Fig. 2
CU06-1004 inhibits H2O2-induced reactive oxygen species generation and alleviates the inflammatory response in HBMECs. A Representative fluorescent images indicating reactive oxygen species (ROS) production in human brain microvascular epithelial cells (HBMECs). HBMECs were pretreated with 5 and 10 μg/ml CU06-1004 for 1 h, followed by incubation with 100 μM H2O2 for 2 h. Cells were then labeled with 2’, 7’-dichlorodihydrofluorescein diacetate (H2-DCFDA) to measure ROS production. B The levels of ROS were detected by fluorescence microscopy with H2-DCFDA as the fluorescent probe. Quantitative analysis was performed by measuring the fluorescence intensity relative to control cells. C CU06-1004 suppressed ROS-mediated nuclear factor-kappa-B (NF-κB) activation in HBMECs. HBMECs were pretreated with CU06-1004 for 1 h and treated with H2O2 for 6 h. D–G Quantitative analysis of intracellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and cyclooxygenase-2 (COX-2) expression levels normalized to β-actin levels. Phosphorylated NF-κB inhibitor (p-IκBα) expression levels are normalized to total IκBα levels. All expression levels were evaluated by western blotting. All data were analyzed with one-way analysis of variance, followed by Tukey’s multiple comparison test. #P < 0.05, ###P < 0.001 vs. control. **P < 0.01, ***P < 0.001 vs. H2O2. Results are presented as the mean, and error bars represent the standard error of the mean