Table 2.

Structural and functional alterations of A549 cells when treated with pure recombinant PA-LOX. Human lung epithelial cells A549 were seeded in 6-well plates and grown to pre-confluence in the presence of FCS. Monolayers were gently washed with serum free DMEM and 2 ml of this medium containing 250 μg/ml of pure PA-LOX was added. After 24 h the supernatant was removed, adherent cells were gently washed with DPBS, scraped off and reconstituted in 2 ml DMEM. Detached and adherent cells were counted and trypan blue staining was performed with small aliquots of the two cell suspensions. From the remaining cells, the lipids were extracted, hydrolyzed under alkaline conditions and the OH-PUFA/PUFA-ratio was quantified as described in Materials and methods. The experiment was carried out in triplicate and means ± SD are given in the table.

Parameter	Control incubation	PA-LOX incubation	Significance (p)
Cells in supernatant (%)	2.1 ± 0.7	7.1 ± 1.6	< 0.001
Dead cell in supernatant (%)	62.6 ± 5.7	81.1 ± 4.0	0.01
Adherent dead cells (%)	19.4 ± 7.1	68.8 ± 2.4	< 0.001
OH-PUFA/PUFA ratio (%)	0.17 ± 0.09	3.6 ± 0.4	< 0.001