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Am J Physiol Cell Physiol (January 15, 2003). doi:10.1152/ajpcell.00224.2002
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Submitted on May 16, 2002
Accepted on January 7, 2003

Glutamine Delays Spontaneous Apoptosis in Neutrophils

Tania C Pithon-Curi1*, Robert I Schumacher2, Jofre J Freitas3, Claudia J Lagranha3, Philip Newsholme4, Adrianne C Palanch1, Sonia Q Doi5, and Rui Curi3

1 Department of Human Performance, Methodist University of Piracicaba-FACEF and Camilo Castelo Branco University, Sao Paulo, Sao Paulo/SP, Brazil
2 Department of Biochemistry, University of Sao Paulo, Sao Paulo, Sao Paulo/SP, Brazil
3 Department of Physiology and Biophysics, University of Sao Paulo, Sao Paulo, Sao Paulo/SP, Brazil
4 Department of Biochemistry, University College Dublin, Belfield, Dublin, Ireland
5 Department of Medicine, Uniformed Services University, Bethesda, Maryland, USA

* To whom correspondence should be addressed. E-mail: tcuri{at}fisio.icb.usp.br.

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 to its absence as assessed by Hoescht 33342 staining. The level of E. coli phagocytosis in the presence of glutamine was markedly increased in comparison to the level achieved by cells cultured in the absence of glutamine. Annexin V binding to externalized phosphatidylserine was reduced in the presence of glutamine. Using sensitive fluorochrome rhodamine 123, as determined by FACS and confocal microscopy, monitored 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% and 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.




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