In vitro study of P-glycoprotein induction as an antidotal pathway to prevent cytotoxicity in Caco-2 cells

• Tipo: Artigo em Revista Científica Internacional
• Data: 2011

Autores

Renata Silva (Autor) Outra; Helena Carmo (Autor) FFUP; Ricardo Dinis Oliveira (Autor) FMUP; Anabela Cordeiro da Silva (Autor) FFUP; Sofia Costa Lima (Autor) Outra; Felix Carvalho (Autor) FFUP; Maria de Lourdes Bastos (Autor) FFUP; Fernando Remiao (Autor) FFUP

Revista

Título: Archives of Toxicology

Vol. 85

Páginas: 315-326

ISSN: 0340-5761

Editora: Springer Nature

Indexação

ISI Web of Knowledge - 44 Citações

Scopus - 50 Citações

Classificação Científica

FOS: Ciências médicas e da saúde > Medicina básica

Outras Informações

ID Authenticus: P-002-STX

DOI: 10.1007/s00204-010-0587-8

Abstract (EN): The Caco-2 cell line is a reliable in vitro model for predicting drug intestinal absorption and P-glycoprotein (P-gp)-mediated excretion in humans. Recent in vivo studies suggested the induction of P-gp as a cellular protection tool against paraquat poisoning, through the increase in its pulmonary and intestinal excretion. Thus, the aim of the present work was to evaluate P-gp expression and activity in Caco-2 cells exposed to doxorubicin (a known P-gp inducer) and to correlate these changes with paraquat toxic effects. Cytotoxicity of doxorubicin (0-100 mu M) and paraquat (0-1,000 mu M) was evaluated for a maximum period of 96 h. In doxorubicin-exposed cells, P-gp expression and transport activity were evaluated by flow cytometry, using a fluorescein isothiocyanate-conjugated antibody and the P-gp fluorescent subtract rhodamine 123, respectively. A significant increase in P-gp expression was observed as soon as 6 h after exposure to 5 mu M doxorubicin. P-gp activity also increased after 6 h, but only at higher doxorubicin concentrations (over 50 mu M). Paraquat (0-5,000 mu M) cytotoxicity was then evaluated with or without previous exposure of the cells to doxorubicin (5-100 mu M, a concentration range causing both an increase in P-gp expression and activity). Under P-gp induction, a significant reduction in paraquat cytotoxicity was observed. Furthermore, when these cells were incubated with a specific P-gp inhibitor (UIC2 antibody) the doxorubicin protective effects were blocked, confirming the involvement of P-gp in the reduction in paraquat cytotoxicity. In conclusion, the human Caco-2 cell line model can be used for the study of P-gp induction as an antidotal pathway against substrates of this transporter system.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 12