Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / nÃo hà / Paclitaxel (PCX) foi o 1 antineoplÃsico efetivo no tratamento de cÃnceres refratÃrios a quimioterapia convencional. Clinicamente, induz artralgias e mialgias de carÃter incapacitante que comprometem a qualidade de vida e limitam o tempo de tratamento antitumoral, acometendo atà 57% dos doentes. Apesar destas repercussÃes clÃnicas nada foi descrito atà o momento, visando elucidar o envolvimento de citocinas prÃ-inflamatÃrias, na gÃnese da atividade hipernociceptiva do PCX, jà que a droga compartilha com o LPS uma via de sinalizaÃÃo desencadeada por receptores Toll (TLR-4 e TLR-2) para geraÃÃo de genes que codificam TNF-α. Dados da literatura sugerem que ocorra um âcross-talkâ entre esses dois membros da famÃlia Toll e, que agonistas diferentes de TLR-2 e TLR-4 sÃo capazes de induzir a ativaÃÃo de NF-αB, AP1 e MAP kinase e a geraÃÃo de TNF-α, citocina chave na cascata de liberaÃÃo de mediadores inflamatÃrios finais que atuam diretamente no nociceptor. Dados do laboratÃrio registraram que o zymosan (ZY) intrarticular em joelhos de ratos produz uma periartrite caracterÃstica da hipernocicepÃÃo no teste de incapacitaÃÃo articular (IA), e que PCX (8mg/kg) amplificou essa resposta quando se injetou  da dose do ZY. Tal amplificaÃÃo foi inibida com o prÃ-tratamento com inibidores de citocina e de prostanÃides. Objetivo. Investigar a participaÃÃo do TLR-4 e TNF-α na gÃnese do efeito potencializador do PCX na artralgia experimental induzida por ZY. Material e MÃtodos. Ratos foram prÃ-tratados Sc com talidomida (TLD), pentoxifilina, dexametasona, indometacina ou celecoxib e estimulados com subdose de ZY (250μg/animal; i-art). ApÃs a 1 medida do tempo de suspensÃo de pata (TSP) no teste de IA, os animais receberam PCX (8mg/kg; ip). Numa segunda etapa os ratos receberam durante trÃs dias consecutivos o prÃ-tratamento com atorvastatina (3, 10, 30mg/kg/dia; VO). Os seguintes parÃmetros foram avaliados: modulaÃÃo da hipernocicepÃÃo no teste de incapacitaÃÃo articular, dosagem de citocinas em lavado de joelho de ratos (TNF-α, IL-1α, Il-6, KC e CINC) e imunohistoquÃmica para TNF-α, IL-1α e TLR-4 no tecido sinovial. Resultados. Ficou demonstrado que PCX (8mg/kg) potencializa a artralgia experimental induzida por ZY em ratos avaliada pelo aumento significativo do TSP (p<0,001) na 4Âh de artrite em relaÃÃo ao controle no teste de IA. Tal efeito foi inibido de maneira significativa pelo prÃ-tratamento com TLD (45mg/kg) e essa inibiÃÃo foi associada à reduÃÃo dos nÃveis de TNF-α produzido pelas cÃlulas do tecido sinovial no lavado articular e da marcaÃÃo imunohistoquÃmica para TNF-. Da mesma forma a inibiÃÃo dessa resposta amplificadora do PCX foi ratificado pelo prÃ-tratamento com atorvastatina nas trÃs doses utilizadas no modelo, tambÃm sendo associado à diminuiÃÃo significativa dos nÃveis de TNF- no lavado articular e visÃvel reduÃÃo na marcaÃÃo imunohistoquÃmica para TNF-, IL-1 e TLR-4, nas trÃs doses utilizadas. ConclusÃes. PCX potencializa a hipernocicepÃÃo induzida por ZY por um mecanismo indireto sobre cÃlulas residentes da membrana sinovial que liberam TNF- provavelmente pela ativaÃÃo da NF-B via TLR-4/MD2, pois esse efeito potencializador foi inibido pela atorvastatina, um provÃvel antagonista de TLR-4. O TNF- liberado age iniciando a cascata de mediadores envolvidos com a dor inflamatÃria, o que justifica em parte as artralgias dos pacientes em tratamento com PCX. / Paclitaxel (PCX) was the first effective antineoplastic medicine in the treatment of tumors that do not respond to conventional chemotherapy. Clinically, it induces incapacitating arthralgias and myalgias that interfere with the patient quality of life and limit the duration of the treatment. This is observed in up to 57% of the patients using