Figure 3.

Triptolide treatment decreased the generation of reactive oxygen species (ROS) by activating NRF2/HO-1 signaling cascades. RAW 264.7 macrophages were treated with 1,000 ng/ml LPS with 0–40 nM triptolide for 24 h. (A, B) Flow cytometry was performed to determine the generation of ROS in intestinal macrophages in DSS-induced mice colitis. (C, D) The DCFH-DA probe was used to measure the ROS generation in RAW264.7 cells by flow cytometry. (E) Immunofluorescence assays were performed to detect the cytoplasmic and nuclear NRF2 and HO-1 levels in RAW 264.7 macrophages. (F) RT-qPCR was used to detect the mRNA expression of MPO, Gss, Gclc, and Gclm in RAW 264.7 macrophages. (G, H) The ROS inhibition effect of triptolide was significantly reversed by the ML385 in LPS-activated macrophages. β-actin or glyceraldehyde 3-phosphate dehydrogenase served as an internal control. Data are shown as the mean ± SD of at least three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001; and ns means no significance. ## means the statistical difference between the results of the LPS group and the mock group was P < 0.01.