title: Endotoxin-induced effects on nucleotide catabolism in mouse kidney.
abstract: Extracellular adenosine 5'-triphosphate (ATP) acts as a proinflammatory mediator. Adenosine, the final product of ATP breakdown, is an anti-inflammatory compound, acting mainly on adenosine A(2A) receptors. Considering that the kidney is an organ strongly affected during systemic inflammatory responses and that ectonucleotidases are responsible for the control of extracellular nucleotide and nucleoside levels, we examined the endotoxin-induced effects on ectonucleotidases in kidney membranes of mice, and whether CGS-21680 hydrochloride (3-[4-[2-[[6-amino-9-[(2R,3R,4S,5S)-5-(ethylcarbamoyl)-3,4-dihydroxy-oxolan-2-yl]purin-2-yl]amino]ethyl]phenyl]propanoic acid), a selective adenosine A(2A) receptor agonist, antagonizes the lipopolysaccharide (LPS)-induced effects on nucleotide catabolism in kidney. Animals were injected intraperitoneally with 12 mg/kg LPS and/or 0.5mg/kg CGS-21680 or saline. Nucleotidase activities were determined in kidney membrane preparations and ATP metabolism was measured by high performance liquid chromatography (HPLC) assay. Analysis of ectonucleotidase expression was carried out by semi-quantitative semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Exposure to endotoxemia promoted an increase in ATP and p-Nitrophenyl thymidine 5'-monophosphate (p-Nph-5'-TMP) hydrolysis, and a decrease in adenosine 5'-monophosphate (AMP) hydrolysis. CGS-21680 treatment failed to reverse these changes. HPLC analysis indicated a decrease in extracellular ATP and adenosine levels in groups treated with LPS and LPS plus CGS-21680. The expression pattern of ectonucleotidases revealed an increase in Entpd3, Enpp2, and Enpp3 mRNA levels after LPS injection. These findings indicate that nucleotide and nucleoside availability in mouse kidney is altered at different stages of endotoxemia, in order to protect the integrity of this organ when exposed to systemic inflammation.
authors: Programa de Pós-Graduação em Ciências Biológicas: Bioquímica, Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600-Anexo, 90035-003, Porto Alegre, RS, Brazil.
authors: Fernanda C
authors: FC
authors: Vuaden
authors: Luiz Eduardo B
authors: LE
authors: Savio
authors: Denise B
authors: DB
authors: Ramos
authors: Emerson A
authors: EA
authors: Casali
authors: Maurício R
authors: MR
authors: Bogo
authors: Carla D
authors: CD
authors: Bonan
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D058908:Adenosine A2 Receptor Agonists / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004731:Endotoxins / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007668:Kidney / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D051379:Mice; D009708:Nucleotidases / Q000235:genetics / Q000378:metabolism; D010627:Phenethylamines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D043705:Receptor, Adenosine A2A / Q000378:metabolism
chemical_list: D058908:Adenosine A2 Receptor Agonists; D004731:Endotoxins; D008070:Lipopolysaccharides; D010627:Phenethylamines; D012333:RNA, Messenger; D043705:Receptor, Adenosine A2A; C061282:2-(4-(2-carboxyethyl)phenethylamino)-5'-N-ethylcarboxamidoadenosine; D000255:Adenosine Triphosphate; D009708:Nucleotidases; D000241:Adenosine