Mechanosensitive Ion Channel TMEM63A Gangs Up with Local Macrophages to Modulate Chronic Post-amputation Pain / 神经科学通报·英文版

Post-amputation pain causes great suffering to amputees, but still no effective drugs are available due to its elusive mechanisms. Our previous clinical studies found that surgical removal or radiofrequency treatment of the neuroma at the axotomized nerve stump effectively relieves the phantom pain afflicting patients after amputation. This indicated an essential role of the residual nerve stump in the formation of chronic post-amputation pain (CPAP). However, the molecular mechanism by which the residual nerve stump or neuroma is involved and regulates CPAP is still a mystery. In this study, we found that nociceptors expressed the mechanosensitive ion channel TMEM63A and macrophages infiltrated into the dorsal root ganglion (DRG) neurons worked synergistically to promote CPAP. Histology and qRT-PCR showed that TMEM63A was mainly expressed in mechanical pain-producing non-peptidergic nociceptors in the DRG, and the expression of TMEM63A increased significantly both in the neuroma from amputated patients and the DRG in a mouse model of tibial nerve transfer (TNT). Behavioral tests showed that the mechanical, heat, and cold sensitivity were not affected in the Tmem63a-/- mice in the naïve state, suggesting the basal pain was not affected. In the inflammatory and post-amputation state, the mechanical allodynia but not the heat hyperalgesia or cold allodynia was significantly decreased in Tmem63a-/- mice. Further study showed that there was severe neuronal injury and macrophage infiltration in the DRG, tibial nerve, residual stump, and the neuroma-like structure of the TNT mouse model, Consistent with this, expression of the pro-inflammatory cytokines TNF-α, IL-6, and IL-1β all increased dramatically in the DRG. Interestingly, the deletion of Tmem63a significantly reduced the macrophage infiltration in the DRG but not in the tibial nerve stump. Furthermore, the ablation of macrophages significantly reduced both the expression of Tmem63a and the mechanical allodynia in the TNT mouse model, indicating an interaction between nociceptors and macrophages, and that these two factors gang up together to regulate the formation of CPAP. This provides a new insight into the mechanisms underlying CPAP and potential drug targets its treatment.

Referred Somatic Hyperalgesia Mediates Cardiac Regulation by the Activation of Sympathetic Nerves in a Rat Model of Myocardial Ischemia / 神经科学通报·英文版

Myocardial ischemia (MI) causes somatic referred pain and sympathetic hyperactivity, and the role of sensory inputs from referred areas in cardiac function and sympathetic hyperactivity remain unclear. Here, in a rat model, we showed that MI not only led to referred mechanical hypersensitivity on the forelimbs and upper back, but also elicited sympathetic sprouting in the skin of the referred area and C8-T6 dorsal root ganglia, and increased cardiac sympathetic tone, indicating sympathetic-sensory coupling. Moreover, intensifying referred hyperalgesic inputs with noxious mechanical, thermal, and electro-stimulation (ES) of the forearm augmented sympathetic hyperactivity and regulated cardiac function, whereas deafferentation of the left brachial plexus diminished sympathoexcitation. Intradermal injection of the α2 adrenoceptor (α2AR) antagonist yohimbine and agonist dexmedetomidine in the forearm attenuated the cardiac adjustment by ES. Overall, these findings suggest that sensory inputs from the referred pain area contribute to cardiac functional adjustment via peripheral α2AR-mediated sympathetic-sensory coupling.

Cutaneous allodynia is more frequent in chronic migraine, and its presence and severity seems to be more associated with the duration of the disease / A alodinia cutânea é mais frequente na migrânea crônica, mas sua presença e gravidade parecem estar associadas com a duração da doença

ABSTRACT Objective To evaluate cutaneous allodynia among patients with chronic and episodic migraine in a tertiary headache clinic. Methods 80 subjects with episodic migraine and 80 with chronic migraine were assessed in a tertiary hospital. The 12-item Allodynia Symptom Checklist/Brazil questionnaire was applied to classify subjects according to the presence and severity of cutaneous allodynia. Results Cutaneous allodynia was identified in 81.3% of the episodic migraine group and 92.5% of the chronic migraine group (p = 0.03). No increased association could be attributed to chronic migraine when adjusted by years with disease (PR = 1.12; 95%CI = 0.99 to 1.27; p = 0.06). The groups also did not differ in the severity of allodynia, and severe presentation was the most frequent. Discussion Both groups seemed to be similarly affected in the cephalic and extracephalic regions, with the same severity. Conclusion Cutaneous allodynia is more frequent in chronic migraine, and its presence and severity seems to be more associated with the duration of the disease.

