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Objective To test the anesthetic effect of a bupivacaine liposome injectable suspension (BLIS), used in a multiple-dose manner for up to 5 consecutive days. Study design Prospective, randomized, experimental study. Animals A total of 30 male and female Sprague–Dawley rats (Rattus norvegicus), aged 97 (75–130) days and weighing 337.2 (219.6–465.9) g, mean (range). Methods Rats were assigned to one of five BLIS vial groups, in which drug was administered from a newly opened vial or 1, 2, 3 and 4 days after the vial was opened. The vials were refrigerated between uses. A 14 gauge needle attached to an injection plug was used to puncture each vial once and was not removed; BLIS was withdrawn from the injection plug in a multiple-dose fashion. A dose rate of 0.4 mL kg−1 was administered subcutaneously into the left pelvic limb paw. Antinociception was evaluated using a paw pressure test on both injected and uninjected paws before (time 0, baseline) and 1, 24, 48 and 72 hours after injection. Results Age of BLIS vial had no significant effect on anesthetic efficacy (p = 0.97). Across all groups, paw withdrawal latency averaged 5.23 ± 0.24 seconds at baseline (before BLIS injection), increased to 16.45 ± 0.65 seconds at 1 hour after BLIS injection, declined to 7.50 ± 0.76 seconds at 24 hours after BLIS injection, and further declined thereafter (p Conclusions and clinical relevance BLIS single-use vials retained efficacy when used up to 5 days in a multiple-dose fashion. Because anesthetic effects declined substantially after 24 hours, multimodal pain management remains important for providing analgesia care.

OBJECTIVE To compare the analgesic effects of two types of spinal manipulation (SM) in acute lumbar radiculopathy (ALR) model rats induced by self-transplantation of autologous nucleus pulposus (ANP), and clarify the therapeutic mechanism. METHODS Totally 108 male Sprague-Dawley rats were randomly divided into 6 groups by a random number table (18 rats in each group), including a blank group with no interference, a sham operation group with a surgery by making a local soft tissue incision on the left side of L5-6 vertebral segment, a model group with ALR of L5 extraforaminal nerve by ANP self-transplantation without other interference, a sham manipulation (SMA) group with simulating physical rotation, as well as a mobilization (MOB) group with simulating low-velocity and variable-amplitude rotation and a manipulation (MAN) group with simulating high-velocity and low-amplitude rotation. The interventions in SMA, MOB, and MAN groups started 1 day after modeling followed by another 5 treatments at days 3, 5, 8, 10 and 12. Rats in the other 3 groups did not receive any special intervention. Behavioral pain tests of 50% mechanical pain withdrawal threshold (50% PWT) and paw withdrawal latency (PWL) were conducted 1 day before operation followed by another 10 tests on days 1-7, 10, 12 and 14. Immunohistochemical expression of nitric oxide synthase (NOS) was investigated on days 5 and 12 after operation. RESULTS After 3 experimental SM interventions, 50% PWT and PWL were higher in the MAN group than the SMA group on days 6 and 7, and higher on days 10, 12 and 14 postoperatively (P

OBJECTIVE To evaluate antinociceptive efficacy and safety of SC buprenorphine hydrochloride administration in African pygmy hedgehogs (Atelerix albiventris). ANIMALS 12 healthy adult hedgehogs (7 males and 5 females). PROCEDURES 3 crossover experimental trials were performed. In the first trial, all 12 hedgehogs were given single SC injections of buprenorphine (0.01 mg/kg [0.0045 mg/lb]), buprenorphine (0.03 mg/kg [0.014 mg/lb]), or saline (0.9% NaCl) solution (0.16 mL/kg [0.073 mL/lb]), and sedation and hind limb thermal withdrawal latency were measured. In the second trial, 6 hedgehogs were given single SC injections of buprenorphine (0.05 mg/kg [0.023 mg/lb]) or saline solution (0.16 mL/kg), and sedation and withdrawal latency were evaluated. In the third trial, 10 hedgehogs were given 3 doses of buprenorphine (0.05 mg/kg, SC, q 24 h) or saline solution (0.16 mL/kg, SC, q 24 h), and food intake and body weight were measured for 6 days. RESULTS For all 3 experimental trials, the sedation score was 0 for all hedgehogs at all assessment times. A single 0.01-mg/kg dose of buprenorphine significantly increased thermal withdrawal latency for 36 hours, and single 0.03- and 0.05-mg/kg doses significantly increased latencies for 48 hours. Increased locomotor activity was noted in a few hedgehogs after administration of the 0.03- and 0.05-mg/kg doses. Daily administration of buprenorphine did not have significant effects on food intake or body weight. CONCLUSIONS AND CLINICAL RELEVANCE SC administration of buprenorphine at single doses of 0.01, 0.03, and 0.05 mg/kg provided safe, long-lasting antinociception in African pygmy hedgehogs without apparent sedative effects.

OBJECTIVERefractory deafferentation pain has been evidenced to be related to central nervous system neuroplasticity. In this study, the authors sought to explore the underlying glucose metabolic changes in the brain after brachial plexus avulsion, particularly metabolic connectivity.METHODSRats with unilateral deafferentation following brachial plexus avulsion, a pain model of deafferentation pain, were scanned by small-animal 2-deoxy-[18F]fluoro-d-glucose (18F-FDG) PET/CT to explore the changes of metabolic connectivity among different brain regions. Thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) of the intact forepaw were also measured for evaluating pain sensitization. Brain metabolic connectivity and TWL were compared from baseline to 1 week after brachial plexus avulsion.RESULTSAlterations of metabolic connectivity occurred not only within the unilateral hemisphere contralateral to the injured forelimb, but also in the other hemisphere and even in the connections between bilateral hemispheres. Metabolic connectivity significantly decreased between sensorimotor-related areas within the left hemisphere (contralateral to the injured forelimb) (p < 0.05), as well as between areas across bilateral hemispheres (p < 0.05). Connectivity between areas within the right hemisphere (ipsilateral to the injured forelimb) significantly increased (p = 0.034). TWL and MWT of the left (intact) forepaw after surgery were significantly lower than those at baseline (p < 0.001).CONCLUSIONSThis study revealed that unilateral brachial plexus avulsion facilitates pain sensitization in the opposite limb. A specific pattern of brain metabolic changes occurred in this procedure. Metabolic connectivity reorganized not only in the sensorimotor area corresponding to the affected forelimb, but also in extensive areas involving the bilateral hemispheres. These findings may broaden our understanding of central nervous system changes, as well as provide new information and a potential intervention target for nosogenesis of deafferentation pain.

