Comparative toxicity of fatty acids on a macrophage cell line (J774).
In the present study, the cytotoxicity of palmitic, stearic, oleic, linoleic, arachidonic, docosahexaenoic and eicosapentaenoic acids on a macrophage cell line (J774) was investigated. The induction of toxicity was investigated by changes in cell size, granularity, membrane integrity, DNA fragmentation and phosphatidylserine externalization by using flow cytometry. Fluorescence microscopy was used to determine the type of cell death (Acridine Orange/ethidium bromide assay). The possible mechanisms involved were examined by measuring mitochondrial depolarization, lipid accumulation and PPARgamma (peroxisome-proliferator-activated receptor gamma) activation. The results demonstrate that fatty acids induce apoptosis and necrosis of J774 cells. At high concentrations, fatty acids cause macrophage death mainly by necrosis. The cytotoxicity of the fatty acids was not strictly related to the number of double bonds in the molecules: palmitic acid>docosahexaenoic acid>stearic acid=eicosapentaenoic acid=arachidonic acid>oleic acid>linoleic acid. The induction of cell death did not involve PPARgamma activation. The mechanisms of fatty acids to induce cell death involved changes in mitochondrial transmembrane potential and intracellular neutral lipid accumulation. Fatty acids poorly incorporated into triacylglycerol had the highest toxicity.
Martins de Lima
Thais
T
Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, CEP 05508-900, São Paulo, Brazil. thais@fisio.icb.usp.br
Cury-Boaventura
Maria Fernanda
MF
Giannocco
Gisele
G
Nunes
Maria Tereza
MT
Curi
Rui
R
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D003124:Colorimetry; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005227:Fatty Acids / Q000493:pharmacokinetics / Q000633:toxicity*; D050356:Lipid Metabolism / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D009336:Necrosis; D047495:PPAR gamma / Q000378:metabolism; D014280:Triglycerides / Q000378:metabolism
D005227:Fatty Acids; D047495:PPAR gamma; D014280:Triglycerides