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We found that morphine prevented LPS-induced TNF secretion in these cells. The observed inhibition was not due to morphine-induced TLR4 internalization and it was related to the blockage of preformed TNF secretion. LPS-induced TNF exocytosis in BMMCs was dependent on tetanus toxin\u2013insensitive vesicle-associated membrane proteins and calcium mobilization, as well as PI3K, MAPK, and I\u03baB kinase (IKK) activation. TNF secretion was also associated to the phosphorylation of synaptosomal-associated protein 23 (SNAP-23), which was found forming a complex with IKK in LPS-activated BMMCs. Morphine pretreatment prevented TLR4-dependent ERK and IKK phosphorylation. Analyzing the signaling events upstream of IKK activation, we found diminished TGF-\u03b2\u2013activated kinase 1 (TAK1) phosphorylation and TNFR-associated factor (TRAF) 6 ubiquitination in BMMCs pretreated with morphine and stimulated with LPS. Morphine pretreatment provoked a marked increase in the formation of a molecular complex composed of TRAF6 and \u03b2-arrestin-2. Naloxone and a combination of \u03bc and \u03b4 opioid receptor antagonists prevented morphine inhibitory actions. In conclusion, our results show that activation of \u03bc and \u03b4 opioid receptors with morphine suppresses TLR4-induced TNF release in mast cells, preventing the IKK-dependent phosphorylation of SNAP-23, which is necessary for TNF exocytosis, and this inhibition correlates with the formation of a \u03b2-arrestin-2\/TRAF6 complex. To our knowledge, these findings constitute the first evidence of molecular crosstalk between opioid receptors and the TLR4 signal transduction system in mast cells.<\/jats:p>","DOI":"10.4049\/jimmunol.1202658","type":"journal-article","created":{"date-parts":[[2013,8,20]],"date-time":"2013-08-20T01:29:04Z","timestamp":1376962144000},"page":"3400-3409","source":"Crossref","is-referenced-by-count":57,"title":["Morphine Prevents Lipopolysaccharide-Induced TNF Secretion in Mast Cells Blocking I\u03baB Kinase Activation and SNAP-23 Phosphorylation: Correlation with the Formation of a \u03b2-Arrestin\/TRAF6 Complex"],"prefix":"10.1093","volume":"191","author":[{"given":"Iris K","family":"Madera-Salcedo","sequence":"first","affiliation":[{"name":"Departamento de Farmacobiologia, Centro de Investigaci\u00f3n y de Estudios Avanzados del Instituto Polit\u00e9cnico Nacional , 14330 Mexico City,","place":["Mexico"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia L","family":"Cruz","sequence":"additional","affiliation":[{"name":"Departamento de Farmacobiologia, Centro de Investigaci\u00f3n y de Estudios Avanzados del Instituto Polit\u00e9cnico Nacional , 14330 Mexico City,","place":["Mexico"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Gonzalez-Espinosa","sequence":"additional","affiliation":[{"name":"Departamento de Farmacobiologia, Centro de Investigaci\u00f3n y de Estudios Avanzados del Instituto Polit\u00e9cnico Nacional , 14330 Mexico City,","place":["Mexico"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2013,9,15]]},"reference":[{"key":"2025032405341747000_r1","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1128\/CMR.16.2.209-219.2003","article-title":"Microbial infections, immunomodulation, and drugs of abuse","volume":"16","author":"Friedman","year":"2003","journal-title":"Clin. 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