Neuropathic pain is clinically unsatisfactorily treated because of unclear mechanisms. The present study aims to explore the concrete mechanisms underlying the alleviation of resveratrol-activated silent information regulator 1 (Sirt1) to chronic constriction injury (CCI)-induced neuropathic pain. CCI surgery was conducted to the unilateral sciatic nerve of male Sprague-Dawley rats to induce neuropathic pain experimentally. Resveratrol with or without miR-182 antagomir were administered to CCI rats via intrathecal catheter. Behavioral tests including paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were conducted to explore mechanical allodynia and thermal hyperalgesia. Western blot, qRT-PCR were used to detect the expression levels of Sirt1, miR-182, and Nav1.7 in CCI dorsal root ganglions (DRGs). CCI rats displayed lower PWT and PWL compared with the sham control. Also, the CCI DRGs displayed lower Sirt1 and miR-182 expression as well as higher Nav1.7 expression, which would be almost reversed by resveratrol treatment for 4 successive days. We also found that miR-182 expression inhibition erased the analgesia effect of resveratrol to CCI–induced neuropathic pain possibly through upregulating Nav1.7 expression. In summary, resveratrol alleviated CCI–induced neuropathic pain, possibly through activating Sirt1 to suppress Nav1.7 expression via upregulating miR-182 expression in CCI DRGs.

Objectives The aim of this study, conducted at the Military Institute of Hygiene and Epidemiology in Warsaw in 2017, was to evaluate the effects of a single (15 min) and repeated (5 times for 15 min) radio-frequency radiation (RFR) exposure of 1800 MHz frequency on the analgesic efficacy of morphine in healthy rats and rats with complete Freund's adjuvant (CFA) induced inflammation. Material and methods Rats were injected intraperitoneally with morphine (MF) in the dose of 8 mg/kg or drug vehicle 15 min before RFR exposure. The authors used the plantar analgesia meter and the radiant heat paw-withdrawal test to assess the pain threshold. Results A single RFR exposure slightly influenced paw withdrawal latency (PWL) in healthy rats in the single exposure baseline group, and influenced PWL, 30 and 60 min after morphine or vehicle injection, in the repeated exposure group. There were differences between the sham-exposed groups (vehicle), 30, 60 and 90 min after injection, both in the single and repeated RFR-exposure groups. The antinociceptive effect of morphine in healthy rats was slightly decreased by RFR exposure at 60 and 90 min, both in the single and repeated exposure groups. The PWL was slightly decreased, both in the single and repeated exposure groups with inflammation (CFA and CFA/MF), at 30, 60 and 90 min, and PWL was increased in the sham-exposed groups (CFA and CFA/MF), both in the single and repeated exposure groups, at 30, 60 and 90 min. The antinociceptive effect of morphine in healthy rats was significantly increased by RFR exposure at 30 min after drug injection in the single exposure group, and increased at 30 and 60 min in the repeated exposure group. Conclusions The authors observed a minor influence of RFR exposure on the antinociceptive effects of morphine in healthy rats after repeated exposures and a statistically significant influence of repeated exposure on morphine mediated antinociceptive effects in the inflammation group. Int J Occup Med Environ Health. 2019;32(4):465-74.

The present study aimed to investigate cerebral metabolic changes in a neuropathic pain model following deafferentation. A total of 24 Sprague-Dawley rats were included for modeling of right brachial plexus avulsion (BPA) through the posterior approach. As nerve injury would cause central sensitization and facilitate pain sensitivity in other parts of the body, thermal withdrawal latency (TWL) of the intact forepaw was assessed to investigate the level of pain perception following BPA-induced neuropathic pain. [Fluorine-18]-fluoro-2-deoxy- d -glucose (18F-FDG) positron emission tomography (PET) was applied to the brain before and after brachial plexus avulsion to explore metabolic changes in neuropathic pain following deafferentation. The TWL of the left (intact) forepaw was significantly lower after BPA than that of baseline (p

The aim of this study was to investigate whether contralateral moxibustion would generate a similar analgesic effect with ipsilateral moxibustion. Contra- and ipsilateral moxibustion were separately applied to Zusanli (ST36) acupoints of inflammatory pain mice. The analgesic effect was evaluated, respectively, by licking/biting time (LBT) of formalin-induced inflammatory pain and thermal withdrawal latency (TWL) of complete Freund’s adjuvant- (CFA-) induced inflammatory pain. For formalin-induced pain, compared with formalin group, the total LBT of ipsi- and contralateral moxibustion reduced in both phase I and phase II, but there was no significant difference between ipsi- and contralateral moxibustion. For CFA-induced inflammatory pain, compared with CFA group, TWL of ipsi- and contra-Moxi groups increased immediately after moxibustion intervention; however there was no obvious difference between ipsi- and contralateral moxibustion at any timepoint. It indicated that contralateral moxibustion had a similar analgesic effect with ipsilateral moxibustion in both formalin- and CFA-induced pain. These results suggest that both ipsi- and contralateral moxibustion could be applied for pain relief.

