RT Journal Article
SR Electronic
T1 Effects of anaerobiosis upon morphology and energy metabolism of alveolar macrophages cultured in gas phase
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP 1015
OP 1022
DO 10.1183/09031936.93.03091015
VO 3
IS 9
A1 M Cazin
A1 D Paluszezak
A1 A Bianchi
A1 JC Cazin
A1 C Aerts
A1 C Voisin
YR 1990
UL http://erj.ersjournals.com/content/3/9/1015.abstract
AB Metabolic and morphological effects of anoxia were studied in alveolar macrophages obtained by lung lavage from guinea-pigs by means of an original method of cell culture allowing direct contact with air without interposition of liquid medium. After selection by glass adherence, alveolar cells were layered on a porous membrane applied to the surface of a reservoir filled with nutrient medium. Alveolar macrophages were then cultured in gas phase under either aerobic or anaerobic conditions for 24, 48 and 72 h. Cellular adenosine triphosphate (ATP) content, an indicator of cell vitality, significantly decreased by 68 and 88% after 48 and 72 h of exposure to anaerobic environment, respectively. Significant increases in lactate production (68% at 24 h) and in glucose uptake (125% at 24 h), evidence of marked glycolytic activity, occurred before these falls in intracellular ATP and parallel decreases in culture medium pyruvate level (76 and 85% at 48 and 72 h, respectively). The shift of energy metabolism resulted in cell death after 72 h, as noted by morphological degeneration and decreased cellular ATP content. Twenty-four hour re-exposure to normoxic atmosphere showed that recovery was possible when duration of anaerobiosis did not exceed 48 h. This reversibility in anoxic cell injury has been related to plasma membrane integrity. The results of these studies indicate that alveolar macrophage resistance to anaerobiosis is limited as ATP content falls and morphological degeneration occurs after 48 h. This novel approach of anaerobic effects at the cell level should be adaptable to investigations of activity and, in particular, the mechanisms of metabolic activity of antianoxic drugs.