Rossetto Isabela Maria Urra, Cagnon Valéria Helena Alves, Kido Larissa Akemi, Lizarte Neto Fermino Sanches, Tirapelli Luís Fernando, Tirapelli Daniela Pretti da Cunha, de Almeida Chuffa Luiz Gustavo, Martinez Francisco Eduardo, Martinez Marcelo
Department of Structural and Functional Biology, University of Campinas (UNICAMP), 255 Monteiro Lobato St, Campinas, SP 13083-862, Brazil.
Department of Food and Nutrition, University of Campinas (UNICAMP), 80 Monteiro Lobato St, Campinas, SP 13083-862, Brazil.
Toxicol Res (Camb). 2021 Jul 24;10(4):835-849. doi: 10.1093/toxres/tfab067. eCollection 2021 Aug.
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 ( = 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.
咖啡因的摄入能够通过腺苷能系统发挥作用，干扰与炎症机制相关的细胞过程。本研究旨在识别智利大学嗜酒（UChB）大鼠在摄入乙醇和咖啡因后，其小脑腺苷能系统和炎症过程的相关变化。5月龄雄性UChB大鼠和Wistar大鼠被分为以下几组（每组n = 15）：（i）对照组（Wistar大鼠饮用清水）；（ii）乙醇组（UChB大鼠饮用10%乙醇溶液）；（iii）乙醇+咖啡因组（UChB大鼠饮用添加了3 g/L咖啡因的10%乙醇溶液）。收集小脑组织并进行处理，用于免疫组织化学、逆转录聚合酶链反应（RT-PCR）和蛋白质印迹技术，以检测腺苷能受体A1和A2a以及炎症标志物，包括核因子κB（NFκB）、Toll样受体4（TLR4）、Toll样受体2（TLR2）、髓样分化因子88（MyD88）、肿瘤坏死因子-α（TNF-α）、环氧化酶-2（COX-2）、诱导型一氧化氮合酶（iNOS）和小胶质细胞标志物离子钙结合衔接分子1（Iba-1）。结果显示，乙醇和咖啡因的摄入分别改变了小脑组织中腺苷能受体和炎症标志物的免疫定位。A2a受体在乙醇组中过度表达，在神经胶质细胞中明显可见。乙醇组中NFκB和TLR4的蛋白质水平升高，在伯格曼胶质细胞和浦肯野细胞中表现明显。咖啡因将这些标志物的表达降低至与对照组相似的水平。乙醇组中A1基因上调，但其蛋白质水平未上调，提示存在转录后干扰。总之，咖啡因似乎通过调节A1和A2a减轻了UChB大鼠小脑中乙醇诱导的炎症，在乙醇摄入的慢性情况下发挥神经保护作用。
