neostigmine HCl (n = 6/group) for 4 days and were observed for general behavior and nociception (52.5 degrees C hot plate). On day 6, rats were anesthetized and submitted to whole body perfusion/fixation. For dog studies, male beagles were prepared following rigid aseptic precautions with catheters passed from the cisterna magna to the lumbar intrathecal space. Catheters were connected to an external vest-mounted pump. Based on preliminary studies, ten implanted dogs were randomly assigned to receive infusions of neostigmine for 28 days (4 mg/4 ml/day; n = 6) or saline (4 ml/day; n = 4). At 28 days, dogs were anesthetized, cisternal cerebrospinal fluid was obtained, and dogs were submitted to perfusion-fixation. Rat and dog spinal cords were embedded, sectioned, stained, and assessed by the pathologist without knowledge of treatment. In rats, neostigmine produced a dose-dependent increase in hot plate latency, and no tolerance was observed. Mild tremor was observed but was not debilitating. Histopathology revealed a mild fibrotic reaction to the catheter with mixed signs of moderate, acute, and chronic inflammation with no differences between saline or drug groups. In dogs, neostigmine had no effect on blood pressure or on the skin twitch response but produced bradycardia and an increase in muscle tone. At sacrifice, cerebrospinal fluid protein, specific gravity, and glucose were elevated in both saline and neostigmine groups. Histopathology displayed a local reaction to the spinal catheter and a mixed acute and chronic inflammatory reaction. No group differences were observed. These results suggest that, at the neostigmine concentration of 1 mg/ml in the rat and dog and in doses up to 4 mg/day in the dog, there is no evidence of spinal tissue toxicity that can be attributed to the drug. This result, observed in two species, suggests that intrathecal neostigmine given in this manner is without distinguishable toxicity in these two models.

Age-dependent attenuation of the decrease of C fibers by capsaicin and its effects on responses to nociceptive stimuli.

The effects of subcutaneous treatment of mice 10, 15, or 20 days (young) or 30 or 60 days (adult) of age with capsaicin on development of unmyelinated (C) fibers in the L4 dorsal roots were examined. The responses of the mice 2-4 months later to thermal (hot plate; 55 degrees C) and neurogenic plasma extravasative (chemogenic nociception) stimuli were evaluated. Capsaicin treatment 10 days after birth affected development of myelinated fibers significantly (7.9% reduction). Capsaicin treatment 10, 15, 20, and 30 days after birth reduced the number of C fibers 11.0-51.7% and even treatment 60 days after birth caused a significant decrease (10.0%) in the mean number of C fibers. The destruction of C fibers by capsaicin was attenuated during development, but individual differences in the reduction of C fibers were observed on and after 15 days of life and seemed to become more marked over time. Neurogenic plasma extravasation related to primary sensory neurons was decreased by capsaicin, irrespective of the time of treatment. In parallel with reduction of C fibers, hot-plate latency was increased significantly by treatment of young animals with capsaicin. These results suggest that the effect of capsaicin on thermal nociception is age-dependent and is correlated with decrease of C fibers. However, a marked increase in hot-plate latency did not always correspond to a marked decrease of C fibers. In contrast, the reduction of plasma extravasation was not age-dependent: Reduced extravasation of dye persisted during development.(ABSTRACT TRUNCATED AT 250 WORDS)