RESUMO Objetivo Avaliar a característica da alodinia cutânea em indivíduos com migrânea crônica e episódica em um hospital terciário. Métodos 80 sujeitos com migrânea episódica e 80 com migrânea crônica de um hospital terciário foram avaliados. O questionário 12-item Allodynia Symptom Checklist/Brasil foi aplicado e classificou os sujeitos quanto a presença e severidade da alodinia cutânea. Resultados A alodinia cutânea esteve presente em 81,3% dos migranosos episódicos e 92,5% nos crônicos (p = 0.03). Nenhuma associação pode ser atribuída a migrânea ao ser ajustada pela variável anos com doença (PR = 1.12; 95%IC = 0.99 para 1.27; p = 0.06). Os grupos não diferiram em relação à severidade da alodinia e a classificação severa foi a mais frequente. Discussão Ambos os grupos pareceram ser igualmente afetados nas regiões cefálicas e extracefálicas com a mesma severidade. Conclusão A alodinia cutânea é mais frequente na migrânea crônica, mas a presença e severidade parece estar mais associada com a duração da doença.

Secondary hyperalgesia occurs regardless of unilateral or bilateral knee osteoarthritis involvement in individuals with mild or moderate level / A hiperalgesia secundária ocorre independentemente do envolvimento unilateral ou bilateral da osteoartrite de joelho em indivíduos com doença leve ou moderada

ABSTRACT Background: Secondary hyperalgesia in individuals with less severe levels of knee osteoarthritis remains unclear. The objective of this study was to measure the pressure pain threshold of individuals with mild or moderate knee osteoarthritis and compare with no osteoarthritis. Methods: Ten healthy controls and 30 individuals with mild or moderate knee osteoarthritis divided into two groups (unilateral and bilateral involvement) were included. Dermatomes in lumbar levels (L1, L2, L3, L4 and L5) and sacral level (S1 and S2), myotomes (vastus medialis, vastus lateralis, rectus femoris, adductor longus, tibialis anterior, peroneus longus, iliacus, quadratus lumborum, and popliteus muscles), and sclerotomes in lumbar levels (L1-L2, L2-L3, L3-L4, L4-L5 supraspinous ligaments), over the L5-S1 and S1-S2 sacral areas, pes anserinus bursae, and at the patellar tendon pressure pain threshold were assessed and compared between individuals with and without knee osteoarthritis. Results: Knee osteoarthritis groups (unilateral and bilateral) reported lower pressure pain threshold compared to the control group in most areas (dermatomes, myotomes, and sclerotomes). There were no between group differences in the supra-spinous ligaments and over the L5-S1 and S1-S2 sacral areas of the sclerotomes. No difference was seen between knee osteoarthritis. Conclusion: These findings suggest that individuals with mild to moderate knee osteoarthritis had primary and secondary hyperalgesia, independent of unilateral or bilateral involvement. These results suggest that the pain have to be an assertive focus in the clinical practice, independent of the level of severity or involvement of knee osteoarthritis.

RESUMO Introdução: A ocorrência de hiperalgesia secundária em indivíduos com níveis menos graves de osteoartrite de joelho ainda é incerta. O objetivo deste estudo foi medir o limiar de dor à pressão (LDP) de indivíduos com osteoartrite de joelho (OAJ) leve ou moderada e comparar com indivíduos sem osteoartrite. Métodos: Foram incluídos 10 controles saudáveis e 30 indivíduos com OAJ leve ou moderada, divididos em dois grupos (envolvimento unilateral e bilateral). Foi avaliado e comparado o LDP em dermátomos (L1, L2, L3, L4, L5, S1 e S2), miótomos (músculos vasto medial, vasto lateral, reto femoral, adutor longo, tibial anterior, fibular longo, ilíaco, quadrado lombar e poplíteo) e esclerótomos (ligamentos supraespinais L1-L2, L2-L3, L3-L4, L4-L5), sobre as áreas sacrais L5-S1 e S1-S2, bolsa anserina e tendão patelar entre os indivíduos com e sem OAJ. Resultados: Os grupos OAJ (unilateral e bilateral) relataram menor LDP em comparação com o grupo controle na maior parte das áreas (dermátomos, miótomos e esclerótomos). Não houve diferenças entre os grupos nos ligamentos supraespinais e ao longo das áreas sacrais L5-S1 e S1-S2 dos esclerótomos. Não foi observada qualquer diferença entre os indivíduos com OAJ. Conclusão: Esses achados sugerem que os indivíduos com OAJ leve a moderada tinham hiperalgesia primária e secundária, independentemente do acometimento unilateral ou bilateral. Esses resultados sugerem que a dor precisa ser um foco assertivo na prática clínica, independentemente do grau de gravidade ou envolvimento da OAJ.

