A new method for detecting Na+ , K+ -ATPase activity by ICP-MS: Quantitative analysis on the inhibitory effect of rhein on Na+ , K+ -ATPase activity by ICP-MS in HCT116 cells

Biomed Chromatogr. 2021 Dec;35(12):e5199. doi: 10.1002/bmc.5199. Epub 2021 Jul 27.

Abstract

Rhein is an active component from Chinese herbal medicine. It can cause diarrhea by inhibiting Na+ , K+ -ATPase activity on intestinal epithelial cells, thus decreasing the re-absorption of Na+ from intestinal tract to blood. However, when this Na+ , K+ -ATPase inhibition was quantitated by a colorimetric method that measures ATPase-catalyzed release of inorganic phosphorus, the data obtained were inconsistent and showed great variation. We developed a novel method using inductively coupled plasma mass spectrometry (ICP-MS) to quantitate the amount of intracellular Rb+ . This method largely mimics the 86 RbCl tracer flux assay, but it uses non-radioactive RbCl as a flux substrate. The results demonstrated that this method has better precision and accuracy than the conventional colorimetric method. More importantly, this method is free from radioactive substances, which is expected to make it safer and more convenient than the radioactive 86 RbCl tracer flux method. In conclusion, the ICP-MS method for Na+ , K+ -ATPase activity determination is novel and accurate. It can also provide a reference for studying the transport of other metal ions across membranes under biological conditions.

Keywords: Na+, K+-ATPase; colorimetric method; inductively coupled plasma mass spectrometry; rhein; rubidium chloride.

MeSH terms

  • Anthraquinones / pharmacology*
  • Chlorides
  • Colorimetry
  • HCT116 Cells
  • Humans
  • Limit of Detection
  • Mass Spectrometry / methods*
  • Reproducibility of Results
  • Rubidium
  • Sodium-Potassium-Exchanging ATPase* / antagonists & inhibitors
  • Sodium-Potassium-Exchanging ATPase* / metabolism

Substances

  • Anthraquinones
  • Chlorides
  • Sodium-Potassium-Exchanging ATPase
  • Rubidium
  • rubidium chloride
  • rhein