Comparison of formation of D2/E2-isoprostanes and F2-isoprostanes in vitro and in vivo--effects of oxygen tension and glutathione

Arch Biochem Biophys. 1998 May 1;353(1):160-71. doi: 10.1006/abbi.1998.0645.


The isoprostanes (IsoPs) are bioactive prostaglandin-like compounds derived from the free-radical-catalyzed peroxidation of arachidonic acid in vitro and in vivo. IsoPs possessing either an F-type prostane ring (F2-IsoPs) or D/E-type prostane rings (D2/E2-IsoPs) are formed depending on whether IsoP endoperoxide intermediates undergo reduction or isomerization, respectively. Little, however, is known regarding factors influencing the formation of various classes of IsoPs, particularly D2/E2-IsoPs. Thus, studies were undertaken to examine the formation of D2/E2-IsoPs in relation to F2-Isops both in vitro and in vivo. In peroxidizing rat liver microsomes, the formation of D2/E2-IsoPs increased in a time- and oxygen-dependent manner and correlated with F2-IsoP generation and loss of precursor arachidonic acid, although the absolute amount of D2/E2-IsoPs formed exceeded by over 5-fold the levels of F2-IsoPs formed. Surprisingly, however, in liver tissue from rats exposed to an oxidant stress, levels of F2-IsoPs were up to 10-fold greater than those of D2/E2-IsoPs, suggesting that an endogenous process causes IsoP endoperoxide reduction in vivo. Addition of glutathione (GSH) to peroxidizing microsomes at concentrations from 0.01 to 5 mM increased the formation of F2-IsoPs at the expense of D2/E2-IsoPs. Boiling of microsomes did not alter the effect of GSH. Formation of D2/E2-IsoPs in liver tissue in vivo was greatly enhanced compared to F2-IsoPs in rats depleted of GSH. Thus, GSH modulates the formation of different classes of IsoPs in vitro and in vivo. Other thiols, including beta-mercaptoethanol, dithiothreitol, and cysteine, were able to substitute for GSH. These studies indicate that GSH promotes F2-IsoP formation and diminishes D2/E2-IsoP levels in vitro and in vivo by causing reduction of IsoP endoperoxides.

Publication types

  • Comparative Study

MeSH terms

  • Adenosine Diphosphate / pharmacology
  • Animals
  • Ascorbic Acid / pharmacology
  • Carbon Tetrachloride / pharmacology
  • Dinoprost / analogs & derivatives*
  • Dinoprost / metabolism
  • Dinoprostone / analogs & derivatives*
  • Dinoprostone / metabolism
  • Glutathione / metabolism
  • Glutathione Peroxidase / genetics
  • Glutathione Peroxidase / metabolism
  • Humans
  • Iron / pharmacology
  • Kidney / metabolism
  • Lipid Peroxidation*
  • Liver / metabolism*
  • Male
  • Malondialdehyde / analysis
  • Mice
  • Mice, Transgenic
  • Microsomes / metabolism
  • Microsomes, Liver / metabolism*
  • Oxygen / pharmacology
  • Partial Pressure
  • Phospholipids / isolation & purification
  • Phospholipids / metabolism*
  • Prostaglandin D2 / analogs & derivatives*
  • Prostaglandin D2 / metabolism
  • Rats
  • Rats, Sprague-Dawley


  • Phospholipids
  • Malondialdehyde
  • Adenosine Diphosphate
  • Dinoprost
  • Carbon Tetrachloride
  • Iron
  • Glutathione Peroxidase
  • Glutathione
  • Dinoprostone
  • Ascorbic Acid
  • Prostaglandin D2
  • Oxygen