Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/8506 |
Resumo: | Peripheral nerve injury disrupts the normal functions of neurons and leads to rapid and progressive alterations in structural skeletal muscle, such as muscle atrophy and fibrosis, causing functional deficits. Electrical stimulation (ES) has been recommended to treat denervated muscles. The best parameters of ES to minimize muscle atrophy due to denervation is controversial. Furthermore, it is not clear if ES can, in fact, affect reinnervation of denervated muscles. Thus, this thesis has two main objectives: 1) to verify if ES, applied to denervated muscles by surface electrodes, can affect neuromuscular recovery after nerve crush injury in rats; and 2) to assess the impact of ES on fibrosis establishment in denervated muscles. Two manuscripts were produced and used the same experimental groups. Thirtyfive Wistar rats were divided into 5 groups: (1) Normal (N); (2) 7-day denervation (D7d); (3) 7-day denervation and ES (DES7d); (4) 15-day denervation (D15d); and (5) 15-day denervation and ES (DES15d). Tibialis anterior (TA) muscle denervation was induced by crushing the sciatic nerve. The ES protocol to stimulate TA muscles consisted of: 200 contractions per day divided into 4 consecutive series of 50 contractions, with 10-minute rests between each set. The following parameters were used: exponential monophasic pulse; width time: 2x chronaxie; amplitude: motor level; time On: 3s and Off: 6s. The sciatic functional index was calculated. Muscle excitability was assessed considering the rheobasis, chonaxie and accommodation. Morphometric analyses, such as the muscle fiber cross-sectional area and percentage of connective tissue proliferation were used to characterize muscle morphology. Molecular markers related to reinnervation (neural cell adhesion molecule, NCAM), neuromuscular junction organization and maintenance (MuSK, Dok-7 and nicotinic Acetylcholine Receptors (nAChR) subunits), muscle mass control (atrogin-1, MuRF1, myoD and myostatin), fibrosis (TGF-β and myostatin), extracellular matrix remodeling (metaloproteinases, MMPs) and inflammation (TWEAK/Fn14) were investigated by molecular biology techniques such as western-blot, qPCR or zymography. The main results showed ES impaired natural recovery of denervated muscles accentuating disability, muscle atrophy and fibrosis, as well as reducing muscle excitability. These morphofunctional and electrophysiological changes were related to different modulations of all molecular markers investigated in a timely manner. Overall, this thesis provides evidence that ES can delay the reinnervation process by modulating factors related to NMJ stability and organization, as well as induced disability and muscle atrophy, and decreased muscle excitability. In addition, ES applied to denervated muscles induced muscle fibrosis by modulating inflammatory pathways and also extracellular matrix production and remodeling. Warnings should be given to rehabilitation teams when recommending ES to treat denervated muscles. |
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Pinheiro, Clara MariaRusso, Thiago Luiz dehttp://lattes.cnpq.br/2755480908047791http://lattes.cnpq.br/43601340122156376dc334ed-8fb1-47f6-be26-af4d19617ff82017-02-08T12:10:37Z2017-02-08T12:10:37Z2016-02-26PINHEIRO, Clara Maria. Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos. 2016. Tese (Doutorado em Fisioterapia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8506.https://repositorio.ufscar.br/handle/ufscar/8506Peripheral nerve injury disrupts the normal functions of neurons and leads to rapid and progressive alterations in structural skeletal muscle, such as muscle atrophy and fibrosis, causing functional deficits. Electrical stimulation (ES) has been recommended to treat denervated muscles. The best parameters of ES to minimize muscle atrophy due to denervation is controversial. Furthermore, it is not clear if ES can, in fact, affect reinnervation of denervated muscles. Thus, this thesis has two main objectives: 1) to verify if ES, applied to denervated muscles by surface electrodes, can affect neuromuscular recovery after nerve crush injury in rats; and 2) to assess the impact of ES on fibrosis establishment