The aim of the study was to evaluate the analgesic efficacy of tramadol compared to butorphanol administered locally in ventral surface of the hind paw of rats. Prospective, randomized experimental study; twenty-one adult male Wistar rats were selected. Heart rate (beats minute-1), respiratory rate (breaths minute-1), and paw withdrawal latency (onset of radiant heat until paw withdrawal/seconds) were measured prior (T0) and after (T5, T10, T15, T20) intraplantar injection with saline solution 0,9% (group S), butorphanol 1 mg kg-1 (group B), and tramadol 1 mg kg-1 (group T). Shapiro-Wilk normality test and Friedman test were used to analyze the data expressed by median and range. Statistical significance was set at p < 0.05. Statistical analysis of heart rate showed that there were significant differences between groups at different monitoring times. There were no significant differences in respiratory rate after intraplantar injection in any of the treatment groups. The paw withdrawal latency values at T5, T10, and T15 minutes after intraplantar injection in the group B were significantly higher compared to baseline value and to the values of the other groups. The paw withdrawal latency were no significant changes in the measurements of intragroup in S and T. Intraplantar administration of butorphanol provides a good analgesia and significantly increases paw withdrawal latency compared to tramadol. Intraplantar injection of butorphanol could be useful and safe and safe technique to achieve local analgesia for minor surgical procedures in rats.

Osteoarthritic pain has a strong impact on patients’ quality of life. Understanding the pathogenic mechanisms underlying osteoarthritic pain will likely lead to the development of more effective treatments. In the present study of osteoarthritic model rats, we observed a reduction of M-current density and a remarkable decrease in the levels of KCNQ2 and KCNQ3 proteins and mRNAs in dorsal root ganglia (DRG) neurons, which were associated with hyperalgesic behaviors. The activation of KCNQ/M channels with flupirtine significantly increased the mechanical threshold and prolonged the withdrawal latency of osteoarthritic model rats at 3–14 days after model induction, and all effects of flupirtine were blocked by KCNQ/M-channel antagonist, XE-991. Together, these results indicate that suppression of KCNQ/M channels in primary DRG neurons plays a crucial role in the development of osteoarthritic pain.

Objective To investigate the local initiation role of local exosomes of acupoints in acupuncture analgesic effect. Methods Thirty-two rats with adjuvant arthritis were randomly divided into model group (Group CFA), model + electroacupuncture group (Group EA), model + antagonist + electroacupuncture group (Group GW4869 + EA), and model + dimethyl sulfoxide + electroacupuncture group (Group DMSO + EA), with 8 rats in each group. The model rat s of adjuvant arthritis were prepared by intradermal injection of 0.1 mL of Freund's adjuvant complete into the metatarsal of the right posterior foot to induce inflammation. No intervention was given in Group CFA, while electroacupuncture was performed in the other three groups at “Zusānlĭ (足三里 ST 36, bilaterally)” and “Huantiao (环跳 GB 30, bilaterally)” of the rats with the following electroacupuncture parameters: dilatational wave with a frequency of 2/10 Hz, an intensity of 5/10/15 (0.76/1.53/2.3 mA), a duration of 30 min, and an intensity increasing every 10 min. In Group DMSO + EA, DMSO (with a concentration of 0.2%, 50µL/acupoint) was injected at bilateral “ST 36” of the model rats one hour before electroacupuncture, while GW4869 (with a concentration of 3 mg/mL, 50µL/acupoint) was injected at bilateral ST 36 of the model rats one hour before electroacupuncture in Group GW4869 + EA. The paw withdrawal latency (PWL) was used as the therapeutic effect index. Results The PWL of rats in each group at Hour 24 after modeling was significantly lower than that before modeling, indicating that the models were successfully established. After the electroacupuncture treatment, the PWL of rats showed an increasing trend in all groups. The PWL of Group GW4869 + EA (6.74 ± 1.09)s was lower than that of Group EA (7.72 ± 1.54)s on Day 1 after injection, but the difference was not statistically significant (P > 0.05). The PWL values of Group GW4869 + EA (7.72 ± 0.70)s, (7.87 ± 0.58)s were significantly lower than those of Group EA (9.96 ± 0.94)s, (9.66 ± 0.96)s (both P Conclusion It was preliminarily found that acupuncture analgesic effect was significantly reduced after local exosomes of acupoints were blocked, indicating that local exosomes of acupoints may be involved in the initiation process of acupuncture effect.

BACKGROUND Heel sticks account for most blood tests performed in neonates without analgesia because topical local anesthetics are ineffective on heel glabrous skin. We investigated the antinociceptive effect of an alternative topical analgesic, a vapocoolant spray, on hind paw glabrous skin of rat pups. The spray was applied by two methods: method 1 for 4 s at a distance of 8 cm and method 2 for 10 s at a distance of 18 cm. METHODS The rat pups were randomized to either method 1 (n = 32) or method 2 (n = 31). Vapocoolant spray was applied to one hind paw randomly, and saline spray was applied to the contralateral paw. The paws were exposed to a hotplate test to measure withdrawal latency time before and 30 s after the spray applications. Additionally, rat pups were tested for tissue toxicity in method 1 (n = 20) and method 2 (n = 20) after application of the vapocoolant spray before heel sticks three times a day for two consecutive days. Analyses of spray and method effects on hotplate withdrawal latency time were determined by nonparametric Wilcoxon tests to assess paired difference between vapocoolant spray and saline spray and to compare difference in medians between the two methods. RESULTS Method 1 and method 2 vapocoolant spray applications significantly prolonged the withdrawal latency time compared with saline, a median difference of 0.6 s (IQR 0.1-1.2) for method 1 and 9.5 s (IQR 5.5-10.7) for method 2 (a 15-fold longer latency time with method 2). Method-2 produced significantly longer withdrawal latency time than method 1 with a difference in median time of 8.9 s (CI: 95% 7.3-10.4 s, P

