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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2021.tde-06082021-105430
Document
Author
Full name
Roberta Figueiroa de Souza
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2021
Supervisor
Committee
Castelucci, Patricia (President)
Kfoury Junior, José Roberto
Maifrino, Laura Beatriz Mesiano
Nogueira, Maria Ines
Title in Portuguese
Estudo do efeito do anti-TNFα nos neurônios entéricos e células gliaisentéricas de camundongos na colite ulcerativa experimental.
Keywords in Portuguese
Anti-TNF Adalimumabe
Célula Glial Entérica
Colite Ulcerativa
Neurônio Entérico
Abstract in Portuguese
A colite ulcerativa e a Doença de Crohn fazem parte das doenças inflamatórias intestinais (DII) e apresentam processos patofisiológicos como a necrose do intestino e dos neurônios entéricos e glias entéricas. As DII tem como o principal mediador inflamatório o fator de necrose tumoral alfa (TNFα). Este projeto tem como objetivo estudar os efeitos do anti-TNFα, o Adalimumabe (ADA), no plexo mioentérico na colite ulcerativa experimental. Para isto, os animais C57BL/6 receberam 3% de sulfato de sódio dextran (DSS) dissolvido em água potável por 7 dias até que todos os camundongos dos grupos administrados (grupos DSS e DSS+ADA) desenvolvessem inflamação intestinal equivalente à de humanos. O grupo Sham recebeu água pelo mesmo período. O grupo (DSS+ADA) recebeu o anti-TNFα no 2º dia de ingestão do DSS. O grupo ADA recebeu água por todo o período e injeção de anti-TNFα no 2º dia. Foi avaliado o código químico, o número de neurônios por gânglio e a área dos neurônios entéricos imunorreativos a óxido nítrico sintase neuronal (NOSn), colina acetil transferase (ChAT), pan neuronal PGP9.5 e o receptor TNFR2. Também foi feito dupla marcação do pan neuronal PGP9.5 com marcador glial (GFAP). Além disso, foi obtida a análise do perfil neuronal (αm²) dos neurônios NOSn-ir, ChAT-ir e PGP9.5 -ir. As análises qualitativas e quantitativas foram obtidas no microscópio de fluorescência Nikon 80i. Os resultados quantitativos mostram que a) o DSS foi um protocolo que induziu a colite ulcerativa, uma vez que os animais apresentaram alterações no peso com diarréia e sangramento; b) o receptor TNFR2 foi encontrado nos neurônios do plexo mioentérico nos grupos Sham, ADA, DSS e DSS+ADA; c) a colite ulcerativa experimental afetou os neurônios entéricos, pois houve diminuição dos neurônios imunorreativos a TNFR2, ChAT, NOSn e PGP9.5; d) a colite ulcerativa experimental afetou de maneira diferenciada as células gliais entéricas, com aumento nos grupos DSS e DSS+ADA; e) o grupo DSS+ADA dos neurônios NOSn-ir e ChAT-ir apresentaram recuperação do número de neurônios por gânglio; f) a colite ulcerativa experimental afetou o perfil neuronal com redução da área nos grupos DSS e DSS+ADA. Conclui-se que os neurônios do plexo mioentérico são imunorreativos ao receptor TNFR2, a inflamação pelo DSS causou alterações no plexo mioentérico e o tratamento com o anti-TNFα pareceu ser efetivo no tratamento da colite ulcerativa experimental.
Title in English
Effect of anti-TNFα on enteric neurons and enteric glial cells in experimental ulcerative colitis
Keywords in English
anti-TNF Adalimumab
Enteric Glial Cell
Enteric Neuron
Ulcerative Colitis
Abstract in English
Ulcerative colitis and Crohn's disease are part of inflammatory bowel diseases (IBD) and have pathophysiological processes such as bowel necrosis and enteric neurons and enteric glia. Also, the main inflammatory mediator is tumor necrosis factor alpha (TNFα). This project aims to study the effects of anti-TNFα, Adalimumab (ADA), on the myenteric plexus in experimental ulcerative colitis. For this, C57BL / 6 animals received 3% dextran sodium sulfate (DSS) dissolved in drinking water for 7 days until all mice in the administered groups (DSS and DSS + ADA groups) developed intestinal inflammation equivalent to humans. The Sham group received water for the same period. The group (DSS + ADA) received anti-TNFα on the 2nd day of DSS intake. The ADA group received water for the entire period and anti-TNFα injection on the 2nd day. The chemical code, the number of neurons per ganglion, and the area of the neuronal nitric oxide synthase (nNOS), choline acetyl transferase (ChAT), PGP9.5 neuronal pan and TNFR2 receptor immunoreactive enteric neurons were evaluated. Double labeling of PGP9.5 neuronal pan with glial marker (GFAP) was also performed. In addition, the neuronal profile analysis (m²) of nNOS-ir, ChAT-ir and PGP9.5-ir neurons was performed. Qualitative and quantitative analyzes were obtained using the Nikon 80i fluorescence microscope. Quantitative results show that a) DSS was a protocol that induced ulcerative colitis, since the animals presented weight changes with diarrhea and bleeding; b) TNFR2 receptor was found in the myenteric plexus neurons in the Sham, ADA, DSS and DSS + ADA groups; c) Experimental ulcerative colitis affected the enteric neurons, as there was a decrease in the TNFR2, ChAT, nNOS and PGP9.5 immunoreactive neurons; d) experimental ulcerative colitis affected differently the enteric glial cells, with increase in the DSS and DSS + ADA groups; e) the DSS + ADA group of nNOS-ir and ChAT-ir neurons showed recovery of neurons per ganglia; f) experimental ulcerative colitis affected the neuronal profile with reduced area in the DSS and DSS + ADA groups. It is concluded that myenteric plexus neurons are immunoreactive to the TNFR2 receptor, DSS inflammation caused changes in the myenteric plexus and anti-TNFα treatment appeared to be effective in the treatment of experimental ulcerative colitis.
 
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Release Date
2023-08-06
Publishing Date
2021-10-13
 
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