in denervated muscles. Two manuscripts were produced and used the same experimental groups. Thirtyfive Wistar rats were divided into 5 groups: (1) Normal (N); (2) 7-day denervation (D7d); (3) 7-day denervation and ES (DES7d); (4) 15-day denervation (D15d); and (5) 15-day denervation and ES (DES15d). Tibialis anterior (TA) muscle denervation was induced by crushing the sciatic nerve. The ES protocol to stimulate TA muscles consisted of: 200 contractions per day divided into 4 consecutive series of 50 contractions, with 10-minute rests between each set. The following parameters were used: exponential monophasic pulse; width time: 2x chronaxie; amplitude: motor level; time On: 3s and Off: 6s. The sciatic functional index was calculated. Muscle excitability was assessed considering the rheobasis, chonaxie and accommodation. Morphometric analyses, such as the muscle fiber cross-sectional area and percentage of connective tissue proliferation were used to characterize muscle morphology. Molecular markers related to reinnervation (neural cell adhesion molecule, NCAM), neuromuscular junction organization and maintenance (MuSK, Dok-7 and nicotinic Acetylcholine Receptors (nAChR) subunits), muscle mass control (atrogin-1, MuRF1, myoD and myostatin), fibrosis (TGF-β and myostatin), extracellular matrix remodeling (metaloproteinases, MMPs) and inflammation (TWEAK/Fn14) were investigated by molecular biology techniques such as western-blot, qPCR or zymography. The main results showed ES impaired natural recovery of denervated muscles accentuating disability, muscle atrophy and fibrosis, as well as reducing muscle excitability. These morphofunctional and electrophysiological changes were related to different modulations of all molecular markers investigated in a timely manner. Overall, this thesis provides evidence that ES can delay the reinnervation process by modulating factors related to NMJ stability and organization, as well as induced disability and muscle atrophy, and decreased muscle excitability. In addition, ES applied to denervated muscles induced muscle fibrosis by modulating inflammatory pathways and also extracellular matrix production and remodeling. Warnings should be given to rehabilitation teams when recommending ES to treat denervated muscles.A lesão nervosa periférica interrompe as funções normais dos neurônios e leva a alterações rápidas e progressivas no músculo esquelético, tais como a atrofia muscular e a fibrose, causando perdas funcionais. Para o tratamento dos músculos desnervados a estimulação elétrica (EE) tem sido utilizada. A escolha dos melhores parâmetros de EE para minimizar a atrofia muscular devido a desnervação, é controversa. Além disso, não está claro se a EE pode afetar, de fato, a reinervação de músculos desnervados. Assim, esta tese tem dois objetivos principais: 1) verificar se a EE, aplicada aos músculos desnervados com eletrodos de superfície, pode afetar a recuperação neuromuscular após axonotmese do nervo ciático em ratos; e 2) avaliar o impacto da EE no estabelecimento da fibrose do músculo desnervado. Dois manuscritos foram produzidos e contavam com os mesmos grupos experimentais. Trinta e cinco ratos Wistar foram divididos em 5 grupos: (1) Normal (N); (2) Desnervado 7 dias (D7d); (3) Desnervado e ES 7 (DES7d); (4) Desnervado 15 dias (D15d); e (5) Desnervado e ES 15 dias (DES15d). Foi realizada a axonotmese no nervo isquiático. O protocolo de EE do músculo tibial anterior consistiu em: 200 contrações por dia, divididas em 4 séries consecutivas de 50 contrações, com 10 minutos de descanso entre cada série. Foram utilizados os seguintes parâmetros: pulso monofásico exponencial; tempo: 2 vezes cronaxia; amplitude: nível motor; TON: 3s e TOFF: 6s. O índice funcional do ciático foi calculado. A excitabilidade muscular foi avaliada considerando a reobase, cronaxia e acomodação. Análises morfométricas, tais como área de secção transversa da fibra muscular e a porcentagem de proliferação do tecido conjuntivo foram utilizados para caracterizar a morfologia. Marcadores moleculares relacionados à reinervação como a N-CAM (molécula de adesão celular neural), organização e manutenção da junção neuromuscular (JNM) (MuSK, Dok-7 e receptores de acetilcolina), controle