Background: There is accumulating evidence showing that exercise therapy may play an active role in peripheral neuropathic pain (NP). However, there have been no meta-analysis to investigate the effects of exercise on NP induced by peripheral nerve injury in rat models. Methods: PubMed, EMBASE, and Web of Science were searched from inception to January 2019. A random-effect model was implemented to provide effect estimates for pain-related behavioral test outcome. Mean differences (MDs) with 95% confidence intervals (CIs) were calculated. Results: Fourteen studies were included. For the mechanical withdrawal threshold, rats in the exercised group exhibited significantly higher thresholds than those in the control group, with a MD of 0.91 (95% CI 0.11–1.71), 3.11 (95% CI 1.56–4.66), 3.48 (95% CI 2.70–4.26), 4.16 (95% CI 2.53–5.79), and 5.58 (95% CI 3.44–7.73) at 1, 2, 3, 4, and 5 weeks, respectively. Additionally, thermal withdrawal latency increased in the exercised group compared with the control group, with a MD of 2.48 (95% CI 0.59–4.38), 3.57 (95% CI 2.10–5.05), 3.92 (95% CI 2.82–5.03), and 2.84 (95% CI 1.29–4.39) at 1, 2, 3, and 4 weeks, respectively. Subgroup analyses were performed for pain models, exercise start point, exercise forms, and duration, which decreased heterogeneity to some extent. Conclusion: This meta-analysis indicated that exercise provoked an increase in mechanical withdrawal threshold and thermal withdrawal latency in animal NP models. Exercise therapy may be a promising non-pharmacologic therapy to prevent the development of NP. Further, preclinical studies focused on improving experiment design and reporting are still needed.

Introduction: Gout arthritis is an inflammatory disease characterized by severe acute pain. The goal of pharmacological gout arthritis treatments is to reduce pain, and thereby increase the patient’s quality of life. The Kv7/M channel activators retigabine and flupirtine show analgesic efficacy in animal models of osteoarthritic pain. We hypothesized that these drugs may also alleviate gout arthritis pain. Objective: To determine the effects of retigabine and flupirtine on pain behavior associated with monosodium urate (MSU)-induced gout arthritis. Methods: The gout arthritis model was established with an intra-articular injection of MSU into the right ankle joint, animals were treated with retigabine or flupirtine, and pain-related behaviors were assessed. Results: Retigabine and flupirtine significantly increased the mechanical threshold and prolonged the paw withdrawal latency in a rat model of gout arthritis pain in a dose-dependent manner. The antinociceptive effects of retigabine and flupirtine were fully antagonized by the Kv7/M channel blocker XE991. Conclusion: Retigabine and flupirtine showed antinociceptive effects for MSU-induced gout pain at different times during pain development.

Background Chronic constriction injury (CCI) is widely used as an animal neuropathic pain model. Neuropathic pain is considered to exist when withdrawal latency to thermal stimulation is decreased after inducing a CCI to the sciatic nerve. However, it is known that CCI leads to changes in skin temperature and that skin temperature can affect withdrawal latency. Aim of this study was to compare withdrawal latencies of constricted and contralateral hind limbs, to thermal stimulation, at the same artificially-induced skin temperatures. Methods Neuropathic pain was induced by four ligatures on the left sciatic nerve in adult male Wistar rats. Withdrawal latencies were measured from the 11th to 14th day after ligation, in different ambient temperatures, using the plantar test (Hargreaves method). By changing ambient we produced different hind limb skin temperatures. Results Our results show that (1) CCI cause an increase in skin temperature; (2) the withdrawal latency was inversely related to ambient and skin temperature in the same manner for both the ligated and contralateral hind limbs; and (3) withdrawal latencies did not differ significantly for the ligated and contralateral hind limbs when the temperature of the hind limbs was artificially made the same (i.e., by changing the ambient temperature). Conclusions Withdrawal latencies to thermal stimulation did not differ on ligated and contralateral hind limb after CCI to the sciatic nerve if the temperature of the hind limbs was artificially or mathematically made the same. This finding may have significant impact on the interpretation results of neuropathic pain research.

Our clinical studies have demonstrated that gemcabene, a small molecule in late-stage clinical development, lowers pro-inflammatory acute-phase protein, C-reactive protein (CRP). This observation was further confirmed in a cell-based study showing inhibition of cytokine-induced CRP production. Based on these observations, in the present study, we tested the hypothesis that gemcabene may possess anti-inflammatory activities in animal models of inflammatory disease. Efficacy of gemcabene was investigated in rat models of carrageenan-induced thermal hyperalgesia (CITH), monosodium iodoacetate (MIA)-induced osteoarthritis (OA), and IL-6/IL-6sR-induced inflammation. We also evaluated efficacy of gemcabene in collagen antibody-induced joint swelling and arthritis in BALB/c mice. In CITH rat model, gemcabene administration attenuated paw withdrawal latency (60% at 30 mg/kg/d and 97% at 100 mg/kg/d) and showed improvement in joint swelling (-50% at 30 mg/kg/d) in MIA model of OA. These findings were further corroborated by IL-6/IL-6sR knee injection model in rat, showing 63 and 71% reduction in hind paw weight distribution at 10 and 30 mg/kg/d doses, respectively. In mouse model of monoclonal antibody-induced arthritis, a dose-dependent attenuation of joint swelling was observed. These results demonstrate that the anti-inflammatory activity of gemcabene previously observed in cell-based and in clinical studies also occurred in animal models of inflammation-induced arthritis and hyperalgesia. Thus, in addition to hypolipidemic efficacy, the anti-inflammatory activity of gemcabene may have additional benefits to patients with elevated vascular inflammation.

The current study aims at investigating the downstream targets of spinal Annexin A10 in modulating neuropathic pain.Paw withdrawal latency and paw withdrawal threshold were measured to evaluate the pain-associated behaviour in rats. The expression of spinal Annexin A10, phosphorylated-extracellular regulated kinase 1/2 and extracellular regulated kinase were detected by western blotting. The level of tumour necrosis factor-α and interleukine-1β was tested by enzyme-linked immunosorbent assay (ELISA) kits.Chronic constrictive injury caused pain hypersensitivity in rats, along with increased expression of spinal Annexin A10, phosphorylated-extracellular regulated kinase 1/2, tumour necrosis factor-α and interleukine-1β in rats. Knockdown of spinal Annexin A10 suppressed the chronic constrictive injury-induced hyperalgesia, and inhibited the chronic constrictive injury-induced increased expression of phosphorylated-extracellular regulated kinase 1/2, tumour necrosis factor-α and interleukine-1β in the spinal cord. Inhibition of spinal extracellular regulated kinase activation decreased the release of tumour necrosis factor-α and interleukine-1β, but did not change the increased expression of Annexin A10 caused by chronic constrictive injury.Annexin A10 contributed to the development of neuropathic pain by activating spinal extracellular regulated kinase signalling and the subsequent release of tumour necrosis factor-α and interleukine-1β in the spinal cord.

