Figure 4

Effects of betulin on anti-inflammatory and antioxidant reactions in LPS-stimulated macrophages. RAW264.7 cells were plated in 12-well plates, preincubated with betulin (9, 18 and 36 μM) for 1 h, and then challenged with LPS (500 ng/ml) for 18 h. (a) The expression of proinflammatory genes (iNOS and COX-2) and antioxidant enzymes (HO-1 and NQO1) were detected by quantitative real-time PCR. (b and c) The cells were incubated with 50 μM of DCFH-DA for 40 min, and DCF fluorescence intensities were detected by flow cytometry. (d) The cells were washed with PBS and incubated with JC-1 (10 μg/ml) at 37 °C in the dark for 30 min. JC-1 fluorescence was quantified through flow cytometry. *P⩽0.05, **P⩽0.01 versus the LPS group