Skip to main content

Advertisement

SpringerLink
Log in

Astaxanthin addition improves human neutrophils function: in vitro study

  • Original Contribution
  • Published:
European Journal of Nutrition Aims and scope Submit manuscript

Abstract

Purpose

The aim of the present study was to evaluate the in vitro effect of carotenoid astaxanthin (ASTA) on the phagocytic and microbicidal capacities, cytokine release, and reactive oxygen species production in human neutrophils.

Methods

The following parameters were evaluated: cytotoxic effect of ASTA on human neutrophils viability, phagocytic and microbicidal capacities of neutrophils by using Candida albicans assay, intracellular calcium mobilization (Fura 2-AM fluorescent probe), superoxide anion (lucigenin and DHE probes), hydrogen peroxide (H2O2, phenol red), and nitric oxide (NO·) (Griess reagent) production, activities of antioxidant enzymes (total/Mn-SOD, CAT, GPx, and GR), oxidative damages in biomolecules (TBARS assay and carbonyl groups), and cytokine (IL-6 and TNF-alpha) release.

Results

Astaxanthin significantly improves neutrophil phagocytic and microbicidal capacity, and increases the intracellular calcium concentration and NO· production. Both functional parameters were accompanied by a decrease in superoxide anion and hydrogen peroxide and IL-6 and TNF-α production. Oxidative damages in lipids and proteins were significantly decreased after ASTA-treatment.

Conclusions

Taken together our results are supportive to a beneficial effect of astaxanthin-treatment on human neutrophils function as demonstrated by increased phagocytic and fungicide capacity as well as by the reduced superoxide anion and hydrogen peroxide production, however, without affecting neutrophils capacity to kill C. albicans. This process appears to be mediated by calcium released from intracellular storages as well as nitric oxide production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126

    Article  CAS  Google Scholar 

  2. Barrow CJ (2007) Marine nutraceuticals and functional foods. CRC Press, Boca Raton

    Google Scholar 

  3. Baynes JW, Thorpe SR (1999) Role of oxidative stress in diabetic complications: a new perspective on an old paradigm. Diabetes 48:1–9

    Article  CAS  Google Scholar 

  4. Bei L, Hu T, Qian ZM, Shen X (1998) Extracellular Ca2+ regulates the respiratory burst of human neutrophils. Biochim Biophys Acta 1404:475–483

    Article  CAS  Google Scholar 

  5. Biteau B, Labarre J, Toledano MB (2003) ATP-dependent reduction of cysteine-sulphinic acid by S. cerevisiae sulphiredoxin. Nature 425:980–984

    Article  CAS  Google Scholar 

  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  7. Chew BP, Park JS, Wong MW, Wong TS (1999) A comparison of the anticancer activities of dietary beta-carotene, canthaxanthin and astaxanthin in mice in vivo. Anticancer Res 19:1849–1853

    CAS  Google Scholar 

  8. Choi SK, Park YS, Choi DK, Chang HI (2008) Effects of astaxanthin on the production of NO and the expression of COX-2 and iNOS in LPS-stimulated BV2 microglial cells. J Microbiol Biotechnol 18:1990–1996

    CAS  Google Scholar 

  9. Clancy RM, Leszczynska-Piziak J, Abramson SB (1992) Nitric oxide, an endothelial cell relaxation factor, inhibits neutrophil superoxide anion production via a direct action on the NADPH oxidase. J Clin Invest 90:1116–1121

    Article  CAS  Google Scholar 

  10. Davi G, Falco A, Patrono C (2005) Lipid peroxidation in diabetes mellitus. Antioxid Redox Signal 7:256–268

    Article  CAS  Google Scholar 

  11. Ding AH, Nathan CF, Stuehr DJ (1988) Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. Comparison of activating cytokines and evidence for independent production. J Immunol 141:2407–2412

    CAS  Google Scholar 

  12. Ewing JF, Janero DR (1995) Microplate superoxide dismutase assay employing a nonenzymatic superoxide generator. Anal Biochem 232:243–248

    Article  CAS  Google Scholar 

  13. Fassett RG, Coombes JS (2009) Astaxanthin, oxidative stress, inflammation and cardiovascular disease. Future Cardiol 5:333–342

    Article  CAS  Google Scholar 

  14. Fraga CG, Leibovitz BE, Tappel AL (1988) Lipid peroxidation measured as thiobarbituric acid-reactive substances in tissue slices: characterization and comparison with homogenates and microsomes. Free Radic Biol Med 4:155–161

    Article  CAS  Google Scholar 

  15. Galligan C, Yoshimura T (2003) Phenotypic and functional changes of cytokine-activated neutrophils. In: Cassatella MA (ed) The neutrophil. Karger, Basel, pp 24–44

