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Creatine supplementation prevents hyperhomocysteinemia, oxidative stress and cancer-induced cachexia progression in Walker-256 tumor-bearing rats

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Abstract

The purpose of this study was to investigate (1) the impact of tumor growth on homocysteine (Hcy) metabolism, liver oxidative stress and cancer cachexia and, (2) the potential benefits of creatine supplementation in Walker-256 tumor-bearing rats. Three experiments were conducted. First, rats were killed on days 5 (D5), 10 (D10) and 14 (D14) after tumor implantation. In experiment 2, rats were randomly assigned to three groups designated as control (C), tumor-bearing (T) and tumor-bearing supplemented with creatine (TCr). A life span experiment was conducted as the third experiment. Creatine was supplied in drinking water for 21 days (8 g/L) in all cases. Tumor implantation consisted of a suspension of Walker-256 cells (8.0 × 107 cells in 0.5 mL of PBS). The progressive increase (P < 0.05) in tumor mass coincided with a progressively lower body weight and higher hepatic oxidative stress; plasma Hcy concentration was 80 % higher (P < 0.05) by 10 days of tumor implantation. Impaired Hcy metabolism was evidenced by decreased hepatic betaine-homocysteine methyltransferase (Bhmt), glycine N-methyltransferase (Gnmt) and cystathionine beta synthase (CBS) gene expression. In contrast, creatine supplementation promoted a 28 % reduction of tumor weight (P < 0.05). Plasma Hcy (C 6.1 ± 0.6, T 10.3 ± 1.5, TCr 6.3 ± 0.9, µmol/L) and hepatic oxidative stress were lower in the TCr group compared to T. Creatine supplementation was unable to decrease Hcy concentration and to increase SAM/SAH ratio in tumor tissue. These data suggest that creatine effects on hepatic impaired Hcy metabolism promoted by tumor cell inoculation are responsible to decrease plasma Hcy in tumor-bearing rats. In conclusion, Walker-256 tumor growth is associated with progressive hyperhomocysteinemia, body weight loss and liver oxidative stress in rats. Creatine supplementation, however, prevented these tumor-associated perturbations.

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Abbreviations

AOPP:

Advanced oxidation protein products

AGAT:

Arginineglycine amidinotransferase

ALT:

Alanine aminotransferase

Bhmt:

Betaine-homocysteine S-methyltransferase

Cbs:

Cystathionine beta synthase

CK:

Creatine kinase

Gnmt:

Glycine N-methyltransferase

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

Hcy:

Homocysteine

HHcy:

Hyperhomocysteinemia

Mat1a:

Methionine adenosyltransferase I alpha

MDA:

Malondialdehyde

Mtrr:

5-methyltetrahydrofolate-homocysteine methyltransferase reductase

Pemt:

Phosphatidylethanolamine N-methyltransferase

SAH:

S-adenosylhomocysteine

SAM:

S-adenosylmethionine

SOD:

Superoxide dismutase

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Acknowledgments

Supported by Brazilian grants from Fapesp (Fundação de Amparo a Pesquisa do estado de São Paulo), CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) and Coordenação de Aperfeiçoamento de Pessoal do Ensino Superior (Capes: 88881.068035/2014-01); and Canadian grants from (Canadian Institutes of Health Research and the Research Development Corporation of Newfoundland and Labrador).

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Authors and Affiliations

  1. Department of Physical Education, Faculty of Physical Education and Sport, State University of Londrina, Rodovia Celso Garcia Cid, Pr 445 km 380, Campus Universitário, Londrina, PR, Brazil

    Rafael Deminice, Paola Sanches Cella & Camila S. Padilha

  2. Department of Pathology, State University of Londrina, Londrina, PR, Brazil

    Fernando H. Borges, Rubens Cecchini & Flávia Alessandra Guarnier

  3. Faculty of Medicine of Ribeirao Preto, University of Sao Paulo, Ribeirão Preto, SP, Brazil

    Lilian Eslaine Costa Mendes da Silva & Alceu Afonso Jordao

  4. Department of Biochemistry, Memorial University of Newfoundland, St. John’s, NL, Canada

    Jason Lorne Robinson & Robert F. Bertolo

  5. School of Physical Education and Sport, University of Sao Paulo, São Paulo, Brazil

    Patrícia L. Campos-Ferraz

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  1. Rafael Deminice
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  2. Paola Sanches Cella
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Correspondence to Rafael Deminice.

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Deminice, R., Cella, P.S., Padilha, C.S. et al. Creatine supplementation prevents hyperhomocysteinemia, oxidative stress and cancer-induced cachexia progression in Walker-256 tumor-bearing rats. Amino Acids 48, 2015–2024 (2016). https://doi.org/10.1007/s00726-016-2172-9

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