Glutamine delays spontaneous apoptosis in neutrophils

• Published in: American Journal of Physiology: Cell Physiology 2003-06, Vol.284 (6), p.C1355-C1361
• Main Authors: Pithon-Curi, Tania C, Schumacher, Robert Ivan, Freitas, Jofre J. S, Lagranha, Claudia, Newsholme, Philip, Palanch, Adrianne C, Doi, Sonia Q, Curi, Rui
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis - drug effects; Cells, Cultured; Chromatin - metabolism; Diazooxonorleucine - pharmacology; Dose-Response Relationship, Drug; Enzyme Inhibitors - pharmacology; Glutamine - pharmacology; Humans; Male; Membrane Potentials - physiology; Mitochondria - metabolism; Neutrophils - cytology; Neutrophils - drug effects; Neutrophils - physiology; Phagocytosis - physiology; Phosphatidylserines - metabolism; Rats; Rats, Wistar
• ISSN: 0363-6143; 1522-1563
• DOI: 10.1152/ajpcell.00224.2002

1  Methodist University of Piracicaba, FACEF, and Camilo Castelo Branco University, São Paulo; 2  Department of Biochemistry and 3  Department of Physiology and Biophysics, University of São Paulo, São Paulo, SP, Brazil; 4  Department of Biochemistry, Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland; and 5  Uniformed Services University, Bethesda, Maryland 20814 Nuclear, mitochondrial, and plasma membrane events associated with apoptosis were investigated in rat neutrophils cultivated for 3, 24, and 48 h in the absence or presence of glutamine (0.5,   1.0, and 2.0 mM). Condensation of chromatin was reduced after 24 or 48 h of culture in the presence of glutamine compared with its absence as assessed by Hoechst 33342 staining. The level of Escherichia coli phagocytosis in the presence of glutamine was markedly increased compared with the level achieved by cells cultured in the absence of glutamine. Annexin V binding to externalized phosphatidylserine was reduced in the presence of glutamine. Sensitive fluorochrome rhodamine 123, as determined by fluorescence-activated cell sorting and confocal microscopy, was used to monitor loss of the mitochondrial transmembrane potential. In the absence of glutamine, neutrophils exhibited a marked reduction in the uptake of rhodamine 123. In the presence of 1.0 or 2.0 mM glutamine, the uptake of rhodamine was 20 or 38% higher, respectively. Similar effect was found in human neutrophils by measuring DNA fragmentation and mitochondrial transmembrane potential. Therefore, glutamine protects from events associated with triggering and executing apoptosis in both rat and human neutrophils. mitochondria; neutrophil