The CB1/VR1 agonist arvanil induces apoptosis through an FADD/caspase-8-dependent pathway

Br J Pharmacol. 2003 Nov;140(6):1035-44. doi: 10.1038/sj.bjp.0705532. Epub 2003 Oct 6.

Abstract

1. Arvanil (N-arachidonoylvanillamine), a nonpungent capsaicin-anandamide hybrid molecule, has been shown to exert biological activities through VR1/CB1-dependent and -independent pathways. We have found that arvanil induces dose-dependent apoptosis in the lymphoid Jurkat T-cell line, but not in peripheral blood T lymphocytes. Apoptosis was assessed by DNA fragmentation through cell cycle and TUNEL analyses. 2. Arvanil-induced apoptosis was initiated independently of any specific phase of the cell cycle, and it was inhibited by specific caspase-8 and -3 inhibitors and by the activation of protein kinase C. In addition, kinetic analysis by Western blots and fluorimetry showed that arvanil rapidly activates caspase-8, -7 and -3, and induces PARP cleavage. 3. The arvanil-mediated apoptotic response was greatly inhibited in the Jurkat-FADDDN cell line, which constitutively expresses a negative dominant form of the adapter molecule Fas-associated death domain (FADD). This cell line does not undergo apoptosis in response to Fas (CD95) stimulation. 4. Using a cytofluorimetric approach, we have found that arvanil induced the production of reactive oxygen species (ROS) in both Jurkat-FADD+ and Jurkat-FADDDN cell lines. However, ROS accumulation only plays a residual role in arvanil-induced apoptosis. 5. These results demonstrate that arvanil-induced apoptosis is essentially mediated through a mechanism that is typical of type II cells, and implicates the death-inducing signalling complex and the activation of caspase-8. This arvanil-apoptotic activity is TRPV1 and CB-independent, and can be of importance for the development of potential anti-inflammatory and antitumoral drugs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Apoptosis / drug effects*
  • Apoptosis / genetics
  • Capsaicin / analogs & derivatives*
  • Capsaicin / pharmacology*
  • Carrier Proteins / metabolism*
  • Caspase 8
  • Caspase Inhibitors
  • Caspases / metabolism*
  • Cells, Cultured
  • Cysteine Proteinase Inhibitors / pharmacology
  • DNA Fragmentation / drug effects
  • Dose-Response Relationship, Drug
  • Fas Ligand Protein
  • Fas-Associated Death Domain Protein
  • Humans
  • In Situ Nick-End Labeling
  • Jurkat Cells
  • Membrane Glycoproteins / genetics
  • Models, Biological
  • Reactive Oxygen Species / metabolism
  • Receptor, Cannabinoid, CB1 / agonists
  • Receptor, Cannabinoid, CB1 / physiology
  • Receptors, Drug / agonists
  • Receptors, Drug / physiology
  • Signal Transduction*
  • T-Lymphocytes / drug effects
  • Time Factors
  • Transcription, Genetic / drug effects

Substances

  • Adaptor Proteins, Signal Transducing
  • Carrier Proteins
  • Caspase Inhibitors
  • Cysteine Proteinase Inhibitors
  • FADD protein, human
  • FASLG protein, human
  • Fas Ligand Protein
  • Fas-Associated Death Domain Protein
  • Membrane Glycoproteins
  • Reactive Oxygen Species
  • Receptor, Cannabinoid, CB1
  • Receptors, Drug
  • arvanil
  • CASP8 protein, human
  • Caspase 8
  • Caspases
  • Capsaicin