Different Storing and Processing Conditions of Human Lymphocytes do not Alter P-Glycoprotein Rhodamine 123 Efflux.

  • Gemma Chiva-Blanch Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain. 2 Departament de Ciències Fisiològiques II, Facultat de Medicina, IDIBELL, University of Barcelona, Spain.
  • Pepita Giménez-Bonafe 2 Departament de Ciències Fisiològiques II, Facultat de Medicina, IDIBELL, University of Barcelona, Spain.
  • Inés Llaudó 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
  • Joan Torras 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
  • Josep M Cruzado 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
  • Jordi Pétriz 3 Institut de Recerca de l'Hospital Universitari Vall d'Hebron, Barcelona, Spain.
  • Esther Castaño 4 Serveis Cièntifico-tècnics de la Universitat de Barcelona, Unitat de Biologia-Bellvitge.
  • Marcella Franquesa 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
  • Avelina Tortosa 5 School of Nursing, Department of Basic Nursing, IDIBELL–Barcelona University, L'Hospitalet de Llobregat, Spain.
  • Immaculada Herrero-Fresneda 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
  • Inés Rama 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
  • Oriol Bestard 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
  • Josep M Grinyó 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
  • Núria LLoberas 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.

Abstract

P-glycoprotein (Pgp), a protein codified by Multi Drug Resistance (MDR1) gene, has a detoxifying function and might influence the toxicity and pharmacokinetics and pharmacodynamics of drugs. Sampling strategies to improve Pgp studies could be useful to optimize the sensitivity and the reproducibility of efflux assays. This study aimed to compare Pgp expression and efflux activity by measuring Rhodamine123 (Rh123) retention in lymphocytes stored under different conditions, in order to evaluate the potential utility of any of the storing conditions in Pgp functionality. Our results show no change in protein expression of Pgp by confocal studies and Western blotting, nor changes at the mRNA level (qRT-PCR). No differences in Rh123 efflux by Pgp activity assays were found between fresh and frozen lymphocytes after 24 hours of blood extraction, using either of the two Pgp specific inhibitors (VP and PSC833). Different working conditions in the 24 hours post blood extraction do not affect Rh123 efflux. These results allow standardization of Pgp activity measurement in different individuals with different timing of blood sampling and in different geographic areas.

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Author Biographies

Gemma Chiva-Blanch, Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain. 2 Departament de Ciències Fisiològiques II, Facultat de Medicina, IDIBELL, University of Barcelona, Spain.
Nephrology Department Nephrology. BSc
Pepita Giménez-Bonafe, 2 Departament de Ciències Fisiològiques II, Facultat de Medicina, IDIBELL, University of Barcelona, Spain.
Departament de Ciències Fisiològiques II. PhD
Inés Llaudó, 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
Nephrology Department Nephrology. BSc
Joan Torras, 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
Nephrology Department Nephrology. MD, PhD
Josep M Cruzado, 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
Nephrology Department Nephrology. MD, PhD
Marcella Franquesa, 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
Nephrology Department Nephrology. PhD
Immaculada Herrero-Fresneda, 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
Nephrology Department Nephrology. PhD
Oriol Bestard, 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
Nephrology Department Nephrology. MD PhD
Josep M Grinyó, 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
Nephrology Department Nephrology. Prof. MD PhD
Núria LLoberas, 1 Nephrology Department, IDIBELL-Hospital Universitari Bellvitge, L'Hospitalet de Llobregat, Spain.
Nephrology Department Nephrology. PhD
Published
2009-11-30
How to Cite
Chiva-Blanch, G., Giménez-Bonafe, P., Llaudó, I., Torras, J., Cruzado, J., Pétriz, J., Castaño, E., Franquesa, M., Tortosa, A., Herrero-Fresneda, I., Rama, I., Bestard, O., Grinyó, J., & LLoberas, N. (2009). Different Storing and Processing Conditions of Human Lymphocytes do not Alter P-Glycoprotein Rhodamine 123 Efflux. Journal of Pharmacy & Pharmaceutical Sciences, 12(3), 357-366. https://doi.org/10.18433/J34G69
Section
Pharmaceutical Sciences; Review Articles