Effects of caffeine on behavioral and inflammatory changes elicited by copper in zebrafish larvae: Role of adenosine receptors.
This study investigated the effects of caffeine in the behavioral and inflammatory alterations caused by copper in zebrafish larvae, attempting to correlate these changes with the modulation of adenosine receptors. To perform a survival curve, 7dpf larvae were exposed to 10μM CuSO4, combined to different concentrations of caffeine (100μM, 500μM and 1mM) for up to 24h. The treatment with copper showed lower survival rates only when combined with 500μM and 1mM of caffeine. We selected 4 and 24h as treatment time-points. The behavior evaluation was done by analyzing the traveled distance, the number of entries in the center, and the length of permanence in the center and the periphery of the well. The exposure to 10μM CuSO4plus 500μM caffeine at 4 and 24h changed the behavioral parameters. To study the inflammatory effects of caffeine, we assessed the PGE2levels by using UHPLC-MS/MS, and TNF, COX-2, IL-6 and IL-10 gene expression by RT-qPCR. The expression of adenosine receptors was also evaluated with RT-qPCR. When combined to copper, caffeine altered inflammatory markers depending on the time of exposure. Adenosine receptors expression was significantly increased, especially after 4h exposure to copper and caffeine together or separately. Our results demonstrated that caffeine enhances the inflammation induced by copper by decreasing animal survival, altering inflammatory markers and promoting behavioral changes in zebrafish larvae. We also conclude that alterations in adenosine receptors are related to those effects.
Programa de Pós-Graduação em Medicina e Ciências da Saúde, PUCRS, Avenida Ipiranga, 6690, 90619-900 Porto Alegre, RS, Brazil; Laboratório de Farmacologia Aplicada, PUCRS, Avenida Ipiranga, 6681, Partenon, 90619-900 Porto Alegre, RS, Brazil.
Fernanda Fernandes
FF
Cruz
Instituto de Toxicologia e Farmacologia, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil.
Carlos Eduardo
CE
Leite
Programa de Pós-Graduação em Medicina e Ciências da Saúde, PUCRS, Avenida Ipiranga, 6690, 90619-900 Porto Alegre, RS, Brazil; Laboratório de Genômica e Biologia Molecular, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil.
Luiza Wilges
LW
Kist
Programa de Pós-Graduação em Medicina e Ciências da Saúde, PUCRS, Avenida Ipiranga, 6690, 90619-900 Porto Alegre, RS, Brazil; Laboratório de Genômica e Biologia Molecular, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil.
Giovanna Medeiros
GM
de Oliveira
Faculdade de Biociências, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil; Laboratório de Genômica e Biologia Molecular, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil.
Maurício Reis
MR
Bogo
Faculdade de Biociências, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil.
Carla Denise
CD
Bonan
Instituto de Toxicologia e Farmacologia, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil; Faculdade de Odontologia, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil.
Maria Martha
MM
Campos
Programa de Pós-Graduação em Medicina e Ciências da Saúde, PUCRS, Avenida Ipiranga, 6690, 90619-900 Porto Alegre, RS, Brazil; Laboratório de Farmacologia Aplicada, PUCRS, Avenida Ipiranga, 6681, Partenon, 90619-900 Porto Alegre, RS, Brazil; Faculdade de Farmácia, PUCRS, Avenida Ipiranga, 6681, 90619-900 Porto Alegre, RS, Brazil. Electronic address: fernanda.morrone@pucrs.br.
Fernanda Bueno
FB
Morrone
D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects; D015415:Biomarkers / Q000378:metabolism; D002110:Caffeine / Q000009:adverse effects* / Q000819:agonists / Q000037:antagonists & inhibitors; D003300:Copper / Q000819:agonists / Q000737:chemistry / Q000633:toxicity*; D019327:Copper Sulfate / Q000008:administration & dosage; D015232:Dinoprostone / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D018836:Inflammation Mediators / Q000819:agonists / Q000378:metabolism; D007814:Larva / Q000187:drug effects* / Q000254:growth & development / Q000276:immunology / Q000378:metabolism; D009994:Osmolar Concentration; D058906:Purinergic P1 Receptor Agonists / Q000737:chemistry / Q000633:toxicity; D058915:Purinergic P1 Receptor Antagonists / Q000009:adverse effects* / Q000737:chemistry; D018047:Receptors, Purinergic P1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016019:Survival Analysis; D014874:Water Pollutants, Chemical / Q000819:agonists / Q000037:antagonists & inhibitors / Q000633:toxicity*; D015027:Zebrafish / Q000254:growth & development / Q000276:immunology / Q000502:physiology*; D029961:Zebrafish Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
D015415:Biomarkers; D018836:Inflammation Mediators; D058906:Purinergic P1 Receptor Agonists; D058915:Purinergic P1 Receptor Antagonists; D018047:Receptors, Purinergic P1; D014874:Water Pollutants, Chemical; D029961:Zebrafish Proteins; D002110:Caffeine; D003300:Copper; D015232:Dinoprostone; D019327:Copper Sulfate
Adenosine receptors; Caffeine; Copper; Inflammation; Purinergic system; Zebrafish