The aim was to investigate the mechanisms behind sensory stimulation which can be used to desensitize CNS in patients with atypical orofacial pain. Earlier studies have shown that the kappa-receptor in the periaqueductal gray (PAG) is involved in sensory stimulation induced antinociception. A possible antinociceptive role for dynorphin B (DynB) in supraspinal regions was tested. The behavioral effect of sensory stimulation in conscious rats, by stroking the fur, was tested using the nociceptive test hotplate and the hindpaw withdrawal latency (HWL) was measured. In anesthetized rats sensory stimulation during different modalities, stroking or pinching was performed and the microdialysis technique was used to determine the extra cellular level of DynB in the ventrolateral PAG. To evaluate the antinociception after sensory stimulation DynB was microinjected into the PAG and the effect was measured with the HWL to heat. The results showed that sensory stimulation in conscious rats significantly increased the HWL as an antinociceptive effect. Innocuous sensory stimulation such as stroking the fore paw significantly elevated the DynB level in the PAG compared to internal control. After pinching a tendency to delayed release of DynB was seen and a possible discharge of the nerve terminals could be speculated upon. The blood pressure did significantly increase after pinching but not after stroking. An intra-PAG injection of DynB into the PAG increased the HWL to heat after 24 h compared to basal level of HWL and to saline treated animals. In conclusion, DynB is involved in the antinociception that is triggered by sensory stimulation.

The aim of this study was to observe the changes in mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) in a rat model of incisional pain with nicotine dependence and withdrawal. Twelve Wistar rats were randomly divided into a control and a withdrawal group, with 6 rats per group. In the control group, the rats were raised in normal conditions for 7 days without any treatment. A model of plantar incisional pain was established in the right lower extremity and changes in the plantar MWT and TWL of the healthy and operative sides were observed for 7 successive days. In the withdrawal group, the rats were raised in normal conditions and treated with a subcutaneous injection of pure nicotine (3 mg/kg), 3 times each day for 7 days. The model of plantar incisional pain in the right lower extremity was established, and changes in bilateral plantar MWT and TWL were observed for 7 days. The operative side plantar MWT and TWL in the withdrawal group were significantly lower than those in the control group on postoperative days 1–7, respectively (P

Background Adenosine is an endogenous neuromodulator in both the peripheral and central nervous systems. Adenosine inhibits pain signals by hyperpolarizing neuronal membrane. Methods To clarify the effects of adenosine on pain signals, we tested intrathecal adenosine injection in two neuropathic pains (spinal cord compression and chronic constriction of sciatic nerve) and postoperative pain (plantar incision). Results In all three kinds of pain models, significant shortening of withdrawal latencies to thermal stimulation were detected from 24 h to 1 week after the surgery. Significant improvements of pain sensation were observed in all three models after intrathecal injection of Cl-adenosine 24 h after surgery. At 72 h after surgery, intrathecal Cl-adenosine injection inhibited hyperalgesia in the two neuropathic pain models but not in the postoperative pain model. Adenosine A1R messenger RNA (mRNA) expression significantly decreased in the plantar incision model. Adenosine A1R protein levels also decreased compared with the other two models and normal control. Conclusions These results suggest that adenosine effectively inhibits pain signals in neuropathic pain but is less effective in postoperative pain because of the decrease in adenosine A1 receptors.

In the present study we examined whether the descending facilitation from the rostral ventromedial medulla (RVM) is required for the enhancement of formalin-evoked nocifensive behavior following repeated forced swim stress. Rats were subjected to forced or sham swim stress for 3days. Withdrawal latency to noxious thermal stimuli and mechanical withdrawal threshold to von Frey filaments did not change significantly in both groups at 24h after the last stress session. The forced swim stress showed significantly enhanced nocifensive behavior to the subcutaneous administration of formalin at 2days after the last stress session (1330.1+/-62.8s), compared to the sham swim (1076+/-102.4s, p0.05) and naive groups (825.9+/-83.2s, p0.01). The destruction of the RVM with ibotenic acid led to prevent the enhancement of formalin-evoked nocifensive behavior in the forced swim group. These findings suggest that the descending facilitation from the RVM may be involved in the enhancement of formalin-evoked nocifensive behavior following the forced swim stress.

Background Nitrous oxide (N(2)O) acts on supraspinal noradrenergic neurons to produce analgesia, but it is unclear if analgesia contributes to N(2)O's immobilizing effects. We tested the hypothesis that N(2)O minimum alveolar anesthetic concentration (MAC) is unchanged after selective ablation of supraspinal noradrenergic neurons, or in naive animals at N(2)O exposure timepoints when analgesia is absent. Methods We determined tailflick latency (TFL) and hindpaw withdrawal latency (HPL) under 70% N(2)O, N(2)O MAC, and isoflurane MAC before and after intracerebroventricular injections of anti-dopamine-beta hydroxylase conjugated to saporin (SAP-DBH; n = 7), or a control antibody conjugated to saporin (n = 5). In a separate group of naive rats (n = 8), N(2)O MAC was determined at 25-45 min after initiation of N(2)O exposure (during peak analgesia) and again at 120-140 min (after TFL and HPL returned to baseline). Results After 30 min of N(2)O exposure, TFL and HPL increased significantly but declined back to baseline within 120 min. N(2)O did not produce analgesia in rats that received SAP-DBH. However, N(2)O and isoflurane MAC were not significantly different between SAP-DBH and control-injected animals (Mean +/- sd for N(2)O: 1.7 +/- 0.1 atm vs 1.7 +/- 0.2 atm; isofurane: 1.6 +/- 0.2% vs 1.7 +/- 0.2%). In naive animals, N(2)O MAC was not different at the 30 min period compared with the 120 min period (1.8 +/- 0.1 atm vs 1.8 +/- 0.2 atm). Conclusions Destroying brainstem noradrenergic neurons or prolonged exposure to N(2)O removes its analgesic effects, but does not change MAC. The immobilizing mechanism of N(2)O is independent from its analgesic effects.

