title: The effects of lipopolysaccharide-induced reactive oxygen species were blunted by calcium oxalate in renal tubular epithelial cells.
abstract: OBJECTIVE
  Previously we demonstrated that calcium oxalate (CaOx) in LLC-PK1 cells and oxalate in MDCK cells induce tubular damage and greater glycosaminoglycan synthesis. We test the hypothesis that reactive oxygen species (ROS) and prostaglandins mediate these effects.
  
  
  METHODS
  LLC-PK1 and MDCK cells were exposed to graded concentrations of CaOx, oxalate or both. Glycosaminoglycan synthesis was analyzed through metabolic labeling and gel electrophoresis. Cell permeability and lipid peroxidation were assessed by lactate dehydrogenase release and malondialdehyde levels. Hydrogen peroxide and superoxide anion were analyzed using 2',7'-dichlorofluorescein and luminol. Cyclooxygenase-2 expression and prostaglandin E2 production were assessed by RT-PCR and ELISA, respectively.
  
  
  RESULTS
  In LLC-PK1 cells exposed to CaOx, we observed increased cell permeability, no induction of ROS or lipid peroxidation, inability to produce lipopolysaccharide-induced ROS and increases in prostaglandin E2. Indomethacin used alone increased glycosaminoglycan synthesis but did not potentiate CaOx-induced effects. In MDCK cells exposed to oxalate we observed increased cell permeability, ROS production only at higher concentrations and inability to produce lipopolysaccharide-induced ROS. Indomethacin alone had no effect but increased oxalate-induced glycosaminoglycan synthesis.
  
  
  CONCLUSIONS
  Prostaglandins modulate endogenous production of glycosaminoglycans in LLC-PK1 cells, as well as regulate oxalate-induced glycosaminoglycan synthesis in MDCK cells. Rather than increasing, CaOx and oxalate blunted lipopolysaccharide-induced ROS production. We could speculate that patients with recurrent nephrolithiasis may lose antimicrobial protection induced by ROS during infections.
authors.affiliation: Departamento de Medicina, Disciplina de Nefrologia, Universidade Federal de São Paulo (UNIFESP), São Paulo, Brasil.
authors.forename: F T
authors.initials: FT
authors.lastname: Borges
authors.forename: A S
authors.initials: AS
authors.lastname: Garofalo
authors.forename: M A
authors.initials: MA
authors.lastname: Dalboni
authors.forename: N P
authors.initials: NP
authors.lastname: Abreu
authors.forename: Y M
authors.initials: YM
authors.lastname: Michelacci
authors.forename: N
authors.initials: N
authors.lastname: Schor
mesh_terms: D000818:Animals; D002129:Calcium Oxalate / Q000494:pharmacology*; D002460:Cell Line; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007686:Kidney Tubules, Distal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018374:LLC-PK1 Cells; D008070:Lipopolysaccharides / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D013552:Swine
chemical_list: D008070:Lipopolysaccharides; D017382:Reactive Oxygen Species; D002129:Calcium Oxalate