Creatine supplementation increases postnatal growth and strength and prevents overexpression of pro-inflammatory interleukin 6 in the hippocampus in an experimental model of cerebral palsy

ABSTRACT

Objectives: The aim of this study was thus to evaluate the effect of Cr supplementation on morphological changes and expression of pro-inflammatory cytokines in the hippocampus and on developmental parameters. Methods: Male Wistar rat pups were submitted to an experimental model of CP. Cr was administered via gavage from the 21st to the 28th postnatal day, and in water after the 28th, until the end of the experiment. Body weight (BW), food consumption (FC), muscle strength, and locomotion were evaluated. Expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) were assessed in the hippocampus by quantitative real-time polymerase chain reaction. Iba1 immunoreactivity was assessed by immunocytochemistry in the hippocampal hilus. Results: Experimental CP caused increased density and activation of microglial cells, and overexpression of IL-6. The rats with CP also presented abnormal BW development and impairment of strength and locomotion. Cr supplementation was able to reverse the overexpression of IL-6 in the hippocampus and mitigate the impairments observed in BW, strength, and locomotion. Discussion: Future studies should evaluate other neurobiological characteristics, including changes in neural precursor cells and other cytokines, both pro- and anti-inflammatory.

KEYWORDS:

• Cerebral palsy
• growth and development
• inflammation
• microglia
• cytokines
• hippocampus
• locomotion
• muscle strength

Acknowledgements

We also would like to thank the ‘Central Multiusuário do Departamento de Física (DF) da Universidade Federal de Pernambuco (UFPE)’ for making available the reflected-light optical microscope for the analyses. Figures 1 and 8 were created with BioRender.com.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data that support the findings of this study are available from the corresponding author, A.E.T, upon reasonable request.

Author contributions

H.J.C.B.G., B.J.R.C.-d.-S., E.E.M.V, and E.R.S conducted the experiments. H.J.C.B.G. and B.J.R.C.-d.-S. peformed the immunostaining and microscopical analyses. A.R. and L.T. carried out the real-time PCR analyses. H.J.C.B.G. analyzed the data, prepared the graphs and tables, and wrote the manuscript. O.G.-Q and A.E.T contributed to discussion. A.E.T. and R.M.-d.-C. conceptualized the study and supervised the work. All authors have read and agreed to the published version of the manuscript.

Additional information

Funding

This study was funded in part by the ‘Tecnológico Nacional de México’ (15318.22-PD) – Mexico; ‘Instituto Mexicano del Seguro Social’ (R-2019-1602-015); ‘Fundação de Amparo à Ciência e Tecnologia de Pernambuco’ (FACEPE, #0176-4.05/19; #0989-4.05/22 and #1471-4.05/22) – Brazil; ‘Coordenação de Aperfeiçoamento de Pessoal de Nível Superior’ (CAPES) (Financial code – 001) – Brazil; ‘Conselho Nacional de Desenvolvimento Científico e Tecnológico’ (CNPq) (#402426/2021-5) – Brazil.

Notes on contributors

Henrique J. C. B. Gouveia

Henrique J. C. B. Gouveia is a postdoctoral student at the Postgraduate Program in Nutrition, Federal University of Pernambuco, Recife, Brazil. His work is developed in the field of neuroscience, nutrition and phenotypic plasticity.

Raul Manhães-de-Castro

Raul Manhães-de-Castro is an Emeritus Professor at Federal University of Pernambuco, Recife, Brazil. His work is developed in the field of neuroscience, nutrition and phenotypic plasticity.

Bárbara J. R. Costa-de-Santana

Bárbara J. R. Costa-de-Santana is a doctoral student at the ‘Graduate Program in Neuropsychiatry and Behavioral Sciences’, Federal University of Pernambuco, Recife, Brazil. She carries out her work in the field of neuroscience, nutrition, and phenotypic plasticity.

Emanuel Ewerton M. Vasconcelos

Emanuel Ewerton M. Vasconcelos is a scientific initiation student and a graduate student in dentistry at the ‘Department of Clinical and Preventive Dentistry and Department of Prosthodontics and Oral and Facial Surgery’, Federal University of Pernambuco, Recife, Brazil.

Eliesly Roberto Silva

Eliesly Roberto Silva is a scientific initiation student and a graduate student in dentistry at the ‘Department of Clinical and Preventive Dentistry and Department of Prosthodontics and Oral and Facial Surgery’, Federal University of Pernambuco, Recife, Brazil.

Angélica Roque

Angélica Roque is an assistant researcher in the laboratory of Dr. Torner at Centro de Investigación Biomética de Michoacán’, Instituto Mexicano del Seguro Social, Morelia, Michoacán, México. She works with animal models of early life stress and inflammatory markers in the brain.

Luz Torner

Luz Torner is a Titular A Researcher at ‘Centro de Investigación Biomédica de Michoacán’, Instituto Mexicano del Seguro Social, Morelia, Michoacán, México. She carries out her studies on mechanisms that operate in the brain during stress exposure to produce the vulnerability of the individual to develop psychiatric diseases.

Omar Guzmán-Quevedo

Omar Guzmán-Quevedo is a Professor at ‘Higher Technological Institute of Tacambaro’, Tacambaro, Michoacán, Mexico. He carries out his work in the field of neuroscience, nutrition, and phenotypic plasticity, focusing on the study of the pathogenesis and therapy of metabolic disorders.

Ana E. Toscano

Ana E. Toscano is a Professor at ‘Nursing Unit, Vitória Academic Center’, and at ‘Graduate Program in Neuropsychiatry and Behavioral Sciences (Posneuro)’, Federal University of Pernambuco. Her work is carried out in the field of neuroscience, nutrition, cerebral palsy, and phenotypic plasticity.