The paw withdrawal latency to thermal radiant heat stimuli is a widely used nociceptive measure to study hyperalgesic mechanisms. In the present study, in addition to the paw withdrawal latency, two behavioral components of pain behaviors, paw licking and paw lifting have been characterized and quantified. The thermal stimuli were successively applied to the plantar surface of the rat hind paws and recorded the behavioral responses to each of the stimuli. Noxious radiant heat significantly increased the frequency of paw lifting behavior in naïve rats. On the other hand, the thermal stimuli significantly evoked the occurrence of paw licking behavior in inflamed paws and it was maintained at all time points of measurements. The paw withdrawal latency decreased in inflamed rats in comparison with control rats. These data informs that noxious radiant heat specifically evokes the frequency of paw lifting behavior in normal physiological condition, and paw licking behavior in a pathological inflammatory condition. These findings suggest that in addition to the measurement of PWL, scoring of paw licking and lifting behaviors will improve the sensitivity of this pain test.

Noxious stimulation at one site on the body can inhibit noxious stimulation at distal body sites. This has been extensively demonstrated for somatic stimuli, but less so for visceral stimuli. In the present study we present a model for visceral inflammatory stimuli inhibiting somatic thermal sensitivity in awake rats. Colonic inflammation induced by mustard oil increases the hindpaw and forepaw withdrawal latency from a noxious radiant heat source by 35-50% compared to baseline responses. The duration of the effect is dose-dependent. The withdrawal latency in control rats (mineral oil in colon, mustard oil on skin) was not affected. Rotarod performance was not affected by 5% mustard oil indicating that colonic inflammation did not produce a general malaise or decrease in motor performance. These data suggest that visceral inflammation in the rat decreases somatic sensitivity similar to that reported by patients with colonic hypersensitivity from irritable bowel syndrome or inflammatory bowel diseases.

The present study investigated the effect of intraperiaqueductal grey injection of nociceptin/orphanin FQ (N/OFQ) and an antagonist (Nphe(1))nociceptin(1-13)NH(2) on the hindpaw withdrawal response to thermal and mechanical stimulation in rats. N/OFQ (5 nmol) significantly decreased the nociceptive thresholds in both tests and 1, 5 and 10 nmol of (Nphe(1))nociceptin(1-13)NH(2) significantly reversed this effect in a dose dependent way. Our results demonstrate, that N/OFQ has a nociceptive action, possibly through inhibition of PAG neurons. This effect is blocked by the antagonist (Nphe(1))nociceptin(1-13)NH(2) probably via ORL1 receptors in the periaqueductal grey.

Background Previously, the authors developed and characterized a rat model for postoperative pain to learn more about pain produced by incisions. In this study, the responses to heat and mechanical stimuli were evaluated directly on or adjacent to the incision and at varying distances from the incision. Methods Rats were anesthetized with halothane and incisions were made at different locations in the plantar aspect of the foot. The response frequency to a blunt mechanical stimulus, the withdrawal threshold to von Frey filaments (15-522 mN), and the withdrawal latency to radiant heat were measured. Rats were tested before surgery, 2 h later, and then daily through postoperative day 9. Results After plantar incision, persistent hyperalgesia was observed immediately adjacent to or directly on the incision to punctate and blunt mechanical stimuli, respectively. The withdrawal threshold to punctate stimuli applied 1 cm from the incision was decreased through postoperative day 1. In a transitional area, between the distant and adjacent sites, the withdrawal threshold was intermediate and the duration of hyperalgesia was transient. Heat hyperalgesia was persistent but present when the stimulus was applied to the site of injury but not to a distant site. Conclusion Robust primary hyperalgesia to punctate and blunt mechanical stimuli was present. Hyperalgesia distant to the wound, or secondary hyperalgesia, occurred in response to punctate mechanical stimuli, was short-lived, and required greater forces. These results suggest that the most persistent pain behaviors in this model are largely primary hyperalgesia.

Repeated oxytocin injections cause lowered blood pressure, decreased cortisol levels, increased withdrawal latency, increased release of vagally controlled gastrointestinal hormones, and increased weight gain. Together, these effects form an antistress pattern. Nonnoxious sensory stimuli release oxytocin and induce an effect spectrum similar to the one caused by oxytocin injections.

Ropivacaine has been associated with transient heat hyperalgesia in sciatic nerve blocks in rat. The goal of the present study was to evaluate the hypothesized presence of transient heat hyperalgesia after perineural injection of ropivacaine with a secondary subanalysis of 2 published studies. Paw withdrawal latency was used to assess the duration of sensory blockade and presence of heat hyperalgesia at 210, 240, 270, and 300 minutes and 24 hours after injection. The analysis revealed hyperalgesia at a single time point (240 minutes after injection; mean difference, −0.60 seconds; P = 0.012) that resolved within 30 minutes, and there was no other significant hyperalgesia at other time points. Although statistically significant, the single time point measurement represented only an 11% change from baseline and was no longer present 30 minutes later. These data support the need for a reevaluation of the interpretation that pain can be worsened by perineural ropivacaine injection.