de massa muscular (atrogin-1, MuRF1, myoD e miostatina), fibrose (TGF-β e miostatina), remodelação da matriz extracelular (metaloproteinases) e, inflamação (TWEAK / Fn14) foram investigados por técnicas de biologia molecular como western-blot, qPCR ou zimografia. Os principais resultados mostraram que a EE provocou perda da recuperação natural dos músculos desnervados acentuando a perda funcional, a atrofia muscular e a fibrose, assim como, reduzindo a excitabilidade muscular. Estas alterações morfofuncionais e eletrofisiológicas foram relacionadas à diferentes modulações de todos os marcadores moleculares, no decorrer do tempo estudado. De modo geral, a presente tese forneceu provas de que a EE pode atrasar o processo de reinervação por fatores relacionados com a estabilidade e a organização da JNM, bem como a induzir à incapacidade e à atrofia muscular, com diminuição da excitabilidade muscular. Além disso, a EE aplicada aos músculos desnervados induziu fibrose através da modulação da via inflamatória e também pela produção e remodelamento da matriz extracelular. Cuidados devem ser tomadas pelas equipes de reabilitação ao utilizar a EE no tratamento de músculos desnervados.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Fisioterapia - PPGFtUFSCarEstimulação elétricaMetaloproteinaseFibrose e inflamaçãoElectrical stimulationMetalloproteinaseFibrosis e inflammationCIENCIAS DA SAUDE::FISIOTERAPIA E TERAPIA OCUPACIONALEfeito da estimulação elétrica sob a reinervação de músculos desnervados em ratosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisOnline60060069248a3f-f906-42f9-a3e2-f8bc31ca78afinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTeseCMP.pdfTeseCMP.pdfapplication/pdf2337703https://repositorio.ufscar.br/bitstream/ufscar/8506/1/TeseCMP.pdf27d55740b52010374ef6d09b5fb74efaMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstream/ufscar/8506/2/license.txtae0398b6f8b235e40ad82cba6c50031dMD52TEXTTeseCMP.pdf.txtTeseCMP.pdf.txtExtracted texttext/plain165262https://repositorio.ufscar.br/bitstream/ufscar/8506/3/TeseCMP.pdf.txt0f2b709a8d7badfcdf7626b60c392ec5MD53THUMBNAILTeseCMP.pdf.jpgTeseCMP.pdf.jpgIM Thumbnailimage/jpeg6450https://repositorio.ufscar.br/bitstream/ufscar/8506/4/TeseCMP.pdf.jpge881ceb0f0d7afd43adef462a8ecd249MD54ufscar/85062023-09-18 18:31:07.333oai:repositorio.ufscar.br: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Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:07Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos |
title |
Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos |
spellingShingle |
Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos Pinheiro, Clara Maria Estimulação elétrica Metaloproteinase Fibrose e inflamação Electrical stimulation Metalloproteinase Fibrosis e inflammation CIENCIAS DA SAUDE::FISIOTERAPIA E TERAPIA OCUPACIONAL |
title_short |
Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos |
title_full |
Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos |
title_fullStr |
Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos |
title_full_unstemmed |
Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos |
title_sort |
Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos |
author |
Pinheiro, Clara Maria |
author_facet |
Pinheiro, Clara Maria |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/4360134012215637 |
dc.contributor.author.fl_str_mv |
Pinheiro, Clara Maria |
dc.contributor.advisor1.fl_str_mv |
Russo, Thiago Luiz de |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/2755480908047791 |
dc.contributor.authorID.fl_str_mv |
6dc334ed-8fb1-47f6-be26-af4d19617ff8 |
contributor_str_mv |
Russo, Thiago Luiz de |
dc.subject.por.fl_str_mv |
Estimulação elétrica Metaloproteinase Fibrose e inflamação |
topic |
Estimulação elétrica Metaloproteinase Fibrose e inflamação Electrical stimulation Metalloproteinase Fibrosis e inflammation CIENCIAS DA SAUDE::FISIOTERAPIA E TERAPIA OCUPACIONAL |
dc.subject.eng.fl_str_mv |
Electrical stimulation Metalloproteinase Fibrosis e inflammation |
dc.subject.cnpq.fl_str_mv |
CIENCIAS DA SAUDE::FISIOTERAPIA E TERAPIA OCUPACIONAL |
description |
