Figure 1.

The in vitro studies were conducted in cultured RAW 264.7 macrophages and primary bone-marrow derive macrophages (BMDMs). Both types of cell cultures were seeded and allowed to adhere for 6 h. The culture media were then replaced with media containing sulforaphane (SFN) diluted in DMSO. These cells were either exposed to 24 h of 95% O₂ (hyperoxia) or allowed to remain at room air (21% O₂). After 24 h, different assays were conducted as described in the methods section. For the in vivo studies, male C57BL/6 mice were either exposed to > 99% O₂ or room air (21% O₂) for 72 h. During this 72 h period, blindly randomized groups of mice were given ascorbic acid (AA) or 0.9% saline (control group) intraperitoneally as indicated above. After 72 h, the mice were euthanized and their lung lavage fluid and lung tissue were harvested for assays as described in the methods section.