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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2019.tde-30072020-140957
Document
Author
Full name
Felipe Alexandre Machado
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2019
Supervisor
Committee
Castelucci, Patricia (President)
Assís Neto, Antônio Chaves de
Bittencourt, Jackson Cioni
Maifrino, Laura Beatriz Mesiano
Title in Portuguese
Papel do receptor P2X7 nos neurônios entéricos e nas células gliais entéricas na colite ulcerativa experimental.
Keywords in Portuguese
camundongo deficiente gene P2X7
Célula Glial Entérica
Colite Ulcerativa
Neurônio Entérico
Receptor P2X7
Abstract in Portuguese
O ATP é conhecido como um neurotransmissor e seus receptores são os da família P2X1-7. Foi demonstrada a presença de receptor purinérgico P2X2, 3, 7 no sistema nervoso entérico. A Colite Ulcerativa e Doença de Crohn apresentam processos patofisiológicos, a necrose do intestino e efeitos nos neurônios entéricos e glias entéricas. Tem sido demonstrada a participação do receptor P2X7 no processo inflamatório e necrose de células. Este projeto visa estudar o efeito da colite ulcerativa no plexo mioentérico e célula glial entérica do intestino grosso de animais deficientes para o receptor P2X7 (P2X7-/-, P2X7KO). Para isto, foi injetado 2, 4, 6, ácido trinitrobenzeno sulfônico (TNBS) no intestino grosso de animais C57BL/6 (WT) e deficientes do gene do receptor P2X7, no grupo sham foi injetado veículo. Foram coletados tecidos para análises 24h 4 dias após a administração. Foi usada técnica de duplas e triplas marcações de imunofluorescência para análises qualitativas e quantitativas que foram obtidas por microscópio de fluorescência Nikon 80i. Para a análise morfológica do intestino grosso foi usado o método de histologia convencional. Os resultados quantitativos demonstraram diminuição significante de 13,9% e 7,1% de células-ir ao receptor P2X7 nos grupos WT/Colite 24h e WT/Colite 4 dias, respectivamente. Não houve diminuição de neurônios-ir a NOS e ChAT do grupo KO/Colite de 4 dias. Não houve diminuição de neurônios-ir a PGP9.5 dos grupos KO/Colite de 24h e 4 dias. Houve diminuição de 19,3% de glias-ir a GFAP no grupo WT/Colite 24h e aumento de 19% no grupo WT/Colite 4 dias. Não houveram alterações morfológicas nos neurônios dos grupos de 24h. Houve aumento de 29,4% e 40,2% na área do perfil neuronal nos neurônios-ir a ChAT dos grupos WT/Colite 4 dias e KO/Colite 4 dias, respectivamente e aumento de 20,1% na área do perfil neuronal de neurônios-ir a PGP9.5 do grupo KO/Colite 4 dias. Análise histológica apresentou hiperemia e não úlceras, edema e infiltração celular nos grupos WT/Colite de 24h e 4 dias. Os grupos KO/Colite 24h e 4 dias foram menos afetados pela colite na análise histológica.
Title in English
Role of P2X7 receptor in enteric neurons and enteric glial cells in experimental ulcerative colitis.
Keywords in English
Enteric Glial Cell
Enteric Neuron
P2X7 gene deficient
P2X7 receptor
Ulcerative Colitis
Abstract in English
ATP is known as a neurotransmitter and its receptors are those of the P2X1-7 family. The presence of purinergic receptor P2X2, 3, 7 has been demonstrated in the enteric nervous system. Ulcerative Colitis and Crohn's Disease presents pathophysiological processes, necrosis and effects on enteric neurons and enteric glias. Has been demonstrated P2X7 receptor in the inflammatory process and cell necrosis. This project aims to study the effect of ulcerative colitis on the myenteric plexus and enteric glial cell of the large intestine of P2X7 receptor (P2X7-/-) deficient animals. For this, 2, 4, 6, trinitrobenzene sulfonic acid (TNBS) was injected into the large intestine of C57BL / 6 (WT) and P2X7 receptor gene deficient animals (P2X7-/-, P2X7KO). Tissues were collected for analysis 24h and 4 days after administration. A double and triple immunofluorescence labeling technique was used for qualitative and quantitative analyzes obtained by a Nikon 80i fluorescence microscope. For the morphological analysis of the large intestine the conventional histology method was used. Quantitative results showed a significant decrease of 13.9% and 7.1% of cells-ir to P2X7 receptor in the WT/Colitis 24h and WT/Colitis 4 days, respectively. There was no decrease in neurons-ir to NOS and ChAT of the 4 day KO/Colitis group. There was no decrease in neurons-ir to PGP9.5 of the 24h and 4 day KO/Colitis groups. There was a 19.3% decrease in GFAP glia-ir in the WT/Colitis 24h group and a 19% increase in the WT/Colitis 4 days group. There were no morphological changes in the neurons of the 24h groups. There was a 29.4% and 40.2% increase in neuronal profile area in the WT/Colitis 4 days groups and KO/Colitis 4 days group ChAT-ir neurons, respectively, and a 20.1% increase in neuronal profile area. of neurons-ir to PGP9.5 from KO/Colitis group 4 days. Histological analysis showed hyperemia and no ulcers, edema and cellular infiltration in the 24h and 4 days WT/Colitis groups. Groups KO/Colitis 24h and 4 days were less affected by colitis on histological analysis.
 
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Release Date
2022-07-30
Publishing Date
2021-10-13
 
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