Figure 6: Flaxseed blunts increased Lung Fibrosis and Profibrogenic Cytokine Levels induced by Exposure to Hyperoxia, Radiation, or Double-Hit Combination Challenge.Separate cohorts of mice (n=5/group) were fed 0% or 10% FS diet and exposed to 100% O2 for 8 hours only (O2), 0.25 Gy total body ionizing gamma irradiation(IR) only, or a double-hit combination of both challenges (O2+IR) followed by intermittent normoxia (21% pO2) for repeated cycles 3 times a week for 2 weeks. Mice were sacrificed after 2 weeks of challenge exposure. Lung tissue was harvested and evaluated for hydroxyproline content, a measure of fibrosis (Panel A). Changes in lung tissue mRNA levels of profibrogenic TGFß1 were determined quantitative real-time PCR analysis (Panel B). Analysis was performed in duplicate and gene expression normalized to 18S ribosomal RNA. Levels of active TGFβ1 were determined in mouse BALF using ELISA (Panel C). All data is represented as mean ± SEM. *p< 0.05 for O2, IR, and O2+IR versus respective untreated control mice in each diet cohort. #p< 0.05 for 0% FS fed mice exposed to O2, IR, and O2+IR versus respective 10% FS fed mice exposed to O2, IR, and O2+IR.