title: Fatty-acid-binding proteins do not protect against induced cytotoxicity in a kidney cell model.
abstract: Intracellular accumulation of fatty acids (FAs) is a well-described consequence of renal ischaemia and may lead to lethal cell injury. Fatty-acid-binding proteins (FABPs) are small cytosolic proteins with high affinity for FAs. They may protect vital cellular functions by binding to and promoting the metabolism of FAs, thereby reducing their intracellular concentration. In this study we investigated the putative cytoprotective role of FABPs in a Madin-Darby canine kidney (MDCK) cell model for renal damage. We studied the effects of transfection with cDNA encoding heart FABP, adipocyte FABP or liver FABP on cytotoxicity induced by chemical anoxia or FAs. Transfection of MDCK type II cells with these cDNA types caused a 5-20-fold increase in FABP content, but did not change the rate or extent of palmitate uptake. After 1 h of incubation with KCN, all cell types showed reduced viability and cellular ATP content and an intracellular accumulation of non-esterified FAs. High extracellular concentrations of oleate, but not palmitate, caused a markedly decreased cell viability and cellular ATP content. Oleate accumulated in non-esterified form in these cells. Simultaneous addition of glucose ameliorated the damaging effects of KCN or oleate, indicating that glycolytic ATP could substitute for uncoupled oxidative phosphorylation. No significant differences in the effects of chemical anoxia or oleate were observed between non-transfected, mock-transfected and FABP-cDNA-transfected cells. Non-esterified FA accumulation was not reduced in any of the FABP-cDNA-transfected cell lines. In conclusion, our data do not provide evidence for a cytoprotective role of FABP in this kidney cell model.
authors.affiliation: Department of Biochemistry, University Medical Center Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands. A.Zimmerman@bioch.kun.nl
authors.forename: A W
authors.initials: AW
authors.lastname: Zimmerman
authors.forename: J H
authors.initials: JH
authors.lastname: Veerkamp
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002130:Calcium Phosphates / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry* / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival; D018076:DNA, Complementary / Q000378:metabolism; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D050556:Fatty Acid-Binding Proteins; D015151:Immunoblotting; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D009363:Neoplasm Proteins; D019301:Oleic Acid / Q000378:metabolism; D019308:Palmitic Acid / Q000493:pharmacokinetics; D010957:Plasmids / Q000378:metabolism; D011190:Potassium Cyanide / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection
chemical_list: D002130:Calcium Phosphates; D002352:Carrier Proteins; D018076:DNA, Complementary; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; D019301:Oleic Acid; D019308:Palmitic Acid; C485838:calcium phosphate, monobasic, anhydrous; D000255:Adenosine Triphosphate; C020243:calcium phosphate; C485829:calcium phosphate, dibasic, anhydrous; D011190:Potassium Cyanide