Figure 3.
Inhibition of CYP4A with HET0016 protects H9c2 cells from AGEs-induced oxidative stress. (A) Protein expression of CYP4A, n = 3. (B–D) mRNA expressions of CYP4a1, CYP4a2, CYP4a3, n = 3. (E) Level of intracellular ROS generation. (F) A summary of flow cytometry analyses of cells stained with 2,7-dichlorodihydrofluoresceindiacetate (DCFH-DA), n = 3. (G) LDH activity in the medium, n = 6. (H) Content of MDA, n = 6. (I) Protein expression of NOX2, n = 3. (J) Activity of SOD, n = 6. H9c2 cells at a density of 1 × 105 cells per well were seeded on six-well plates and treated at the time of 90% confluence. HET0016 or vehicle in fetal bovine serum (PBS)-free media (2 μM) was added 1 h before exposure to 10 μM AGEs or bovine serum albumin (BSA) for 24 h. Con, control. Compared with the Con group, ++p < 0.01, +++p < 0.001; compared with the AGEs group, *p < 0.05, **p < 0.01 (one-way ANOVA with Tukey post hoc).