The effect of glutamine on the activity of the NADPH oxidase complex from rat neutrophils was investigated. Superoxide anion (O2-) production was assessed: (1) by scintillation counting by using lucigenin, and (2) by reduction of cytochrome c over 10min. The effects of glutamine and PMA on the expression of the NADPH oxidase components p22phox, gp91phox and p47phox were also determined. Glutamine at 1 and 2mM increased O2- generation in the presence of PMA by 100% and 74% respectively, in neutrophils maintained previously for 3h in medium deprived of this amino acid. DON (6-diazo-5-oxo-l-norleucine), an inhibitor of phosphate-dependent glutaminase and thus of glutamine metabolism, caused a significant decrease in O2- production by neutrophils stimulated with PMA both in the absence (44%) and in the presence (66%) of glutamine. PMA markedly increased the expression of gp91phox, p22phox and p47phox mRNAs. Glutamine (2mM) increased the expression of these three proteins both in the absence and in the presence of PMA. We postulate that glutamine leads to O2- production in neutrophils, probably via the generation of ATP and regulation of the expression of components of NADPH oxidase.
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September 09 2002
Glutamine plays a role in superoxide production and the expression of p47phox, p22phox and gp91phox in rat neutrophils
Tania Cristina PITHON-CURI;
*Camilo Castelo Branco University and Methodist University of Piracicaba-FACEF, Rua Carolina Fonseca 584, 08230-030 Säo Paulo, Brazil
Correspondence: Dr Tania Cristina Pithon-Curi, Departamento de Fisiologia e Biofísica, Instituto de Ciências Biomédicas, Universidade de São Paulo, Av. Prof. Lineu Prestes 1524, 05508-900, Butantan, São Paulo, SP, Brazil (e-mail [email protected]).
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Adriana C. LEVADA;
Adriana C. LEVADA
*Camilo Castelo Branco University and Methodist University of Piracicaba-FACEF, Rua Carolina Fonseca 584, 08230-030 Säo Paulo, Brazil
†Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of Säo Paulo, Säo Paulo, SP, Brazil
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Lúcia R. LOPES;
Lúcia R. LOPES
‡Department of Pharmacology, Institute of Biomedical Sciences, University of Säo Paulo, Säo Paulo, SP, Brazil
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Sonia Q. DOI;
Sonia Q. DOI
§Uniformed Services University, Bethesda, MD 20814, U.S.A.
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Rui CURI
Rui CURI
†Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of Säo Paulo, Säo Paulo, SP, Brazil
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Publisher: Portland Press Ltd
Received:
May 07 2002
Revision Received:
June 13 2002
Accepted:
July 16 2002
Online ISSN: 1470-8736
Print ISSN: 0143-5221
The Biochemical Society and the Medical Research Society © 2002
2002
Clin Sci (Lond) (2002) 103 (4): 403–408.
Article history
Received:
May 07 2002
Revision Received:
June 13 2002
Accepted:
July 16 2002
Citation
Tania Cristina PITHON-CURI, Adriana C. LEVADA, Lúcia R. LOPES, Sonia Q. DOI, Rui CURI; Glutamine plays a role in superoxide production and the expression of p47phox, p22phox and gp91phox in rat neutrophils. Clin Sci (Lond) 1 October 2002; 103 (4): 403–408. doi: https://doi.org/10.1042/cs1030403
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