Astaxanthin and omega-3 fatty acids individually and in combination protect against oxidative stress via the Nrf2-ARE pathway

Food Chem Toxicol. 2013 Dec;62:869-75. doi: 10.1016/j.fct.2013.10.023. Epub 2013 Oct 21.


Oxidative stress is a major driver of many diseases, including cancer. The induction of Nrf2-ARE-mediated antioxidant enzymes provides a cellular defense against oxidative stress. Astaxanthin (AST), a red dietary carotenoid, possesses potent antioxidant activity, and inhibits oxidative damages. Polyunsaturated fatty acids (PUFAs), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), are important nutritional essentials and potent antioxidants found in fish oil. In the present study, we investigated whether AST in combination with low concentrations of DHA or EPA has a synergistic antioxidant effect in a HepG2-C8-ARE-luciferase cell line system. Using free radical scavenging DPPH assay, AST was more potent DPPH radical scavenger than DHA and EPA. MTS assay revealed that AST was non-toxic up to 100μM compared with more toxic DHA and EPA. The three compounds alone and in combination elevated cellular GSH levels, increased the total antioxidant activity, induced mRNA expression of Nrf2 and Nrf2 downstream target genes NQO1, HO-1, and GSTM2. Lower concentrations of AST show synergistic effects when combined with DHA or EPA. In summary, our study shows synergistic antioxidant effects of AST and PUFAs at low concentrations. The Nrf2/ARE pathway plays an important role in the antioxidative effects induced by AST, DHA, and EPA.

Keywords: 2,2-diphenyl-1-picrylhydrazyl; ARE; AST; Antioxidant response element; Astaxanthin; CI; DHA; DMSO; DPPH; Docosahexaenoic acid; EPA; Eicosapentaenoic acid; FBS; GSH; GSTM2; HO-1; NAD(P)H: quinone oxydoreductase 1; NQO1; Nrf2; Nuclear factor (erythroid-derived 2)-like 2; OD; Oxidative stress; PUFAs; RSA; SFN; antioxidant response element; astaxanthin; combination index; dimethyl sulfoxide; docosahexaenoic acid; eicosapentaenoic acid; fetal bovine serum; glutathione; glutathione S-transferase M2; heme oxygenase-1; nuclear factor (erythroid-derived 2)-like 2; optical density; polyunsaturated fatty acids; qRT–PCR; quantitative reverse-transcriptase polymerase chain reaction; radical scavenging activity; sulforaphane.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidant Response Elements / drug effects*
  • Antioxidants / pharmacology
  • Dose-Response Relationship, Drug
  • Drug Synergism
  • Fatty Acids, Omega-3 / pharmacology*
  • Glutathione / metabolism
  • Glutathione Transferase / genetics
  • Heme Oxygenase-1 / genetics
  • Hep G2 Cells / drug effects
  • Humans
  • NAD(P)H Dehydrogenase (Quinone) / genetics
  • NF-E2-Related Factor 2 / metabolism*
  • Oxidative Stress / drug effects*
  • Signal Transduction
  • Xanthophylls / pharmacology


  • Antioxidants
  • Fatty Acids, Omega-3
  • NF-E2-Related Factor 2
  • NFE2L2 protein, human
  • Xanthophylls
  • astaxanthine
  • HMOX1 protein, human
  • Heme Oxygenase-1
  • NAD(P)H Dehydrogenase (Quinone)
  • NQO1 protein, human
  • Glutathione Transferase
  • glutathione S-transferase Mu 2
  • Glutathione