Caffeine consumption attenuates ethanol-induced inflammation through the regulation of adenosinergic receptors in the UChB rats cerebellum

Isabela Maria Urra Rossetto; Valéria Helena Alves Cagnon; Larissa Akemi Kido; Fermino Sanches Lizarte Neto; Luís Fernando Tirapelli; Daniela Pretti da Cunha Tirapelli; Luiz Gustavo de Almeida Chuffa; Francisco Eduardo Martinez; Marcelo Martinez

doi:10.1093/toxres/tfab067

Caffeine consumption attenuates ethanol-induced inflammation through the regulation of adenosinergic receptors in the UChB rats cerebellum

Toxicol Res (Camb). 2021 Jul 24;10(4):835-849. doi: 10.1093/toxres/tfab067. eCollection 2021 Aug.

Authors

Isabela Maria Urra Rossetto¹, Valéria Helena Alves Cagnon¹, Larissa Akemi Kido², Fermino Sanches Lizarte Neto³, Luís Fernando Tirapelli³, Daniela Pretti da Cunha Tirapelli³, Luiz Gustavo de Almeida Chuffa⁴, Francisco Eduardo Martinez⁴, Marcelo Martinez⁵

Affiliations

¹ Department of Structural and Functional Biology, University of Campinas (UNICAMP), 255 Monteiro Lobato St, Campinas, SP 13083-862, Brazil.
² Department of Food and Nutrition, University of Campinas (UNICAMP), 80 Monteiro Lobato St, Campinas, SP 13083-862, Brazil.
³ Department of Surgery and Anatomy, University of São Paulo (USP), 3900 Bandeirantes Ave, Ribeirão Preto, SP 14049-900, Brazil.
⁴ Department of Structural and Functional Biology, State University of São Paulo (UNESP), 250 Prof. Dr. Antônio Celso Wagner Zanin St, Botucatu, SP 18618-689, Brazil.
⁵ Department of Morphology and Pathology, Federal University of São Carlos (UFSCar), 13571 Biblioteca Comunitária Ave, São Carlos, SP 13565-905, Brazil.

Abstract

Caffeine consumption is able to interfere in cellular processes related to inflammatory mechanisms by acting through the adenosinergic system. This study aimed to recognize alterations related to adenosinergic system and inflammatory process in the cerebellum of University of Chile Bibulous (UChB) rats after the consumption of ethanol and caffeine. UChB and Wistar rats, males at 5 months old, were divided into the groups (n = 15/group): (i) Control (Wistar rats receiving water); (ii) Ethanol group (UChB rats receiving ethanol solution at 10%) and (iii) Ethanol+caffeine group (UChB rats receiving ethanol solution at 10% added of 3 g/L of caffeine). The cerebellar tissue was collected and processed for immunohistochemistry, Reverse transcription polymerase chain reaction (RT-PCR) and western blotting techniques for the adenosinergic receptors A1 and A2a and inflammatory markers, including Nuclear factor kappa B (NFkB), TLR4, TLR2, MyD88, TNF-α, COX-2, iNOS and microglial marker Iba-1. Results showed ethanol and caffeine consumption differentially altering the immunolocalization of adenosinergic receptors and inflammatory markers in the cerebellar tissue. The A2a receptor was overexpressed in the Ethanol group and was evident in the glial cells. The Ethanol group had increased protein levels for NFκB and TLR4, expressively in Bergmann glia and Purkinje cells. Caffeine reduced the expression of these markers to levels similar to those found in the Control group. The A1 gene was upregulated the Ethanol group, but not its protein levels, suggesting post-transcriptional interference. In conclusion, caffeine seems to attenuate ethanol-induced inflammation in the cerebellum of UChB rats through the A1 and A2a modulation, playing a neuroprotective role in the chronic context of ethanol consumption.

Keywords: UChB rats; adenosinergic receptor; caffeine; cerebellum; ethanol; inflammation.