Na/K-ATPase assay in the intact mice lung subjected to perfusion

BMC Res Notes. 2014 Nov 15:7:798. doi: 10.1186/1756-0500-7-798.

Abstract

Background: Among the characteristics of acute respiratory distress syndrome (ARDS) is edema formation and its resolution depends on pneumocyte Na/K-ATPase activity. Increased concentration of oleic acid (OA) in plasma induces lung injury by targeting Na/K-ATPase and, thus, interfering in sodium transport.

Findings: Presently, we adapted a radioactivity-free assay to detect Na/K-ATPase activity in perfused lung mice, comparing the inhibitory effect of ouabain and OA. We managed to perfuse only the lung, avoiding the systemic loss of rubidium. Rb+ incorporation into lung was measured by inductively coupled plasma optical emission spectrometry (ICP OES) technique, after lung tissue digestion. Na/K-ATPase activity was the difference between Rb+ incorporation with or without ouabain. Lung Na/K-ATPase was completely inhibited by perfusion with ouabain. However, OA caused a partial inhibition.

Conclusions: In the present work the amount of incorporated Rb+ was greater than seen in our previous report, showing that the present technique is trustworthy. This new proposed assay may allow researchers to study the importance of Na/K-ATPase activity in lung pathophysiology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Enzyme Assays / methods*
  • Lung / drug effects
  • Lung / enzymology*
  • Male
  • Mice
  • Okadaic Acid / pharmacology
  • Ouabain / pharmacology
  • Perfusion*
  • Rubidium / metabolism
  • Sodium-Potassium-Exchanging ATPase / metabolism*

Substances

  • Okadaic Acid
  • Ouabain
  • Sodium-Potassium-Exchanging ATPase
  • Rubidium