u-0126 and Hypersensitivity

Opioid-induced hyperalgesia (OIH) is one of the major problems associated with use of opioids in perioperative and chronic pain management. The mechanism underlying this paradoxical phenomenon needs to be fully elucidated. Laterocapsular division of the central nucleus of amygdale (CeLC) has emerged as an important brain center for pain modulation, so we hypothesize that the activation of extracellular signal-regulated kinase (ERK) in CeLC may modulate OIH through strengthening synaptic transmission between neurons in the CeLC. Phospho-ERK in CeLC was first found to be increased significantly in OIH rats induced by repeated subcutaneous injection of fentanyl. Blockade of this fentanyl-induced ERK activation by microinjection of U0126, an ERK inhibitor, into the CeLC reversed the behavioral hypersensitivity in a dose-dependent manner. In vitro whole-cell recordings evaluating the change in synaptic transmission found that the frequency as well as amplitude of miniature excitatory postsynaptic currents recorded on CeLC neurons from OIH rats were fundamentally increased and were completely reversed by acutely applied U0126 (10 μM in the recording well). In vivo microinjection of U0126 into the CeLC reversed the spinal long-term potentiation in OIH rats. These results showed that fentanyl-induced hypersensitivity may occur partly through the mechanism of ERK activation and followed by the strengthening of synaptic transmission in CeLC neurons.. This study provides evidence that ERK in the laterocapsular division of the CeLC is a key contributor to the development of fentanyl-induced hypersensitivity. Targeting the superspinal central CeLC can inhibit spinal long-term potentiation and alleviate behavioral hyperreflexia induced by fentanyl.

Topics: Animals; beta-Glucosidase; Butadienes; Electric Stimulation; Enzyme Inhibitors; Excitatory Postsynaptic Potentials; Extracellular Signal-Regulated MAP Kinases; Fentanyl; Gene Expression Regulation; Hypersensitivity; In Vitro Techniques; Male; Narcotics; Nitriles; Pain Measurement; Pain Threshold; Physical Stimulation; Rats; Rats, Sprague-Dawley; Spinal Cord; Time Factors

Extracellular signal-regulated kinases 1/2 (ERK1/2) make important contributions to allergic responses via their regulation of degranulation, eicosanoid production, and cytokine expression by mast cells, yet the mechanisms underlying their positive effects on FcεRI-dependent signaling are not fully understood. Recently, we reported that mast cell activation and anaphylaxis are negatively regulated by AMP-activated protein kinase (AMPK). However, little is known about the relationship between ERK1/2-mediated positive and the AMPK-mediated negative regulation of FcεRI signaling in mast cells.. We investigated possible interactions between ERK1/2 and AMPK in the modulation of mast cell signaling and anaphylaxis.. Wild-type or AMPKα2(-/-) mice, or bone marrow-derived mast cells obtained from these mice, were treated with either chemical agents or small interfering RNAs that modulated the activity or expression of ERK1/2 or AMPK to evaluate the functional interplay between ERK1/2 and AMPK in FcεRI-dependent signaling.. The ERK1/2 pathway inhibitor U0126 and the AMPK activator 5-aminoimidazole-4-carboxamide-1-β-4-ribofuranoside similarly inhibited FcεRI-mediated mast cell signals in vitro and anaphylaxis in vivo. ERK1/2-specific small interfering RNA also mimicked this effect on FcεRI signals. Moreover, AMPKα2 knockdown or deficiency led to increased FcεRI-mediated mast cell activation and anaphylaxis that were insensitive to U0126 or activator 5-aminoimidazole-4-carboxamide-1-β-4-ribofuranoside, suggesting that the suppression of FcεRI signals by the inhibition of the ERK1/2 pathway relies largely on AMPK activation. ERK1/2 controlled AMPK activity by regulating its subcellular translocation.. ERK1/2 ablated the AMPK-dependent negative regulatory axis, thereby activating FcεRI signals in mast cells.

Topics: Aminoimidazole Carboxamide; AMP-Activated Protein Kinases; Anaphylaxis; Animals; Butadienes; Cell Degranulation; Cells, Cultured; Extracellular Signal-Regulated MAP Kinases; Hypersensitivity; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Nitriles; Receptors, IgG; Ribonucleosides; Signal Transduction

Extracellular signal‑regulated kinase (ERK) 1/2 in the spinal cord has been implicated in the development of neuropathic pain and inflammatory pain. However, a limited number of studies have investigated the role of spinal ERK in incisional pain. The present study aimed to determine the role of ERK in the spinal cord in incisional pain. Incisional pain was established in rats by a unilateral hind paw incision. ERK1/2 expression was analyzed by immunohistochemistry. Hypersensitivity to pain was evaluated by measuring the paw withdrawal threshold using the von Frey test. The mitogen‑activated protein kinase kinase (MEK) inhibitor, U0126, was administered 20 min prior to or 10 min following the incision by intrathecal or intraperitoneal injection. Phosphorylated ERK1/2 in the ipsilateral L4‑5 spinal superficial dorsal horn was activated 1 min following the incision, reached its peak level at 5 min and then returned to the basal level 20 min following the incision. Pretreatment, but not post‑treatment with U0126 markedly attenuated the pain hypersensitivity induced by the incision. Therefore, the present study indicates that the transient activation of spinal ERK1/2 contributes to the initiation of pain hypersensitivity following surgical incision.

Topics: Animals; Butadienes; Disease Models, Animal; Enzyme Activation; Hyperalgesia; Hypersensitivity; Male; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinase Kinases; Nitriles; Pain, Postoperative; Phosphorylation; Posterior Horn Cells; Rats; Rats, Sprague-Dawley; Skin; Spinal Cord

Honeybee's sting on human skin can induce ongoing pain, hyperalgesia and inflammation. Injection of bee venom (BV) into the intraplantar surface of the rat hindpaw induces an early onset of spontaneous pain followed by a lasting thermal and mechanical hypersensitivity in the affected paw. The underlying mechanisms of BV-induced thermal and mechanical hypersensitivity are, however, poorly understood. In the present study, we investigated the role of mitogen-activated protein kinase (MAPK) in the generation of BV-induced pain hypersensitivity.. We found that BV injection resulted in a quick activation of p38, predominantly in the L4/L5 spinal dorsal horn ipsilateral to the inflammation from 1 hr to 7 d post-injection. Phosphorylated p38 (p-p38) was expressed in both neurons and microglia, but not in astrocytes. Intrathecal administration of the p38 inhibitor, SB203580, prevented BV-induced thermal hypersensitivity from 1 hr to 3 d, but had no effect on mechanical hypersensitivity. Activated ERK1/2 was observed exclusively in neurons in the L4/L5 dorsal horn from 2 min to 1 d, peaking at 2 min after BV injection. Intrathecal administration of the MEK inhibitor, U0126, prevented both mechanical and thermal hypersensitivity from 1 hr to 2 d. p-ERK1/2 and p-p38 were expressed in neurons in distinct regions of the L4/L5 dorsal horn; p-ERK1/2 was mainly in lamina I, while p-p38 was mainly in lamina II of the dorsal horn.. The results indicate that differential activation of p38 and ERK1/2 in the dorsal horn may contribute to the generation and development of BV-induced pain hypersensitivity by different mechanisms.

Topics: Animals; Bee Venoms; Butadienes; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Hyperalgesia; Hypersensitivity; Imidazoles; Immunohistochemistry; Inflammation; Male; Mitogen-Activated Protein Kinase 3; Nitriles; p38 Mitogen-Activated Protein Kinases; Pain Threshold; Pyridines; Rats; Rats, Sprague-Dawley; Spinal Cord