    Chapter  Google Scholar 

  16. Ghosh S, Karin M (2002) Missing pieces in the NF-kappaB puzzle. Cell 109(Suppl):S81–S96

    Article  CAS  Google Scholar 

  17. Grando FC, Felicio CA, Twardowschy A, Paula FM, Batista VG, Fernandes LC, Curi R, Nishiyama A (2009) Modulation of peritoneal macrophage activity by the saturation state of the fatty acid moiety of phosphatidylcholine. Braz J Med Biol Res 42:599–605

    Article  CAS  Google Scholar 

  18. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    CAS  Google Scholar 

  19. Himpens B, Missiaen L, Casteels R (1995) Ca2+ homeostasis in vascular smooth muscle. J Vasc Res 32:207–219

    Article  CAS  Google Scholar 

  20. Hussein G, Sankawa U, Goto H, Matsumoto K, Watanabe H (2006) Astaxanthin, a carotenoid with potential in human health and nutrition. J Nat Prod 69:443–449

    Article  CAS  Google Scholar 

  21. Ikeuchi M, Koyama T, Takahashi J, Yazawa K (2007) Effects of astaxanthin in obese mice fed a high-fat diet. Biosci Biotechnol Biochem 71:893–899

    Article  CAS  Google Scholar 

  22. Jyonouchi H, Hill RJ, Tomita Y, Good RA (1991) Studies of immunomodulating actions of carotenoids. I. Effects of beta-carotene and astaxanthin on murine lymphocyte functions and cell surface marker expression in in vitro culture system. Nutr Cancer 16:93–105

    Article  CAS  Google Scholar 

  23. Jyonouchi H, Sun S, Gross M (1995) Effect of carotenoids on in vitro immunoglobulin production by human peripheral blood mononuclear cells: astaxanthin, a carotenoid without vitamin A activity, enhances in vitro immunoglobulin production in response to a T-dependent stimulant and antigen. Nutr Cancer 23:171–183

    Article  CAS  Google Scholar 

  24. Jyonouchi H, Sun S, Tomita Y, Gross MD (1995) Astaxanthin, a carotenoid without vitamin A activity, augments antibody responses in cultures including T-helper cell clones and suboptimal doses of antigen. J Nutr 125:2483–2492

    CAS  Google Scholar 

  25. Jyonouchi H, Zhang L, Gross M, Tomita Y (1994) Immunomodulating actions of carotenoids: enhancement of in vivo and in vitro antibody production to T-dependent antigens. Nutr Cancer 21:47–58

    Article  CAS  Google Scholar 

  26. Kobayashi M (2000) In vivo antioxidant role of astaxanthin under oxidative stress in the green alga Haematococcus pluvialis. Appl Microbiol Biotechnol 54:550–555

    Article  CAS  Google Scholar 

  27. Levy J (1999) Abnormal cell calcium homeostasis in type 2 diabetes mellitus: a new look on old disease. Endocrine 10:1–6

    Article  CAS  Google Scholar 

  28. Liu X, Shibata T, Hisaka S, Osawa T (2009) Astaxanthin inhibits reactive oxygen species-mediated cellular toxicity in dopaminergic SH-SY5Y cells via mitochondria-targeted protective mechanism. Brain Res 1254:18–27

    Article  CAS  Google Scholar 

  29. Lordan S, O’Neill C, O’Brien NM (2008) Effects of apigenin, lycopene and astaxanthin on 7 beta-hydroxycholesterol-induced apoptosis and Akt phosphorylation in U937 cells. Br J Nutr 100:287–296

    Article  CAS  Google Scholar 

  30. Mannervik B (1985) Glutathione peroxidase. Methods Enzymol 113:490–495

    Article  CAS  Google Scholar 

  31. Medina-Silva R, Barros MP, Galhardo RS, Netto LE, Colepicolo P, Menck CF (2006) Heat stress promotes mitochondrial instability and oxidative responses in yeast deficient in thiazole biosynthesis. Res Microbiol 157:275–281

    Article  CAS  Google Scholar 

  32. Mitra S, Abraham E (2006) Participation of superoxide in neutrophil activation and cytokine production. Biochim Biophys Acta 1762:732–741

    CAS  Google Scholar 

  33. Murphy ME, Kehrer JP (1989) Oxidation state of tissue thiol groups and content of protein carbonyl groups in chickens with inherited muscular dystrophy. Biochem J 260:359–364

    CAS  Google Scholar 

  34. Naito Y, Uchiyama K, Aoi W, Hasegawa G, Nakamura N, Yoshida N, Maoka T, Takahashi J, Yoshikawa T (2004) Prevention of diabetic nephropathy by treatment with astaxanthin in diabetic db/db mice. Biofactors 20:49–59

    Article  CAS  Google Scholar 

  35. Newsholme P, Haber EP, Hirabara SM, Rebelato EL, Procopio J, Morgan D, Oliveira-Emilio HC, Carpinelli AR, Curi R (2007) Diabetes associated cell stress and dysfunction: role of mitochondrial and non-mitochondrial ROS production and activity. J Physiol 583:9–24

    Article  CAS  Google Scholar 

  36. Oliveira-Souza M, De Mello-Aires M (2000) Interaction of angiotensin II and atrial natriuretic peptide on pH(i) regulation in MDCK cells. Am J Physiol Renal Physiol 279:F944–F953

    CAS  Google Scholar 

  37. Pashkow FJ, Watumull DG, Campbell CL (2008) Astaxanthin: a novel potential treatment for oxidative stress and inflammation in cardiovascular disease. Am J Cardiol 101:58D–68D

    Article  CAS  Google Scholar 

  38. Peng HB, Rajavashisth TB, Libby P, Liao JK (1995) Nitric oxide inhibits macrophage-colony stimulating factor gene transcription in vascular endothelial cells. J Biol Chem 270:17050–17055

    Article  CAS  Google Scholar 

  39. Pick E, Mizel D (1981) Rapid microassays for the measurement of superoxide and hydrogen peroxide production by macrophages in culture using an automatic enzyme immunoassay reader. J Immunol Methods 46:211–226

    Article  CAS  Google Scholar 

  40. Putney JW Jr, Bird GS (1993) The signal for capacitative calcium entry. Cell 75:199–201

    Article  CAS  Google Scholar 

  41. Rubbo H, Radi R, Trujillo M, Telleri R, Kalyanaraman B, Barnes S, Kirk M, Freeman BA (1994) Nitric oxide regulation of superoxide and peroxynitrite-dependent lipid peroxidation. Formation of novel nitrogen-containing oxidized lipid derivatives. J Biol Chem 269:26066–26075

    CAS  Google Scholar 

  42. Sampaio SC, Sousa-e-Silva MC, Borelli P, Curi R, Cury Y (2001) Crotalus durissus terrificus snake venom regulates macrophage metabolism and function. J Leukoc Biol 70:551–558

    CAS  Google Scholar 

  43. Spiller GA, Dewell A (2003) Safety of an astaxanthin-rich Haematococcus pluvialis algal extract: a randomized clinical trial. J Med Food 6:51–56

    Article  CAS  Google Scholar 

  44. Stegenga ME, Van Der Crabben SN, Dessing MC, Pater JM, van den Pangaart PS, de Vos AF, Tanck MW, Roos D, Sauerwein HP, van der Poll T (2008) Effect of acute hyperglycaemia and/or hyperinsulinaemia on proinflammatory gene expression, cytokine production and neutrophil function in humans. Diabet Med 25:157–164

    Article  CAS  Google Scholar 

  45. Stone JR, Yang S (2006) Hydrogen peroxide: a signaling messenger. Antioxid Redox Signal 8:243–270

    Article  CAS  Google Scholar 

  46. Thomas AP, Bird GS, Hajnoczky G, Robb-Gaspers LD, Putney JW Jr (1996) Spatial and temporal aspects of cellular calcium signaling. FASEB J 10:1505–1517

    CAS  Google Scholar 

  47. Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J (2007) Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 39:44–84

    Article  CAS  Google Scholar 

  48. Yanagisawa S, Koarai A, Sugiura H, Ichikawa T, Kanda M, Tanaka R, Akamatsu K, Hirano T, Matsunaga K, Minakata Y, Ichinose M (2009) Oxidative stress augments toll-like receptor 8 mediated neutrophilic responses in healthy subjects. Respir Res 10:50

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors are indebted to the constant assistance of Sampaio, SC, Buttignol MHP, Campoio TR and the human volunteers. This research is supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP 07/03334-6 and 08/50066-0), and Universidade Cruzeiro do Sul.

Conflict of interest statement

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Author information

Authors and Affiliations

  1. Postgraduate Program, Health Science, CBS, Cruzeiro do Sul University, Avenida Regente Feijó, 1295. Tatuapé, São Paulo, SP, CEP 03342-000, Brazil

    Rita C. Macedo, Anaysa P. Bolin, Douglas P. Marin & Rosemari Otton

  2. Postgraduate Program, Human Movement Sciences Institute of Physical Activity and Sport Sciences, ICAFE, Cruzeiro do Sul University, São Paulo, SP, 01506-000, Brazil

    Rosemari Otton

Authors
  1. Rita C. Macedo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anaysa P. Bolin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Douglas P. Marin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rosemari Otton
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rosemari Otton.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Macedo, R.C., Bolin, A.P., Marin, D.P. et al. Astaxanthin addition improves human neutrophils function: in vitro study. Eur J Nutr 49, 447–457 (2010). https://doi.org/10.1007/s00394-010-0103-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00394-010-0103-1

Keywords

Search

Navigation