Regulation of tubular cell MCP-1 production by intracellular ions: a role for sodium and calcium

Exp Nephrol. 2001;9(3):205-13. doi: 10.1159/000052613.

Abstract

Proximal tubule cells (PTC) in chronic renal disease produce chemokines which cause renal interstitial inflammation, and also transport more Na+ than normal. To investigate whether these two events might be related, monocyte chemoattractant protein-1 (MCP-1) production was examined in rat PTC in primary culture. Amiloride reduced, while ouabain increased levels of MCP-1 mRNA and protein. Amiloride reduced MCP-1 in cells stimulated with ouabain, lipopolysaccharide (LPS) or albumin. Intracellular Na+ rose with ouabain, but not LPS or albumin. Effects of amiloride, ouabain, LPS and albumin were abrogated by sodium-free but not chloride-free culture medium, and were not explained by changes in intracellular pH. Intracellular Ca2+ rose with ouabain, LPS or albumin and sodium-free medium. BAPTA-AM reduced intracellular Ca2+ and MCP-1 mRNA levels in unstimulated cells, and cells stimulated with ouabain, LPS or albumin. Thus, amiloride and ouabain may alter tubular cell MCP-1 by changing intracellular Na+, with secondary changes in intracellular Ca2+, whereas stimulation by LPS and albumin may involve Ca2+ directly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid-Base Equilibrium / physiology
  • Amiloride / analogs & derivatives
  • Amiloride / pharmacology*
  • Animals
  • Calcium / metabolism*
  • Cell Survival / physiology
  • Cells, Cultured
  • Chemokine CCL2 / antagonists & inhibitors*
  • Chemokine CCL2 / biosynthesis
  • Chemokine CCL2 / physiology
  • Dose-Response Relationship, Drug
  • Egtazic Acid / analogs & derivatives
  • Egtazic Acid / pharmacology*
  • Enzyme-Linked Immunosorbent Assay
  • Fluorescent Dyes
  • Hydrogen-Ion Concentration
  • Kidney Tubules, Proximal / drug effects*
  • Kidney Tubules, Proximal / metabolism
  • Lipopolysaccharides / pharmacology*
  • Male
  • Ouabain / pharmacology*
  • RNA, Messenger / metabolism*
  • Rats
  • Rats, Wistar
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sodium / metabolism*
  • Sodium-Potassium-Exchanging ATPase / metabolism
  • Spectrometry, Fluorescence

Substances

  • Chemokine CCL2
  • Fluorescent Dyes
  • Lipopolysaccharides
  • RNA, Messenger
  • 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester
  • Egtazic Acid
  • Ouabain
  • Amiloride
  • Sodium
  • Sodium-Potassium-Exchanging ATPase
  • Calcium
  • ethylisopropylamiloride