Endothelin-induced changes of secondary messengers in cultured corneal endothelial cells

J Ocul Pharmacol Ther. 2001 Aug;17(4):351-61. doi: 10.1089/108076801753162762.


The effect of endothelins on corneal endothelial cells is not well understood. We have investigated the effects of endothelin-1 (ET-1), endothelin-2 (ET-2) and endothelin-3 (ET-3) on bovine corneal endothelial cellular proliferation and the secondary messenger changes in cells in the presence of ET-1. It was found that the 3H-thymidine uptake was enhanced by ET-1 significantly, whereas ET-2 and ET-3 had no effect. ET-1 remarkably affects the increase of corneal endothelial cells on 3H-thymidine, 3H-leucine, and 3H-uridine uptakes in a dose-dependent manner. The 50% effective concentrations (EC50) for ET-1, as measured by 3H-thymidine uptake, 3H-uridine uptake, and 3H-leucine uptake were 10(-8.78) M, 10(-8.53) M and 10(-8.04) M, respectively. It was found that endothelin-1 increased intracellular calcium concentration by using the method of preloading with Fura-2-AM and assaying with spectrophotometry. The cellular IP1, IP2, and IP3 were also stimulated in the presence of ET-1. Moreover, ET-1 enhanced the basal cellular cAMP and cGMP concentrations in corneal endothelial cells in a dose-dependent manner. Immunofluorescent staining revealed that ET-1 increased the fibronectin protein concentration and changed protein distribution in corneal endothelial cells. These findings indicate that endothelin-1 increases in cell proliferation and biological changes may be involved in changing intracellular calcium mobility, increasing intracellular phosphoinositides, enhancing intracellular cGMP and cAMP accumulation, and fibronectin protein synthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism
  • Cattle
  • Cell Division / drug effects
  • Cells, Cultured
  • Cyclic AMP / metabolism
  • Cyclic GMP / metabolism
  • DNA Replication
  • Dose-Response Relationship, Drug
  • Endothelin-1 / pharmacology*
  • Endothelium, Corneal / cytology
  • Endothelium, Corneal / drug effects*
  • Endothelium, Corneal / metabolism
  • Fibronectins / metabolism
  • Fura-2 / analogs & derivatives*
  • Fura-2 / metabolism
  • Inositol Phosphates / metabolism
  • Leucine / metabolism
  • Second Messenger Systems* / physiology
  • Thymidine / metabolism
  • Uridine / metabolism


  • Endothelin-1
  • Fibronectins
  • Inositol Phosphates
  • fura-2-am
  • Cyclic AMP
  • Leucine
  • Cyclic GMP
  • Calcium
  • Fura-2
  • Thymidine
  • Uridine