The potent antioxidant activity of the vitamin K cycle in microsomal lipid peroxidation

Biochem Pharmacol. 1997 Oct 15;54(8):871-6. doi: 10.1016/s0006-2952(97)00254-2.


In the vitamin K cycle, vitamin K-hydroquinone, the active cofactor for gamma-glutamylcarboxylase, is continuously regenerated. The successive pathways contain oxidation of the hydroquinone to the epoxide, followed by reduction to the quinone and reduction to the hydroquinone. Vitamin K-hydroquinone is a potent radical scavenging species (Mukai et al., J Biol Chem 267: 22277-22281, 1992). We tested the potential antioxidant activity of the vitamin K cycle in lipid peroxidation reactions (thiobarbituric acid reactive substances, TBARS) in rat liver microsomes. As prooxidant we used Fe2+/ascorbate, NADPH-Fe3+/ATP, and NADPH/CCl4. Vitamin K (< or = 50 microM) on its own did not influence the formation of TBARS. In combination with 1 mM dithiothreitol (DTT), the reductive cofactor for the microsomal enzyme vitamin K epoxide reductase, vitamin K suppressed lipid peroxidation with a concentration that blocked the maximal response by 50% (IC50) of ca. 0.2 microM. Vitamin K1 (phylloquinone) and vitamin K2 (menaquinone-4) were equally active. Warfarin (5 microM) and chloro-vitamin K (50 microM), inhibitors of vitamin K epoxide reductase and gamma-glutamylcarboxylase, respectively, were able to completely abolish the antioxidant effect. Lipid peroxidation was inversely related to the amount of vitamin K hydroquinone in the reaction. Vitamin K epoxide reductase seemed sensitive to lipid peroxidation, with half of the activity being lost within 10 min during oxidation with NADPH/CCl4. The inactivation could be attenuated by antioxidants such as vitamin E, reduced glutathione, and menadione and also by a K vitamin in combination with DTT, but not by superoxide dismutase and catalase. The results show that the vitamin K cycle could act as a potent antioxidant, that the active species in all probability is vitamin K-hydroquinone, and that the primary reaction product is the semiquinone. The results also show that the reaction product is processed in the vitamin K cycle to regenerate vitamin K-hydroquinone.

MeSH terms

  • Animals
  • Antioxidants / metabolism*
  • Cell Compartmentation
  • Dithiothreitol / metabolism
  • Endoplasmic Reticulum, Rough / metabolism
  • Hydroquinones / metabolism
  • Lipid Peroxides / metabolism*
  • Male
  • Microsomes, Liver / metabolism
  • Mixed Function Oxygenases / metabolism*
  • Oxidation-Reduction
  • Oxidative Stress
  • Rats
  • Rats, Wistar
  • Vitamin K / metabolism*
  • Vitamin K Epoxide Reductases


  • Antioxidants
  • Hydroquinones
  • Lipid Peroxides
  • Vitamin K
  • Mixed Function Oxygenases
  • Vitamin K Epoxide Reductases
  • Dithiothreitol