Peripheral nerve injury disrupts the normal functions of neurons and leads to rapid and progressive alterations in structural skeletal muscle, such as muscle atrophy and fibrosis, causing functional deficits. Electrical stimulation (ES) has been recommended to treat denervated muscles. The best parameters of ES to minimize muscle atrophy due to denervation is controversial. Furthermore, it is not clear if ES can, in fact, affect reinnervation of denervated muscles. Thus, this thesis has two main objectives: 1) to verify if ES, applied to denervated muscles by surface electrodes, can affect neuromuscular recovery after nerve crush injury in rats; and 2) to assess the impact of ES on fibrosis establishment in denervated muscles. Two manuscripts were produced and used the same experimental groups. Thirtyfive Wistar rats were divided into 5 groups: (1) Normal (N); (2) 7-day denervation (D7d); (3) 7-day denervation and ES (DES7d); (4) 15-day denervation (D15d); and (5) 15-day denervation and ES (DES15d). Tibialis anterior (TA) muscle denervation was induced by crushing the sciatic nerve. The ES protocol to stimulate TA muscles consisted of: 200 contractions per day divided into 4 consecutive series of 50 contractions, with 10-minute rests between each set. The following parameters were used: exponential monophasic pulse; width time: 2x chronaxie; amplitude: motor level; time On: 3s and Off: 6s. The sciatic functional index was calculated. Muscle excitability was assessed considering the rheobasis, chonaxie and accommodation. Morphometric analyses, such as the muscle fiber cross-sectional area and percentage of connective tissue proliferation were used to characterize muscle morphology. Molecular markers related to reinnervation (neural cell adhesion molecule, NCAM), neuromuscular junction organization and maintenance (MuSK, Dok-7 and nicotinic Acetylcholine Receptors (nAChR) subunits), muscle mass control (atrogin-1, MuRF1, myoD and myostatin), fibrosis (TGF-β and myostatin), extracellular matrix remodeling (metaloproteinases, MMPs) and inflammation (TWEAK/Fn14) were investigated by molecular biology techniques such as western-blot, qPCR or zymography. The main results showed ES impaired natural recovery of denervated muscles accentuating disability, muscle atrophy and fibrosis, as well as reducing muscle excitability. These morphofunctional and electrophysiological changes were related to different modulations of all molecular markers investigated in a timely manner. Overall, this thesis provides evidence that ES can delay the reinnervation process by modulating factors related to NMJ stability and organization, as well as induced disability and muscle atrophy, and decreased muscle excitability. In addition, ES applied to denervated muscles induced muscle fibrosis by modulating inflammatory pathways and also extracellular matrix production and remodeling. Warnings should be given to rehabilitation teams when recommending ES to treat denervated muscles. |
publishDate |
2016 |
dc.date.issued.fl_str_mv |
2016-02-26 |
dc.date.accessioned.fl_str_mv |
2017-02-08T12:10:37Z |
dc.date.available.fl_str_mv |
2017-02-08T12:10:37Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/doctoralThesis |
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doctoralThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
PINHEIRO, Clara Maria. Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos. 2016. Tese (Doutorado em Fisioterapia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8506. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/8506 |
identifier_str_mv |
PINHEIRO, Clara Maria. Efeito da estimulação elétrica sob a reinervação de músculos desnervados em ratos. 2016. Tese (Doutorado em Fisioterapia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8506. |
url |
https://repositorio.ufscar.br/handle/ufscar/8506 |
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por |
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openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de São Carlos Câmpus São Carlos |
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Programa de Pós-Graduação em Fisioterapia - PPGFt |
dc.publisher.initials.fl_str_mv |
UFSCar |
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Universidade Federal de São Carlos Câmpus São Carlos |
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