title: Endogenous cannabinoid anandamide impairs cell growth and induces apoptosis in chondrocytes.
abstract: Endocannabinoids has been described to be involved in articular degenerative disease by modulating nociception and immune system. However, the role of the endocannabinoid anandamide on chondrocyte cell viability is still unclear. Therefore, we decided to study anandamide's effects on chondrocytes viability and to evaluate its interactions with the catabolic factor TNF (tumor necrosis factor). Chondrocyte vitality was evaluated by MTT assay. We investigated LDH release, chromatin condensation, cleavage of focal adhesion kinase (FAK), and caspases-3, 8, and 9 activation. c-MYC mRNA levels were determined by RT-PCR. We studied by Western blot the activation patterns of AKT, AMPK, ERK, p38, and JNK kinases. Finally, we evaluate the effect of anandamide in TNF-induced caspase-3 cleavage. Anandamide decreased chondrocyte vitality independently of its receptors. It induced AMPK activation without LDH release. Anandamide induced chromatin condensation, activation of caspase-3, 8, and 9, and FAK cleavage. Surprisingly, despite anandamide inhibited cell proliferation, it increased c-MYC expression. Moreover anandamide inhibited AKT activation, whilst it induced a sustained activation of ERK, JNK, and p38. Finally, anandamide synergized with TNF-α in the cleavage of caspase-3. In conclusion, our findings suggest that anandamide, alone or in combination with TNF-α, may be a potential destructive agent in cartilage.
authors.affiliation: SERGAS-IDIS, Servizo Galego de Saude and Instituto de Investigación Sanitaria de Santiago, The NEIRID (NeuroEndocrine Interactions in Rheumatology and Inflammatory Diseases) Laboratory, Santiago University Clinical Hospital, Research Laboratory 9, Santiago de Compostela, Spain.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Gómez
authors.forename: Javier
authors.initials: J
authors.lastname: Conde
authors.forename: Morena
authors.initials: M
authors.lastname: Scotece
authors.forename: Verónica
authors.initials: V
authors.lastname: López
authors.forename: Francisca
authors.initials: F
authors.lastname: Lago
authors.forename: Juan Jesús
authors.initials: JJ
authors.lastname: Gómez Reino
authors.forename: Oreste
authors.initials: O
authors.lastname: Gualillo
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001095:Arachidonic Acids / Q000494:pharmacology*; D002186:Cannabinoids / Q000494:pharmacology*; D020169:Caspases / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D019902:Chondrocytes / Q000187:drug effects / Q000473:pathology*; D002843:Chromatin / Q000187:drug effects; D063388:Endocannabinoids / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology; D051379:Mice; D023421:Models, Animal; D053284:Polyunsaturated Alkamides / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D001095:Arachidonic Acids; D002186:Cannabinoids; D002843:Chromatin; D063388:Endocannabinoids; D053284:Polyunsaturated Alkamides; D014409:Tumor Necrosis Factor-alpha; D051057:Proto-Oncogene Proteins c-akt; D055372:AMP-Activated Protein Kinases; D020169:Caspases; C078814:anandamide
keywords: TNF; apoptosis; cannabinoids; chondrocyte physiology; signal transduction