Matrix metalloproteinase inhibitor COL-3 prevents the development of paclitaxel-induced hyperalgesia in mice

To study the potential of chemically modified tetracycline-3 [COL-3], a potent matrix metalloproteinase [MMP] inhibitor, to protect against the development of paclitaxel-induced painful neuropathy and its immunomodulatory effects. The reaction latency to thermal stimuli [hot plate test] of female BALB/c mice was recorded before and after treatment with paclitaxel [2 mg/kg i.p.], paclitaxel plus COL-3 [4, 20 or 40 mg/kg p.o.] or their vehicles for 5 consecutive days. Gene transcripts of CD11b [marker for microglia], 5 cytokines [IFN-gamma, IL-1 beta, IL-6, IL-10 and TNF- alpha] and 3 chemokines [CCL2, CXCL10 and CX3CL1] were quantified by real-time PCR in the brains, spinal cords and spleens of mice sacrificed on day 7 after treatment. Treatment with paclitaxel reduced the reaction latency time to thermal stimuli [thermal hyperalgesia] for 4 weeks, with maximum effect on days 7 and 10. The coadministration of paclitaxel with COL-3 40 mg/kg, but not lower doses, prevented the development of paclitaxel-induced thermal hyperalgesia. Treatment with paclitaxel alone or coadministration with COL-3 increased CD11b transcript levels in the brain but not in the spinal cord. Treatment with paclitaxel reduced IL-6 transcript levels in the spinal cord but did not alter the transcript levels of other cytokines or chemokines in the brain, spinal cord or spleen. The coadministration of COL-3 with paclitaxel significantly increased the transcript levels of IL-6 in the spleen and decreased CX3CL1 transcripts in the brain in comparison to treatment with paclitaxel alone. Our results indicate that the MMP inhibitor COL-3 protected against paclitaxel-induced thermal hyperalgesia and, thus, could be useful in the prevention of chemotherapy-induced painful neuropathy

Participation of the NO/cGMP/K+ATP pathway in the antinociception induced by Walker tumor bearing in rats

Implantation of Walker 256 tumor decreases acute systemic inflammation in rats. Inflammatory hyperalgesia is one of the most important events of acute inflammation. The L-arginine/NO/cGMP/K+ATP pathway has been proposed as the mechanism of peripheral antinociception mediated by several drugs and physical exercise. The objective of this study was to investigate a possible involvement of the NO/cGMP/K+ATP pathway in antinociception induced in Walker 256 tumor-bearing male Wistar rats (180-220 g). The groups consisted of 5-6 animals. Mechanical inflammatory hypernociception was evaluated using an electronic version of the von Frey test. Walker tumor (4th and 7th day post-implantation) reduced prostaglandin E2- (PGE2, 400 ng/paw; 50 µL; intraplantar injection) and carrageenan-induced hypernociception (500 µg/paw; 100 µL; intraplantar injection). Walker tumor-induced analgesia was reversed (99.3% for carrageenan and 77.2% for PGE2) by a selective inhibitor of nitric oxide synthase (L-NAME; 90 mg/kg, ip) and L-arginine (200 mg/kg, ip), which prevented (80% for carrageenan and 65% for PGE2) the effect of L-NAME. Treatment with the soluble guanylyl cyclase inhibitor ODQ (100% for carrageenan and 95% for PGE2; 8 µg/paw) and the ATP-sensitive K+ channel (KATP) blocker glibenclamide (87.5% for carrageenan and 100% for PGE2; 160 µg/paw) reversed the antinociceptive effect of tumor bearing in a statistically significant manner (P < 0.05). The present study confirmed an intrinsic peripheral antinociceptive effect of Walker tumor bearing in rats. This antinociceptive effect seemed to be mediated by activation of the NO/cGMP pathway followed by the opening of KATP channels.

