Cox-2 and osteopontin in cocultured platelets and mesangial cells: role of glucocorticoids

Kidney Int. 2000 Jun;57(6):2229-38. doi: 10.1046/j.1523-1755.2000.00083.x.


Background: Glomerular inflammation is characterized by a consecutive infiltration of immunoreactive cells. To mimic the early phase of glomerular injury, a coculture system of platelets and rat renal mesangial cells was established. As prototypes, the inflammation-related proteins cyclooxygenase-2 (Cox-2) and the chemotactic protein osteopontin (OPN) were investigated.

Methods: The expression of OPN and Cox-2 mRNA and protein was determined by Northern and Western blot analyses.

Results: Coincubation of platelets and mesangial cells led to a rapid, transient induction of Cox-2 mRNA, which peaked at two hours, whereas OPN and monocyte chemoattractant protein-1 (MCP-1) were induced at later time points. The induction of Cox-2 mRNA was concentration dependent and highly reproducible when platelets of different donors were investigated. Partial Cox-2 induction was observed when supernatants of preactivated platelets were incubated with mesangial cells. The inhibition of the signaling pathways of platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) or interference with Gi-protein signaling partially inhibited platelet-induced Cox-2 expression. Down-regulation of protein kinase C (PKC), which is a common signaling module in many pathways leading to Cox-2 induction, almost completely abrogated platelet-induced Cox-2 expression. The time pattern of Cox-2 and OPN expression suggested that Cox-2 might play a role in OPN induction. The up-regulation of OPN was dependent on de novo protein synthesis and was induced by high levels of exogenous prostaglandin E2 (PGE2; 10 micromol/L). Endogenous PGE2, however, proved not to be essential for OPN mRNA expression, because inhibition of Cox activity did not change OPN mRNA levels. Dexamethasone inhibited Cox-2 mRNA induction but increased OPN mRNA and protein expression.

Conclusion: These data indicate that Cox-2 and OPN are independently up-regulated upon interaction of platelets and mesangial cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Platelets / drug effects
  • Blood Platelets / metabolism*
  • Blood Platelets / physiology
  • Cell Division / physiology
  • Coculture Techniques
  • Cyclooxygenase 2
  • Dexamethasone / pharmacology
  • Gene Expression Regulation / physiology
  • Glomerular Mesangium / cytology
  • Glomerular Mesangium / enzymology*
  • Glomerular Mesangium / physiology
  • Glucocorticoids / pharmacology
  • Glucocorticoids / physiology*
  • Growth Substances / physiology
  • Humans
  • Isoenzymes / blood*
  • Isoenzymes / genetics
  • Membrane Proteins
  • Osteopontin
  • Prostaglandin-Endoperoxide Synthases / blood*
  • Prostaglandin-Endoperoxide Synthases / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Sialoglycoproteins / blood*
  • Sialoglycoproteins / genetics
  • Up-Regulation


  • Glucocorticoids
  • Growth Substances
  • Isoenzymes
  • Membrane Proteins
  • RNA, Messenger
  • SPP1 protein, human
  • Sialoglycoproteins
  • Spp1 protein, rat
  • Osteopontin
  • Dexamethasone
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases