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Astaxanthin prevents in vitro auto-oxidative injury in human lymphocytes

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Abstract

Upon mitogen sensitization, lymphocytes undergo proliferation by oxyradical-based mechanisms. Through continuous resting–restimulation cycles, lymphocytes accumulate auto-induced oxidative lesions which lead to cell dysfunction and limit their viability. Astaxanthin (ASTA) is a nutritional carotenoid that shows notable antioxidant properties. This study aims to evaluate whether the in vitro ASTA treatment can limit oxyradical production and auto-oxidative injury in human lymphocytes. Activated lymphocytes treated with 5 µM ASTA showed immediate lower rates of O •−2 /H2O2 production whilst NO• and intracellular Ca2+ levels were concomitantly enhanced (≤4 h). In long-term treatments (>24 h), the cytotoxicity test for ASTA showed a sigmoidal dose–response curve (LC50 = 11.67 ± 0.42 µM), whereas higher activities of superoxide dismutase and catalase in 5 µM ASTA-treated lymphocytes were associated to significant lower indexes of oxidative injury. On the other hand, lower proliferative scores of ASTA lymphocytes might be a result of diminished intracellular levels of pivotal redox signaling molecules, such as H2O2. Further studies are necessary to establish the ASTA-dose compensation point between minimizing oxidative damages and allowing efficient redox-mediated immune functions, such as proliferation, adhesion, and oxidative burst.

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Abbreviations

ANOVA:

Analysis of variance

ASTA:

Astaxanthin

BHT:

Butylated hydroxytoluene

BSA:

Albumin

[Ca2+]i :

Intracellular calcium

CAT:

Catalase

Con A:

Concanavalin A

DHE:

Dihydroethidium

DMSO:

Dimethyl sulfoxide

DNPH:

2,4-Dinitrophenylhydrazine

DTNB:

5,5′-Dithiobis(2-nitrobenzoic acid)

EDTA:

Ethylenediaminetetraacetic acid

EGTA:

Ethylene glycol tetracetic acid

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

H2O2 :

Hydrogen peroxide

LPS:

Lipopolysaccharide

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NADH:

Nicotinamide adenine dinucleotide

NADPH:

Nicotinamide adenine dinucleotide phosphate

NBT:

Nitroblue tetrazolium

NEM:

N-ethylmalemide

NO:

Nitric oxide

PMA:

Phorbol-12-myristate 13-acetate

PMS:

Phenazine methosulfate

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

TBA:

Thiobarbituric acid

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Acknowledgements

The authors are indebted to the constant assistance of Buttignol MHP, Campoio TR. This research is supported by Fundação do Amparo a Pesquisa do Estado de São Paulo (FAPESP 07/03334-6; 02/09405-9), PIBIC-CNPq (119256/2009-6; 109713/2008-7), and Universidade Cruzeiro do Sul.

Declaration of interest

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

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Authors and Affiliations

  1. Cellular Physiology Laboratory, Postgraduate Program—Health Science, CBS, Cruzeiro do Sul University, Avenida Regente Feijó, 1295, Tatuapé, 03342-000, São Paulo, Brazil

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

  2. Postgraduate Program—Health Science, Cruzeiro do Sul University, Avenida Regente Feijó, 1295, Tatuapé, 03342-000, São Paulo, Brazil

    Marcelo P. Barros & Rosemari Otton

  3. Postgraduate Program—Human Movement Sciences Institute of Physical Activity and Sport Sciences, Cruzeiro do Sul University, Avenida Regente Feijó, 1295, Tatuapé, 03342-000, São Paulo, Brazil

    Marcelo P. Barros & Rosemari Otton

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  1. Anaysa P. Bolin
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  2. Rita C. Macedo
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Correspondence to Rosemari Otton.

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Bolin, A.P., Macedo, R.C., Marin, D.P. et al. Astaxanthin prevents in vitro auto-oxidative injury in human lymphocytes. Cell Biol Toxicol 26, 457–467 (2010). https://doi.org/10.1007/s10565-010-9156-4

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