FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A lesÃo de reperfusÃo (LR) ocorre em diversos orgÃos e tecidos durante o restabelecimento do fluxo sangÃÃneo apÃs episÃdios de isquemia prolongada. Em diversas Ãreas clÃnicas esta lesÃo à temida pelas complicaÃÃes decorrentes da demora no restabelecimento do fluxo vascular. Nos transplantes de ÃrgÃos, nos retalhos microcirÃrgicos e nas cirurgias de revascularizaÃÃo miocÃrdica, a LR, dependendo de sua intensidade, pode prejudicar o benefÃcio destes procedimentos. Dentre as estratÃgias propostas para atenuar a LR, o prÃ-condicionamento isquÃmico (PCI) consiste na induÃÃo de pequenos perÃodos de isquemia seguidos de reperfusÃo, realizados antes da isquemia prolongada. Este estudo foi avaliar o efeito do PCI da pata posterior na sobrevida de retalhos cutÃneos randomizados em dorso (RCRD) e o efeito antiinflamatÃrio local e sistÃmico deste fenÃmeno. Para tanto o PCI foi induzido atravÃs da isquemia da pata posterior de ratos Wistar por 10 minutos seguida de 30 minutos de reperfusÃo. Diferentes grupos de ratos foram submetidos à confecÃÃo de RCRD, à induÃÃo de edema de pata (EP), ipsilateral (IPS) e contralateral (CON), por carragenina (Cg) ou dextran (Dx), cistite hemorrÃgica (CH) por ifosfamida (IFO) ou lesÃo gÃstrica (LG) por indometacina (INDO). Os grupos controles (C) receberam o mesmo tratamento, porÃm sem PCI. A Ãrea de sobrevida (AS) do RCRD foi calculada pela superfÃcie de tecido viÃvel (cm2), o EP atravÃs da variaÃÃo do volume das patas (ml), o edema vesical (EV) pelo peso Ãmido da bexiga (mg), o aumento da permeabilidade vascular (APV) pelo extravasamento de azul de Evans (g), o Ãndice de lesÃo gÃstrica (ILG) pela soma das extensÃes das lesÃes encontradas na mucosa (mm), e a infiltraÃÃo neutrofÃlica (IN) pela atividade da mieloperoxidade (MPO) (U/mg). O PCI foi capaz de aumentar a AS do RCRD (C=4,16Â0,29 e PCI=6,42Â0,14, p<0,01). O EP e o APV induzidos por Cg (100g, sc) ou Dx (200g, sc), tanto na pata IPS quanto CON, foram reduzidos pelo PCI (EP IPS Cg â C=0,63+0,05 e PCI=0,26+0,02; APV IPS Cg â C=10,03+0,68 e PCI=5,82+1,18; EP IPS Dx â C=1,08+0,11 e PCI=0,54+0,06; APV IPS Dx â C= 24,38+3,02 e PCI=13,51+1,70; EP CON Cg â C=0,63+0,10 e PCI=0,30+0,05; APV CON Cg â C=16,16+2,18 e PCI=9,19+1,12; EP CON Dx â C=0,95+0,10 e PCI=0,49+0,10; APV CON Dx â C=24,09+2,97 e PCI=13,15+1,62; p<0,01). O PCI inibiu o EV e o APV da CH induzida por IFO (200mg/kg, ip) (EV â C=206,6+10,95 e PCI=126+5,66; APV â C=38,55+2,19 e PCI=17,74+2,18; p<0,001). Tanto a LG quanto a IN induzidas por INDO (20mg/kg, vo) foram bloqueadas pelo PCI (ILG â C=20,54+2,05 e PCI=2,00+0,93; IN â C=9,67+2,05 e PCI=3,49+0,83; p<0,001). Os resultados indicam que o PCI aumenta a sobrevida de RCRD e apresenta um efeito antiinflamatÃrio local e sistÃmico em modelos de inflamaÃÃo aguda. O esclarecimento do mecanismo atravÃs do qual o PCI atenua a lesÃo inflamatÃria poderà trazer importantes contribuiÃÃes em situaÃÃes clÃnicas onde esta lesÃo constitui um fator complicador, podendo propiciar o desenvolvimento de novas abordagens profilÃticas e/ou terapÃuticas. / The reperfusion injury (RI) occurs when there is delay in restoring blood flow to organs and tissues. Among the strategies proposed to attenuate the RI is the ischemic preconditioning (IPC), which consists of induction of brief ischemic periods followed by reperfusion performed before the sustained ischemic insult. In the first part, this study aimed to evaluate the role of nitric oxide (NO), cyclic guanosine monophosphate (cGMP) and ATP-sensitive potassium channels (K+ATP) in the inhibitory effect of hind limb IPC on the mice peritoneal cavity neutrophil migration (NM). In the second part, the objective was to evaluate the potential systemic antinociceptive effect of IPC in mechanical plantar hypernociception test (MPH; Von Frey) in rats and to evaluate the involvement of NO, cGMP and K+ATP