Aminaftone, a derivative of 4-aminobenzoic acid, downregulates endothelin-1 production in ECV304 Cells: an in vitro Study

Drugs R D. 2008;9(4):251-7. doi: 10.2165/00126839-200809040-00005.

Abstract

Background and objective: Endothelin-1 (ET-1) plays a central role in the pathogenesis of several vascular diseases. Aminaftone is a drug used for the treatment of capillary disorders but which has a mechanism of action that is not fully understood. We investigated whether aminaftone may exert its effect by interfering with the production of ET-1.

Methods: Human ECV304 endothelial cells were incubated with interleukin-1beta (IL-1beta) 100 IU/mL with or without the addition of increasing concentrations of aminaftone (2, 4 or 6 microg/mL). ET-1 concentrations in surnatants were quantified by enzyme immunoassay kit at 3, 6 and 12 hours. Pre-pro-endothelin-1 (PPET-1) gene expressions were also analysed by real-time polymerase chain reaction (RT-PCR) at the same time points. Endothelin-converting enzyme (ECE) activity was also determined.

Results: Incubation with IL-1beta increased concentrations of ET-1 and PPET-1 relative gene expression. Incubation with aminaftone significantly reduced production of ET-1 in a concentration-dependent manner. A strong direct correlation was found between ET-1 concentrations and PPET-1 relative gene expression, but aminaftone did not influence ECE activity.

Conclusion: Aminaftone inhibits ET-1 production in cell cultures by interfering with transcription of the PPET-1 gene. These findings may account for the clinical efficacy of aminaftone in the treatment of capillary disorders and may encourage conduct of further clinical trials.

MeSH terms

  • 4-Aminobenzoic Acid / pharmacology
  • Aspartic Acid Endopeptidases / biosynthesis
  • Aspartic Acid Endopeptidases / genetics
  • Cell Line
  • Down-Regulation / drug effects
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Endothelin-1 / biosynthesis*
  • Endothelin-Converting Enzymes
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Hemostatics / pharmacology*
  • Humans
  • Interleukin-1beta / blood
  • Metalloendopeptidases / biosynthesis
  • Metalloendopeptidases / genetics
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • para-Aminobenzoates*

Substances

  • Endothelin-1
  • Hemostatics
  • Interleukin-1beta
  • RNA, Messenger
  • para-Aminobenzoates
  • aminaftone
  • Aspartic Acid Endopeptidases
  • Metalloendopeptidases
  • Endothelin-Converting Enzymes
  • 4-Aminobenzoic Acid