Percentage of phagocytosis, production of O2·−, H2O2 and NO, and antioxidant enzyme activities of rat neutrophils in culture
Article first published online: 21 DEC 1998
Copyright © 1998 John Wiley & Sons, Ltd.
Cell Biochemistry and Function
Volume 16, Issue 1, pages 43–49, March 1998
How to Cite
Curi, T. C. P., De Melo, M. P., Palanch, A. C., Miyasaka, C. K. and Curi, R. (1998), Percentage of phagocytosis, production of O2·−, H2O2 and NO, and antioxidant enzyme activities of rat neutrophils in culture. Cell Biochem. Funct., 16: 43–49. doi: 10.1002/(SICI)1099-0844(199803)16:1<43::AID-CBF761>3.0.CO;2-5
- Issue published online: 21 DEC 1998
- Article first published online: 21 DEC 1998
- Manuscript Accepted: 7 JUN 1997
- Manuscript Received: 17 FEB 1997
- reactive species of oxygen;
- superoxide dismutase;
- catalase and peroxidase
Changes in the integrity, ultrastructure, phagocytosis capacity, and production of H2O2, O2· −and NO2− were evaluated in cultured neutrophils. The activities of the antioxidant enzymes (catalase—CAT, superoxide dismutase—SOD and glutathione-dependent peroxidase—GSH-Px) were measured under similar conditions. The integrity of the cells remained unchanged up to 18 h. After 24 h, the number of viable cells in culture dropped by 16 per cent. The percentage of viable cells in culture was of 72 per cent even after 72 h. An ultrastructural analysis of the cells was carried out after 3, 6, 12, 24, 48, and 72 h in culture. Neutrophils started developing morphologic changes after 24 h: decreased cell volume, abundant vacuoles (mainly around the nucleus), and also the presence of autophagic vacuoles. This period was then chosen for the study of neutrophil function and antioxidant enzyme activities. Neutrophils cultured for 24 h presented reduced phagocytosis capacity. The rates of production of H2O2 and O2· − remained unchanged after 24 h in culture. Concomitantly, these cells were also able to produce NO in significant amounts. The production of O2·− in response to PMA stimulus was lowered in 24-h cultured cells. Possibly, the production of oxygen and nitrogen reactive species accomplished with a decrease in the activities of CAT and GSH-Px play a key role for the process of apoptosis which takes place in neutrophils under these conditions. © 1998 John Wiley & Sons, Ltd.