Intrathecal delivery of drugs is an important method in pain research in order to investigate pain-relevant effects in the spinal cord in vivo. Here, we describe a method of intrathecal drug delivery by direct lumbar puncture in mice. The procedure does not require surgery, is rapidly performed, and does not produce neurological deficits. If cGMP analogs are injected, a state of transient hindpaw hypersensitivity can be induced which is quantifiable by measurement of hindpaw withdrawal latency in response to mechanical stimulation.

In a rat model of unilateral hindpaw inflammation, it is unclear whether the coeruleospinal modulation system is active at spinal segments distant from the inflamed plantar region, such as the cervical segments. To clarify this query, in the present study we measured paw withdrawal latency (PWL) to thermal stimuli on four paws (both forepaws and both hindpaws) following induction of inflammation and compared PWLs between rats with bilateral lesions of the locus coeruleus/subcoeruleus (LC/SC) and rats with sham operation. Unilateral hindpaw inflammation was produced by a subcutaneous injection of carrageenan (2 mg in 0.15 ml saline). Prior to carrageenan injection, in all four paws, PWLs did not differ between the LC/SC-lesioned and the sham-operated rats. Four hours after carrageenan injection, PWLs in the inflamed left hindpaw decreased significantly in both the LC/SC-lesioned and the sham-operated rats. The decreased PWLs of the LC/SC-lesioned group were significantly shorter than those of the sham-operated group. These phenomena which were observed in the inflamed left hindpaw were also observed in the non-inflamed left forepaws. In the right forepaws and the right hindpaws, no significant change in PWL was observed between before and 4 h after injection in both the sham-operated and the LC/SC-lesioned rats. These results suggest that unilateral hindpaw inflammation activates the coeruleospinal modulation system and that this modulation system is active not only at the lumbar segments but also at the cervical level where spinal segments are distant from the inflamed plantar region.

The purpose of the present study was to establish whether supraspinal structures modulate mechanical `adjacent hyperalgesia'. After a chronic sciatic cut, the paw withdrawal threshold to mechanical stimulation was lower, and the latency of noxious radiant heat-induced withdrawal reflex was shorter at the traumatized side than at the intact side. Then the rats were spinalized, and the withdrawal threshold to mechanical stimulus increased at the injured side, but the withdrawal latency induced by noxious heat decreased at the intact side. No side differences between the injured and the intact side could be detected after spinalization. Thus supraspinal structures may participate in maintenance of mechanically evoked paw withdrawal reflex after a sciatic injury.

We have examined the stability and sources of variation within the nociceptive model of rat hind paw withdrawal from an under-glass radiant stimulus (Hargreaves et al., 1988) using a system where stimulus intensity and floor temperature can be controlled and reproducibly changed. The current study demonstrates that: (i) increased stimulus intensity with a fixed surface temperature is associated with a monotonic decrease in mean response latency and its variance; (ii) for a fixed stimulus intensity, the mean paw withdrawal latency and variance increased as the glass floor temperature is lowered from 30 degrees C to room temperature (25 degrees C). Using subcutaneously-implanted thermocouples and a 30 degrees C glass surface, the subcutaneous paw temperature observed at an interval corresponding to the time at which the animal displayed a paw withdrawal did not differ across multiple heating rates (41-42.5 degrees C). This finding is in agreement with human studies of pain thresholds and C-fiber activity. These studies emphasize the importance of maintaining a fixed surface temperature to reduce experimental variability and the utility of this apparatus across multiple stimulus intensities to define agonist efficacy.

We recently demonstrated an olfactorily induced tail skin temperature drop in saline-injected rats exposed to an oxytocin-injected cagemate, an effect abolished by olfactory impairment. Treatment with oxytocin may induce both nociceptive and anti-nociceptive effects. The contrasting effects likely depend on the model and dosage used. Here we report an increased hindpaw withdrawal latency in response to nociceptive heat following the subcutaneous administration of oxytocin (1 mg/kg). An increased withdrawal response latency was also found in the untreated cagemates of an oxytocin-treated rat. The anti-nociceptive effect was abolished in oxytocin-antagonist-injected cagemates. Our results suggests that an olfactorily induced oxytocinergic mechanism is activated in the cagemates of an oxytocin-injected rat promoting anti-nociception.

The elevated T-maze test of anxiety has been used to separate in the same rat conditioned from unconditioned responses of fear/anxiety. The test apparatus consists of three elevated arms—one enclosed and two open. Inhibitory avoidance—representing learned fear—is measured by recording the time taken to leave the enclosed arm in three consecutive trials. Unconditioned fear is evaluated by recording the time to escape from the open arm. In this study we investigated procedural questions raised by the use of the elevated T-maze. Experiment 1 showed that restraining the animals at the end of the enclosed arm for 30 s did not change the first (baseline) latency to leave this arm, indicating that aversion for the hands of the experimenter is not a key motivation for this response. The results of Experiment 2 indicated that open-arm experience, but not handling stress is the main cause for inhibitory avoidance acquisition, because rats trained in a T-maze with the three arms enclosed did not show the usual increase in withdrawal latency along the three consecutive trials. The same experiment also showed that the latency to leave the open arm did not undergo habituation over five consecutive trials, evidencing an aversive motivation for this response. The importance of open-arm experience for inhibitory avoidance acquisition was further suggested by the results of Experiment 3, as the removal of a shield that prevents perception of openness tended to increase avoidance latency in the elevated T-maze from the first trial.

