Flow cytometry, with double staining with Nile red and anti-CD3 antibody, to detect phospholipidosis in peripheral blood lymphocytes of rats treated with amiodarone.
Cell Biol Toxicol
; 25(6): 587-98, 2009 Dec.
Article
em En
| MEDLINE
| ID: mdl-19105033
ABSTRACT
A flow cytometry method, to monitor peripheral lymphocytes phospholipidosis, has been set up using a single staining with Nile red and double staining with Nile red and anti-CD3 monoclonal antibody. Blood has been collected from rats treated with amiodarone (phospholipidogenic antiarrhythmic drug). By flow cytometer, it is possible to detect phospholipids, using Nile red, a probe for intracellular lipids staining, changing its fluorescence on the stained lipid basis. CD3 antigen has been selected to focus on T cells, to evaluate whether these cells are the target of phospholipidosis amiodarone-dependent. In the study A, Sprague-Dawley rats were treated with three different doses (75, 150, and 300 mg kg(-1) day(-1)) of amiodarone or vehicle alone, for 14 days, followed by 14 days of recovery Data obtained show that by flow cytometry, with Nile red alone, it is possible to detect a dose- and time-related response of phospholipidosis-positive lymphocytes; a partial recovery is also assessed. In the study B, Sprague-Dawley rats were treated with a single dose (300 mg kg(-1) day(-1)) of amiodarone, for 14 days Data obtained show that animals treated with amiodarone have a significant increase of phospholipidosis-positive lymphocytes (p = 0.008), in particular of CD3+ cells (p = 0.0056). Transmission electron microscopy analysis confirmed data obtained by flow cytometry. This work shows that flow cytometry with Nile red could be a good tool to monitor ex vivo phospholipidosis in lymphocyte cells of animals treated with amiodarone The phospholipidogenic effect is more evident focusing on CD3+ T lymphocytes, thus suggesting that these cells are probably the target of phospholipidosis.
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1
Base de dados:
MEDLINE
Assunto principal:
Oxazinas
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Fosfolipídeos
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Linfócitos T
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Complexo CD3
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Citometria de Fluxo
Limite:
Animals
Idioma:
En
Ano de publicação:
2009
Tipo de documento:
Article