CGRP e SP em cultura de fibroblastos de queloide
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Arquivos
Data
2011
Autores
Tucci-Viegas, Vanina Monique [UNIFESP]
Orientadores
Hochman, Bernardo [UNIFESP]
Tipo
Tese de doutorado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Introdução: Sabendo-se que o estímulo neurogênico de origem
nociceptiva é considerado o deflagrador primário do processo de
cicatrização da pele, que o CGRP e SP aumentam a proliferação de
fibroblastos in vitro, e diante da falta de estudos correlacionando CGRP, SP
e p53 e apoptose em queloide torna-se importante investigar se os
neuropeptídeos da pele podem influenciar a expressão de p53 e a apoptose
no queloide. Uma possível associação entre neuropeptídeos e o gene p53
poderia representar um novo caminho para um melhor entendimento
fisiopatogênico das cicatrizes patológicas fibroproliferativas e, por
conseguinte, o advento de tratamentos mais específicos. Objetivo:
Investigar a expressão do gene p53 e o índice apoptótico em culturas de
fibroblastos de queloide, submetidos à ação dos neuropeptídeos CGRP e
SP. Métodos: Foram cultivados fibroblastos de queloide em meio de
cultura com adição de CGRP e SP, e controle. O índice apoptótico, e o
ciclo celular foram determinados por citometria de fluxo e a marcação de
proteínas-chave da apoptose por espectrofotometria. A atividade celular foi
avaliada ao longo do tempo e em períodos determinados, por MTT. A
morfologia celular foi avaliada por microscopia confocal. Resultados: Foi
padronizado um modelo de explante baseado nas diferenças biológicas das
regiões específicas do queloide. A atividade celular em fibroblastos de
queloide variou ao longo do tempo, sendo diferente entre os subgrupos
celulares mas similar dentro de um mesmo subgrupo, mas não foi afetada
pela adição de CGRP, SP ou CGRP+SP. Fibroblastos de queloide das
regiões periférica e central apresentaram maior expressão de p53 do que
fibroblastos da pele adjacente ao queloide, de cicatriz normotrófica ou de
pele normal, adjacente à cicatriz normotrófica. CGRP, SP e CGRP+SP
provocaram uma diminuição na expressão de p53 em fibroblastos de
queloide das regiões periférica e central em relação aos outros subgrupos
estudados. Conclusão: CGRP, SP e CGRP+SP provocaram uma
diminuição na expressão de p53 em fibroblastos de queloide.
Introduction: Given that nociceptive neurogenic stimulation is considered the primary trigger of the healing process in the skin, that CGRP and SP increase fibroblast proliferation in vitro, and given the lack of studies correlating CGRP, SP p53 and apoptosis in keloid, it becomes important to investigate whether skin neuropeptides can influence the expression of p53 and apoptosis in keloid. A possible association between neuropeptides and p53 gene could represent a new path to a better physiopathogenic understanding of pathological fibroproliferative scars and, therefore, the advent of more specific treatments. Objective: To investigate the expression of p53 gene and the apoptotic index in keloid fibroblast cultures exposed to the action of the neuropeptides CGRP and SP. Methods: Keloid fibroblasts were grown in culture medium with or without the addition of CGRP and SP. The apoptotic index and cell cycle were determined by flow cytometry and the labeling of apoptosis key proteins was done by spectrophotometry. Cell activity was evaluated over time by MTT. Cell morphology was analyzed by confocal microscopy. Results: An explant model for keloid fibroblast isolation and culture based on the biological differences of its specific regions was standardized. Cellular activity in keloid fibroblasts varied over time and was different among the fibroblasts subgroups but similar within the same subgroup, however was not affected by the addition of CGRP, SP or SP + CGRP. Keloid fibroblasts derived from peripheral and central regions had higher p53 expression than fibroblasts derived from the skin adjacent to the keloid scar, fibroblasts derived from normotrophic scar or from normal skin adjacent to the normotrophic scar. CGRP, SP and SP + CGRP caused a decrease in p53 expression in keloid fibroblasts of peripheral and central regions compared to all other subgroups studied. Conclusion: CGRP, SP and SP + CGRP caused a decrease in p53 expression in keloid fibroblasts.
Introduction: Given that nociceptive neurogenic stimulation is considered the primary trigger of the healing process in the skin, that CGRP and SP increase fibroblast proliferation in vitro, and given the lack of studies correlating CGRP, SP p53 and apoptosis in keloid, it becomes important to investigate whether skin neuropeptides can influence the expression of p53 and apoptosis in keloid. A possible association between neuropeptides and p53 gene could represent a new path to a better physiopathogenic understanding of pathological fibroproliferative scars and, therefore, the advent of more specific treatments. Objective: To investigate the expression of p53 gene and the apoptotic index in keloid fibroblast cultures exposed to the action of the neuropeptides CGRP and SP. Methods: Keloid fibroblasts were grown in culture medium with or without the addition of CGRP and SP. The apoptotic index and cell cycle were determined by flow cytometry and the labeling of apoptosis key proteins was done by spectrophotometry. Cell activity was evaluated over time by MTT. Cell morphology was analyzed by confocal microscopy. Results: An explant model for keloid fibroblast isolation and culture based on the biological differences of its specific regions was standardized. Cellular activity in keloid fibroblasts varied over time and was different among the fibroblasts subgroups but similar within the same subgroup, however was not affected by the addition of CGRP, SP or SP + CGRP. Keloid fibroblasts derived from peripheral and central regions had higher p53 expression than fibroblasts derived from the skin adjacent to the keloid scar, fibroblasts derived from normotrophic scar or from normal skin adjacent to the normotrophic scar. CGRP, SP and SP + CGRP caused a decrease in p53 expression in keloid fibroblasts of peripheral and central regions compared to all other subgroups studied. Conclusion: CGRP, SP and SP + CGRP caused a decrease in p53 expression in keloid fibroblasts.
Descrição
Citação
TUCCI-VIEGAS, Vanina Monique. CGRP e SP em cultura de fibroblastos de queloide. 2011. 153 f. Tese (Doutorado em Ciências) - Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, 2011.