Background: Because genetic manipulation is commonly accomplished in mice, mouse models for pain haveadvanced our understanding of the mechanisms of persistent pain. The purpose of this experimental study isto develop a mouse model for understanding incision induced postoperative pain.Methods: A longitudinal incision was made at the hindpaw of male DBA/2 mice. The withdrawal frequency(WF) from applications of von Frey filaments and the response frequency (RF) to blunt mechanical stimulationwere examined in an incision group and a control grouP. The withdrawal latency (WL) to radiant heat and apain score based on weight bearing were also measured. Tests were performed 1 day before incision, and2 hours, 1-3 days, 5 days and 7 days after incision.Results: The WF for the strongest filament was 35.0 t 9.1% before incision and this increased to 100.0i: 0% at 2 hours and to 65.0 + 9.1% at 7 days after incision. The RF to the blunt stimulus was 4.1 d: 4.1%before incision and 100.0 + 0.0% at 2 hours and 42.8 + 10.8% at 7 days after incision. The WL was 6.6+ 0.5 sec before incision and 2.4 + 0.3 sec at 2 hours and 5.9 + 0.6 sec at 7 days after incision. Thepain score increased from 1.1 d: 0.8 to 7.4 d: 1.5 at 2 days after incision.Conclusions: A mouse model of acute postoperative pain was developing by making a surgical incision inthe mouse hindpaw. Mechanical hyperalgesia and allodynia lasting for several days demonstrate that this modelhas similarities to the human post-operative pain state. Future studies will allow us to further investigate thegenetic and molecular mechanisms of incisional pain. (Korean J Pain 2008; 21: 18-26)

Background: It has been known that melatonin is involved in the modulation of nociceptive transmission. However, the effect of melatonin administered spinally has not been examined. Therefore, we examined the effect of melatonin on the formalin-induced or thermal-induced nociception at the spinal level. Methods: Intrathecal catheter was inserted into the subarachnoid space of male Sprague-Dawley rats. Pain was assessed by formalin test (induced by injection of 50μl of a 5% formalin solution to the hindpaw) or Hot-Box test (induced by radiant heat application to the hindpaw). The effect of intrathecal melatonin was examined on flinching behavior in the formalin test or withdrawal response in Hot-Box test. Results: Intrathecal melatonin produced a limited, but dose-dependent reduction of the flinching response during phase 1 and 2 in the formalin test. In addition, melatonin delivered at evening also decreased the flinching response in both phases of the formalin test. Melatonin restrictively increased the withdrawal latency in Hot-Box test. Conclusions: These results suggest that melatonin is active against the formalinand thermal-induced nocicpetion at the spinal level, but the effect is limited. (Korean J Pain 2006; 19: 137-141)

Background: The purpose of this study was to characterize an animal model of persistent mechanical hyperalgesia induced by repeated intraplantar injection of low pH saline. Methods: Saline, at pHs of 4, 5, 7 and 7.4, was intraplantarily injected into the hind paw twice, 5 days apart. To quantify the hyperalgesia, the withdrawal threshold (WT) to mechanical stimuli was measured on the following schedule: before the first injection; 2 h and 4 h after the first injection; before the second injection and 4 h and weekly (6 weeks) after the second injection. The paw withdrawal latency to radiant heat was also measured with pH 4 saline. Mechanical hyperalgesia (MH) due to an intraplantar, subcutaneous injection was compared with that of a gastrocnemius muscle injection at pH 4. Results: Two unilateral injections of pH 4 and 5 saline solutions caused bilateral mechanical, but not heat hyperalgesia, which lasted 3 -4 weeks. An intraplantar injection of pH 4 saline showed a higher WT than the gastrocnemius muscle injection 4 hours after the second injection, but there was no significant difference between the two. Conclusions: Repeated intraplantar injections of low pH saline produced bilateral long-lasting MH in rats, but there was no significant difference in the WT between muscle and subcutaneous injections.

Background: Age-related differences in response to pain stimuli remain a controversial issue. The purpose of this study was to evaluate the effect of age on pain response using the formalin treated rats and streptozotin-induced diabetic rats tested for mechnical, cold allodynia and thermal hyperalgesia. Methods: We divided Sprague-Dawley rats into 3 groups by age, 5 weeks old (n = 10), 8 weeks old (n = 10), 12 weeks old (n = 10). Each group was divided into 2 subgroups (n = 5). One was formalin tested and the other was injected with streptozotin 75 mg intraperitoneally for succesive two days. On the 3rd day after injection, we examined mechnical allodynia using a von Frey filament, and tested thermal hyperalgesia using a tail immersion test in 50 or 5 water. Results: In the formalin test, pain response was higher in the younger age group (P in-induced diabetic rats, no difference in mechanical allodynia was observed between the subgroups. In the 50 water thermal hyperalgesia test, withdrawal latency decreased in each group after streptozotin injection (P water cold allodynia test, the 5-week animals showed a lower withdrawal latency than the other age groups (P

The saposin C-derived peptide TX14(A) prevents onset of functional and structural disorders in the peripheral nerve of diabetic rats. We have now investigated the ability of TX14(A) to alleviate behavioral indices of abnormal pain perception in adult female rats 4-6 weeks after onset of STZ-induced diabetes. Untreated diabetic rats exhibited tactile allodynia (response threshold = 3 ± 1 g) compared to age-matched controls (10 ± 1g). A single ip injection of TX14(A) transiently alleviated tactile allodynia, with an effect that was maximal 6 hours (11 ± 1g) after injection and diminished within 48 hours. Maximal efficacy was seen with a 1 mg/kg dose while no effects were noted in control rats. Control rats exhibited a transient thermal hyperalgesia (77 ± 5% of baseline paw withdrawal latency) 15 minutes after intrathecal delivery of substance P (30 nmol) that resolved within 30 minutes. Untreated diabetic rats exhibited substance P evoked thermal hyperalgesia of similar magnitude (82 ± 5% at 15 minutes) but of greater duration (83 ± 4% at 1 hour). Intrathecal delivery of TX14(A) 30 minutes before intrathecal substance P was without effect on the transient thermal hyperalgesia in control rats (74 ± 9% at 15 minutes). In diabetic rats, the prolonged thermal hyperalgesia was abolished by prior intrathecal delivery of TX14(A), although the transient thermal hyperalgesia (72 ± 8% at 15 minutes) remained. These studies show that TX14(A) can rapidly allevate diabetes-induced allodynia and hyperalgesia for up to 48 hours.

Latency to tail withdrawal from hot water was measured as a pain response before and after morphine injection in female rats. Morphine increased the withdrawal latency. Lesions in the preoptic forebrain region attenuated morphine analgesia.