Proapoptotic effects of long-chain vitamin E metabolites in HepG2 cells are mediated by oxidative stress

Free Radic Biol Med. 2010 Nov 1;49(8):1315-22. doi: 10.1016/j.freeradbiomed.2010.07.024. Epub 2010 Aug 6.

Abstract

Although the metabolism of vitamin E has been extensively studied in cell culture, animals, and humans, biochemical analyses of intermediate metabolites are scarce. We here describe the synthesis and proapoptotic properties of long-chain metabolites of α- and δ-tocopherol. Several long-chain vitamin E metabolites, namely 13'-hydroxy- and 13'-carboxychromanols, were synthesized from garcinoic acid, a δ-tocotrienol derivative extracted from the African bitter nut Garcinia kola. Both α- and δ-13'-carboxychromanol induced cell death in HepG2 cells at EC(50) of 13.5 and 6.5 μM, respectively. Apoptosis was quantified by annexin V/7-AAD staining and flow cytometry analysis. By immunoblot analyses, we observed activation of both caspase-3 and caspase-9 as well as PARP-1 cleavage. Parameters of mitochondrial dysfunction including reduced mitochondrial membrane potential and increased intracellular and intramitochondrial reactive oxygen species formation were observed after metabolite treatment. Last, long-chain hydroxychromanols were readily metabolized to the corresponding carboxychromanols in HepG2 cells. Taken together, these results indicate that long-chain metabolites may be responsible for antiproliferative properties of vitamin E vitamers.

MeSH terms

  • Apoptosis / drug effects*
  • Benzopyrans / chemical synthesis
  • Benzopyrans / metabolism*
  • Caspase 3 / metabolism
  • Caspase 9 / metabolism
  • Garcinia kola / metabolism
  • Hep G2 Cells
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism
  • Hepatocytes / pathology
  • Humans
  • Membrane Potential, Mitochondrial / drug effects
  • Mitochondria, Liver / drug effects*
  • Mitochondria, Liver / metabolism
  • Mitochondria, Liver / pathology
  • Oxidative Stress
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases / metabolism
  • Reactive Oxygen Species / metabolism
  • Seeds
  • Vitamin E / analogs & derivatives
  • Vitamin E / chemical synthesis
  • Vitamin E / pharmacology*

Substances

  • Benzopyrans
  • Reactive Oxygen Species
  • garcinoic acid
  • Vitamin E
  • PARP1 protein, human
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases
  • Caspase 3
  • Caspase 9