Neutrophil Chemotactic Activity in Bronchoalveolar Lavage Fluid of the Rats Exposed to Hyperoxia

Song, Jeong Sup; Lee, Sook Young; Moon, Wha Sik; Park, Sung Hak

Abstract

BACKGROUND
An excessive accumulation of neutrophils in lung tissue has been known to play an important role in mediating the tissue injury among the adult respiratory distress syndrome, idiopathic pulmonary fibrosis and cystic fibrosis by releasing toxic oxygen radicals and proteolytic enzymes. Therefore, it is important to understand a possible mechanism of neutrophil accumulation in lung tissue. In many species, exposure to hyperoxic stimuli can cause changes of lung tissues very similar to human adult respiratory distress syndrome and neutrophils are also functioning as the main effector cells in hyperoxic lung injury. The purpose of the present study was to examine whether neutrophils function as a key effector cell and to study the nature of possible neutrophil chemotactic factors found in bronchoalveolar lavage fluid from the hyperoxia exposed rats.
METHODS
We exposed the rats to the more than 95% oxygen for 24, 48, 60 and 72 hours and bronchoalveolar lavage(BAL) was performed. Neutrophil chemotactic activity was measured from the BAT fluid of each experimental groups. We also evaluated the molecular weight of neutrophil chemotactic factors using fast performance liquid chromatography and characterized the substances by dialyzer membrane and heat treatment.
RESULTS
1) The neutrophil proportions in bronchoalveolar lavage fluid began to rise from 48 hours after oxygen exposure, and continued to be significantly increased with exposure times. 2) Chemotactic index for neutrophils in lung lavages from rats exposed to hyperoxia was significantly higher in 48 hours group than in control group, and was significantly increased with exposure time. 3) No deaths occured until after 48 hours of exposure. However, mortality rates were increased to 33.3% in 60 hours group and 81.3% in 72 hours group. 4) Gel filtration using fast performance liquid chromatography disclosed two peaks of neutrophil chemotactic activity in molecular weight of 104,000 and 12,000 daltons. 5) Chemotactic indices of bronchoalveolar lavage fluid were significantly decreased when bronchoalveolar lavage fluid was treated with heat (56 degreesC for 30 min or 100 degreesC for 10 min) or dialyzed (dialyzer membrane molecular weight cut off: 12,000 daltons).
CONCLUSION
These results suggested that the generation of neutrophil chemotactic factor and subsequent neutrophil influx into the lungs are playing an important roles in hyperoxia-induced acute lung injury. Neutrophil chemotactic factor in the lung lavage fluids consisted of several distinct components having different molecular weight and different physical characteristics.