Figure 6

12/15-LOX metabolites induce intrinsic apoptosis in bronchial epithelia.

(A) The concentration of 12-S-HETE was estimated in serum, sputum supernatant and BAL fluid supernatant of healthy controls (Controls) or patients with atopic asthma (Atopic asthmatics). Results shown as mean ± s.e.m., significance was determined with unpaired Student's t test (*P < 0.0001). N = 40, 5 and 3 for serum, sputum and BAL fluid supernatant, respectively, for controls and N = 80, 5 and 3 for serum, sputum and BAL fluid supernatant, respectively, for atopic asthmatics. (B) The concentration of 13-S-HODE was estimated both in cytosol and culture supernatant of bronchial epithelium at IL-13 induction at the times indicated. (C) The concentration of 13-S-HODE was estimated both in cytosol and culture supernatant of bronchial epithelium with 48 hrs induction of either IL-13 alone or combination with IL-4 or TNF-α. (D) The concentration of 13-S-HODE was estimated both in culture supernatants of macrophages or fibroblasts at IL-13 induction for 24 hrs. Bronchial epithelia were incubated with 12/15-LOX metabolites rich culture supernatants of macrophages that were induced with 25 ng/ml IL-13 for 24 hrs and JC-1 staining (E) was performed and cytosolic cytochrome C levels (F) were estimated in those bronchial epithelia along with induction of different concentrations of 12-S-HETE. Carbonyl cyanide m-chlorophenylhydrazone (CCCP) treated bronchial epithelia stained with JC-1 was act as a positive control. MacroØ C.S., macrophage culture supernatant; Data are representative of two independent experiments. Results are shown as quantile box plots except 13-S-HODE and cytochrome C which were expressed in mean ± s.e.m. The line across the middle of the quantile box is the median; the ends of the box are 25th and 75th quantiles and the whiskers extending from either end of the box are the 10% and 90% quantiles. Significance is determined with unpaired Student t test (***P < 0.05).