Co-administration of ascorbic acid and α-tocopherol modifies ascorbic acid and attenuates p38, Akt, and TNF-α expression in spinal cord of rats with neuropathic pain

Abstract

Purpose

p38 and Akt plays an important role in neuropathic pain. p38 appears to regulate the SVCT-2 expression, a specific transporter for ascorbic acid (an important antioxidant and neuromodulator). Vitamins C and E induce antinociception and appear to affect p38, Akt, and SVCT-2, but it is unknown their effect on these molecules in spinal cord of rats with chronic constriction injury (CCI) of the sciatic nerve, a model of neuropathic pain. So, we investigated the effect of vitamins C + E on the expression of p38, Akt, and transporter SVCT-2 and ascorbic acid content in the lumbosacral spinal cord of rats with CCI. TNF-α mRNA level was also determined.

Methods

Placement of four loose chromic thread ligatures around the sciatic nerve produced CCI. Ascorbic acid (vitamin C 30 mg/kg/day) + α-tocopherol (vitamin E 15 mg/kg/day) or vehicle (saline containing 1% Tween 80) were administrated (intraperitoneally) daily after CCI for 3 or 10 days. At the end of these periods, lumbosacral spinal cord was dissected out and used for assays.

Results

The vitamins prevented the increase in phosphorylated p38 and phosphorylated Akt and the decrease in SVCT-2 expressions, which were found in vehicle-treated CCI rats at days 3 and 10. Akt expression was reduced only at day 10. Vitamins also prevented the increase in ascorbic acid content and TNF-α level, which was found in vehicle-treated CCI rats.

Conclusions

The changes in p-p38, p-Akt, ascorbic acid, SVCT-2, and TNF-α may be contributing to antinociception induced by vitamins C + E in rats with CCI.

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Data availability

All data generated or analyzed in this study are included in this published article.

Abbreviations

AA:

Ascorbic acid

Akt:

Protein kinase B

ANOVA:

Analysis of variance

CCI:

Chronic constriction injury

CEUA:

Ethics Committee of the Universidade Federal do Rio Grande do Sul

DNPH:

2,4-Dinitrophenylhydrazine

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

I.P. :

Intraperitoneal

KCl:

Potassium chloride

LPS:

Lipopolysaccharide

MAPK:

Mitogen-activated protein kinases

p-Akt:

Protein kinase B in phosphorylated form

p-p38:

p38 in phosphorylated form

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

SVCT-2:

Sodium/vitamin C co-transporter isoform 2

TNF-α:

Tumor necrosis factor-alpha

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Funding

This work was supported by grants from Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS—process 2010/1007354) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq—process 870582/1997-5).

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Authors

Contributions

A.P.K.R and W.A.P. were responsible for the study concept and design and they prepared and wrote the manuscript. A.P.K.R., J.A.S., M.C.Q.S., A.K., E.M.S.S., T.S., A.H., C.K., and G.M. performed the experiments. L.S.F and M.F.M.R. helped in the Western blotting procedure and analysis. All authors approved the final manuscript.

Corresponding author

Correspondence to Ana Paula Konzen Riffel.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval

This study was approved by the Ethics Committee of the Universidade Federal do Rio Grande do Sul (CEUA-UFRGS #23352).

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Riffel, A.P.K., de Souza, J.A., Santos, M. et al. Co-administration of ascorbic acid and α-tocopherol modifies ascorbic acid and attenuates p38, Akt, and TNF-α expression in spinal cord of rats with neuropathic pain. Nutrire 45, 9 (2020). https://doi.org/10.1186/s41110-019-0113-6

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Keywords

  • Phosphorylated p38
  • Phosphorylated Akt
  • Chronic constriction injury
  • Sciatic nerve