Interaction of Morphine and Selective Serotonin Receptor Inhibitors in Rats Experiencing Inflammatory Pain

Citalopram and paroxetine are selective serotonin reuptake inhibitors and also have antinociceptive effects. We investigated the antiallodynic and antihyperalgesic effects of intrathecally administered morphine, citalopram, paroxetine, and combinations thereof, in a rat model in which peripheral inflammation was induced by complete Freund's adjuvant (CFA). Drugs were intrathecally administered via direct lumbar puncture. Mechanical allodynia was measured using a Dynamic Plantar Aesthesiometer. Thermal hyperalgesia and cold allodynia were determined by measuring latency of paw withdrawal in response to radiant heat and cold water. Behavioral tests were run before and 15, 30, 45, and 60 min after intrathecal injection. Intraplantar injection of CFA produced mechanical allodynia, thermal hyperalgesia, and cold allodynia. Intrathecally administered morphine (0.3 or 1 microg) had antiallodynic or antihyperalgesic effects (24.0%-71.9% elevation). The effects of morphine were significantly increased when a combination of citalopram (100 microg) and paroxetine (100 microg) was added (35.2%-95.1% elevation). This rise was reversed by naloxone and methysergide. The effects of citalopram and paroxetine were also reversed by naloxone and methysergide. We suggest that the mu opioid receptor and serotonin receptors play major roles in production of the antiallodynic and antihyperalgesic effects of morphine, citalopram, paroxetine, and combinations thereof, in animals experiencing inflammatory pain.

Ketamina, una antigua droga que renace ante el dolor: Parte II / Ketamine, an old drug coming back for pain management: Part II

Esta segunda parte trata el tema de la hiperalgesia, siguiendo el modelo de la hiperalgesia postoperatoria. Se sabe que a pesar del conocimiento de la neurofisiología del dolor, sigue teniendo una alta incidencia de dolor postoperatorio, dolor crónico postoperatorio y dolor crónico postraumático. En todas estas situaciones, la nocicepción del dolor está aumentada, dada principalmente por la presencia de hiperalgesia. Se sabe que su diagnóstico es difícil, pero es necesario sospecharla para tratarla precozmente y evitar las repercusiones negativas en el individuo. En la hiperalgesia existe activación de los receptores NMDA,se postula que al ser bloqueado por la ketamina, principal antagonista de estos receptores, se tendría menor hiperalgesia y, por ende, menor dolor.

This second part addresses hyperalgesia following the postoperative hyperalgesia model. It is well known that in spite of the wide knowledge regarding pain neurophysiology, it still has a great impact on postoperative pain, postoperative chronic pain and post traumatic chronic pain. In all those situations, pain nociception is increased mainly due to the presence of hyperalgesia. Diagnosing it is difficult, however, one should suspect it so to start treatment at an early stage and avoid negative consequences for patients. Hyperalgesia is present in NMDA receivers which, as some think, if blocked by Ketamine, the main antagonist of these receivers, hyperalgesia will be lessened, hence pain reduced.

Anti-nociceptive effect of duloxetine in mouse model of diabetic neuropathic pain.

The involvement of adenosinergic pathway in the anti-nociceptive effect of duloxetine, a balanced 5-HT/NE reuptake inhibitor, was evaluated in streptozotocin induced diabetic male albino mice of Laca strain. After four weeks of single injection of streptozotocin (200 mg/kg, ip), mice were tested in the tail immersion and hot-plate assays. Cerebral adenosine levels were measured by high-performance liquid chromatography (HPLC/PDA detector). Diabetic mice exhibited significant hyperalgesia along with increased plasma glucose, decreased body weights and reduced cerebral adenosine levels. Administration of duloxetine (5, 10 and 20 mg/kg, ip) to diabetic mice produced dose-dependent anti-nociceptive effect in both tail-immersion and hot-plate assays. Adenosine levels were also significantly and dose-dependently increased by different doses of duloxetine. The results demonstrated the involvement of adenosinergic pathway in duloxetine mediated anti-hyperalgesia in diabetic neuropathic pain.