channels in this event. In the first part, the IPC was induced by hind limb ischemia for 10 min followed by 30 min of reperfusion in wild and knockout mice to inducible NO synthase (iNOS -/-). The leukocyte rolling (LR), leukocyte adhesion (LA) and NM were induced by ip administration of Carrageenan (Cg, 500 g/cavity) and results expressed in number of leukocytes/min, number of adherent leukocytes/100 m2 cells and number of neutrophils x 106/cavity, respectively. Different groups of animals were treated with saline (SAL), Aminoguanidine (AG, sc, 100mg/kg), ODQ (ip, 8 mol/kg) or Glibenclamide (GBC, sc, 20 mg/kg) 30 min before IPC induction. Controls received the same treatment, but without IPC. In the second part, the IPC was induced by hind limb ischemia for 10 min followed by 30 min of reperfusion in male Wistar rats (180-200g). Cg (300 g, intraplantar) or Prostaglandin E2 (PGE2 - 400 ng, intraplantar) were used as hypernociceptive stimulus on left paw immediately after induction of IPC in contralateral paw. Different groups of animals were treated 30 min before induction of IPC with AG (100 g/kg, intraplantar), LNMMA (50 g, intraplantar), ODQ (8 g, intraplantar) or GBC (160 g, intraplantar). Controls received the same treatments, but without IPC. The quantification of MPH was performed through subtraction strength/pressure (g) required to cause withdrawal of paw in contact with an apparatus of eletronic von Frey mensuread before hypernociceptive stimulus, by measure obtained 3h after administration of Cg or PGE2. The LR, LA and NM induced by Cg in the peritoneal cavity were significantly (p <0,01) inhibited by hind limb IPC of wild animals (LR= 75.94%, LA= 56.51%, NM= 79.01 %), but these effects were not observed in animals iNOS -/-. The treatment of wild animals with AG and ODQ, but not with GBC, abrogated the inhibitory effect of IPC on NM induced by Cg. Treatment of animals with AG, GBC or ODQ did not significantly alter the NM induced by Cg in the peritoneal cavity. We also observed that IPC did not alter the TNF-, IL1- and CXCL1 chemokine levels induced by Cg in the peritoneal cavity. The mechanical hypernociception induced by Cg or PGE2 in left hind paw was significantly reduced (p <0,01) when IPC was performed in the right paw (55% and 68%). The treatment of animals with AG, LNMMA, ODQ or GBC, before IPC, abrogated the antinociceptive effect of IPC. Treatment of animals with AG, LNMMA, ODQ or GBC did not significantly alter the hypernociception induced by noxious stimulation. Our results show that IPC has an inhibitory effect on remote NM, with NO an important mediator involved in this process, probably through the cGMP pathway. This inhibitory effect of IPC does not depend on the opening of K+ATP channels, or inhibition of synthesis and/or release of pro-inflammatory cytokines. The elucidation of the mechanism by which IPC inhibits the NM may make important contributions to clinical situations where neutrophil infiltration is a complicating factor. The IPC also has a potent inhibitory effect on inflammatory pain, with NO seems to be one important mediator involved in this protective effect of IPC, acting through the cGMP/K+ATP channel pathway. This is the first demonstration described in the literature of systemic antinociceptive effect of IPC, and the elucidation of the mechanisms involved in this process is fundamental for pain management induced by several inflammatory stimuli.
| Identifer | oai:union.ndltd.org:IBICT/oai:www.teses.ufc.br:28 |
| Date | 22 June 2005 |
| Creators | Marcus VinÃcius Ponte de Sousa Filho |
| Contributors | Ronaldo de Albuquerque Ribeiro, AntÃnio Aldo Melo Filho, Roberto CÃsar Pereira Lima JÃnior, JosenÃlia Maria Alves Gomes |
| Publisher | Universidade Federal do CearÃ, Programa de PÃs-GraduaÃÃo em Cirurgia, 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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