Evans blue dye as a marker of albumin clearance in cultured endothelial monolayer and isolated lung

C. E. Patterson, R. A. Rhoades, J. G.N. Garcia

Research output: Contribution to journalArticle

208 Scopus citations

Abstract

Determination of protein transfer across the endothelial barrier or the entire alveolar capillary membrane is critical for investigation of mechanisms leading to pulmonary edema. The purpose of this study was to evaluate Evans blue dye for determination of protein clearance across cultured bovine pulmonary artery endothelial cell monolayers and as a quantitative marker for albumin leakage to the air spaces in isolated perfused rat lungs. Evans blue dye bound tightly to albumin (EBA) as determined by lack of transfer through dialysis membranes and specific elution with albumin from a molecular exclusion column. EBA was equivalent to 125I-labeled albumin for calculation of albumin clearance rates (C(alb)) across intact and challenged monolayers [C(alb) (+ vehicle) = 0.12 μl/min; C(alb) (+10 nM α-thrombin) = 0.47 μl/min; C(alb) (+5 mg/ml trypsin) = 1.29 μl/min]. Transfer of EBA was linear with time in both the endothelial cell monolayer model and the perfused lung. EBA was a sensitive marker for early edema in the perfused lung (before detectable weight gain) as well as for severe edema in the oxidant-injured lung (marked EBA accumulation in lavage fluid) and was a more specific marker for protein transfer than lavage fluid protein. EBA transfer is a convenient, reproducible, and accurate means to assess alterations in vascular permeability.

Original languageEnglish (US)
Pages (from-to)865-873
Number of pages9
JournalJournal of Applied Physiology
Volume72
Issue number3
DOIs
StatePublished - 1992

Keywords

  • alveolar-capillary membrane permeability
  • endothelial cell monolayer integrity
  • pulmonary edema

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

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