Effect of the Combined Use of Tramadol and Milnacipran on Pain Threshold in an Animal Model of Fibromyalgia

BACKGROUND/AIMS: Acidic saline injections produce mechanical hyperresponsiveness in male Sprague-Dawley rats. We investigated the effect of milnacipran in conjunction with tramadol on the pain threshold in an acidic saline animal model of pain. METHODS: The left gastrocnemius muscle of 20 male rats was injected with 100 microL of saline at pH 4.0 under brief isoflurane anesthesia on days 0 and 5. Rats administered acidic saline injections were separated into four study subgroups. After determining the pre-drug pain threshold, rats were injected intraperitoneally with one of the following regimens; saline, milnacipran alone (60 mg/kg), milnacipran (40 mg/kg) plus tramadol (20 mg/kg), or milnacipran (40 mg/kg) plus tramadol (40 mg/kg). Paw withdrawal in response to pressure was measured at 30 min, 120 min, and 5 days after injection. Nociceptive thresholds, expressed in grams, were measured with a Dynamic Plantar Aesthesiometer (Ugo Basile, Italy) by applying increasing pressure to the right or left hind paw until the rat withdrew the paw. RESULTS: A potent antihyperalgesic effect was observed when tramadol and milnacipran were used in combination (injected paw, p=0.001; contralateral paw, p=0.012). This finding was observed only at 30 min after the combination treatment. CONCLUSIONS: We observed potentiation of the antihyperalgesic effect when milnacipran and tramadol were administered in combination in an animal model of fibromyalgia. Further research is required to determine the efficacy of various combination treatments in fibromyalgia in humans.

Barostato rectal en el síndrome de intestino irritable / Rectal barostat in irritable bowel syndrome

Background: Many patients with irritable bowel syndrome (IBS) have lowered sensory thresholds to rectal distention when compared to control subjects, a phenomenon called visceral hypersensitivity. Aim: To investigate the usefulness of a rectal barostat as a diagnostic tool in IBS and if there are differences in visceral hypersensitivity in different groups of IBS patients. Patients and Methods: Ten healthy subjects and 19 IBS patients, defined using Rome II criteria (12 with constipation, three with diarrhea and four alternating between diarrhea and constipation), were studied. Sequential isobaric rectal distentions, from 2 mmHg up to a maximal pressure of 52 mmHg or when the patients reported pain, were carried out. Visceral hypersensitivity was defined as a pain threshold under 38 mmHg. Results: Only 26 percent of IBS patients had visceral hypersensitivity (16 percent and 43 percent of patients with IBS and constipation and IBS and diarrhea or alternating symptoms, respectively, p =NS). Pain threshold in controls, patients with IBS and constipation and patients with IBS and diarrhea or alternating symptoms was 43.8±6.6, 45.3±9.2 and 40.8±9.2 mmHg, respectively (p =NS). Conclusions: Our results do not support the usefulness of the electronic rectal barostat as a diagnostic method to diagnose IBS.

Resveratrol, a polyphenolic phytoalexin attenuates thermal hyperalgesia and cold allodynia in STZ-induced diabetic rats.

The effects of resveratrol, a polyphenolic phytoalexin present in red wine have been investigated on hyperalgesia and cold allodynia in streptozotocin (STZ) induced diabetic rats. Diabetes was induced by a single intraperitoneal injection of streptozotocin (65mg/kg). After 4-weeks of STZ injection, diabetic rats exhibited a significant thermal hyperalgesia and cold allodynia along with increased plasma glucose and decreased body weights as compared with controls rats. Chronic treatment with resveratrol (10mg/kg orally) from week 4 to week 6 significantly attenuated the cold allodynia and thermal hyperalgesia. The results emphasize the role of oxidative stress in development of hyperalgesia and cold allodynia in diabetic animals and point towards the potential of resveratrol as an adjuvant therapy for the prevention and treatment of diabetic neuropathy.

A new mechanism of action of dipyrone: blockade of the release of a nociceptive factor from macrophages

Rat macrophage monolayers pre-treated with endotoxin release into the incubating fluid a factor (MW >10,000) capable of inducing writhing in mice (MNF). This release was inhibited by dipyrone (3.5-35 microng/ml) but not by indomethacin (0.5-2 microng/ml). Writhing in mice induced by the factor is blocked by dipyrone (0.5-50 mg/kg) and indomethacin (0.5-2 mg/kg). These results indicate that in addition to the previously described direct blockade of hyperalgesia by dipyrone, this drug may also affect the release of MNF, wich induces in vivo nociception through the release of prostaglandin-like substances