2.6. Cell Culture and Treatment

The macrophage cell line RAW264.7 was obtained from ATCC (Manassas, VA, USA), which was cultured in Roswell Park Memorial Institute- (RPMI-) 1640 medium containing 10% fetal bovine serum (FBS). The RAW264.7 macrophages were treated with 100 ng/ml of LPS for 24 h at 37°C to establish cellular ARDS models [27]. To study the potential effects of pomiferin on oxidative stress and inflammatory response in macrophages, pomiferin (50 μM) was used to treat the RAW264.7 macrophages stimulated with LPS. To active AKT, 30 μM of 3-deoxysappanchalcone (3-DE, #HY-N1745A, Shanghai, MCE, China) was used to pretreat RAW264.7 macrophages for 30 min [28].