the drug. Despite these clinical manifestations, nothing has been published that could explain the involvement of pro-inflammatory cytokines in the triggering of the hypernociceptive effect of PCX, even though it is known that the drug shares with LPS a signaling pathway started by Toll-like receptors (TLR-2 and TLR-4) that activates genes coding for TNF-α. The literature suggests that there is a crosstalk between these two members of the Toll family and that different agonists of TLR-2 and TLR-4 are able to induce the activation of NF-kB, AP1 and MAP kinase in the generation of TNF-α, a key cytokines in the cascade liberating the final inflammatory mediators that act directly on the nociceptor. Data obtained in laboratory show that the injection of zymozan into rat knee-joints produces a periarthritis characteristic of the hypernociception seen in the knee joint incapacitation test and that PCX (8mg/kg) amplified the response when  of the zymozan (ZY) doses was injected. The amplification was inhibited when animals were pre-treated with inhibitors of cytokines and prostanoids. Objective: To study the role of TNF-α and TLR-4 on the initiation of the potentiating effect of PCX on the experimental arthralgia induced by ZY. Material and Methods: Rats were pre-treated Sc with thalidomide, pentoxifiline, dexametazone, indometacin and celecoxib and then stimulated with an intra-articular subdoses of ZY (250μg/animal). After the first measurement of the paw elevation time in the knee joint incapacitation test, the animals were treated with PCX (8mg/kg ip). On a second trial, rats were treated for three consecutive days with atorvastatin (3, 10, 30mg/kg/day; VO). The following parameters were evaluated: modulation of the effect on the knee joint incapacitation test (JIT), amount of cytokines in the ratâs knee lavage (TNF-α, IL-1 β, IL-6, KC and CINC) and immunohistochemistry for TNF-α, IL-1β and TLR-4 on synovial tissue. Results: It was shown that PCX (8mg/kg) potentiates the experimental arthralgia induced by ZY in the rats as evaluated by the significant increase in paw elevation time (p<0.001) at the 4th h of arthritis in relations to controls. Such effect was significantly inhibited by pre-treatment with thalidomide (45mg/kg) and the inhibition was associated with a decrease in the amount of TNF-α produced by synovial tissue cells and detected in the joint lavage and in the immunohistochemistry for TNF-α. Likewise the inhibition of the amplifying response to PCX was seen with pre-treatment with atorvastatin at the three doses used in the experiment, which was also associated with a lower TNF-α in the joint lavage and perceptible decrease in the immunohistochemistry for TNF-α, IL-1β and TLR-4. Conclusions: PCX potentiates the hypernociception induced by ZY through an indirect effect on synovial membrane resident cells that release TNF-α probably through activation of the NF-kB pathway by TLR-4/MD2, since the potentiating effect was inhibited by atorvastatin, a TLR-4 antagonist. Released TNF-α act starting the cascade of mediators involved in the inflammatory pain and this partially explains the arthralgia in patients treated with PCX.
| Identifer | oai:union.ndltd.org:IBICT/oai:www.teses.ufc.br:2737 |
| Date | 07 January 2009 |
| Creators | Mirlane GuimarÃes de Melo Cardoso |
| Contributors | Ronaldo de Albuquerque Ribeiro, Mariana Lima Vale, Francisco Airton Castro da Rocha, Adriana Rolim Campos Barros, Fernando de Queiroz Cunha, Max Victor Carioca Freitas |
| Publisher | Universidade Federal do CearÃ, Programa de PÃs-GraduaÃÃo em Farmacologia, UFC, BR |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFC, instname:Universidade Federal do Ceará, instacron:UFC |
| Rights | info:eu-repo/semantics/openAccess |
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