FIGURE 5.

Inhibition of TRAF6 attenuated LPS-mediated myocardial injury. The H9C2 cardiomyocytes were pretreated with C25-140 (5 μM, an inhibitor of TRAF6) for 2 h and then treated with LPS (10 μg/ml) and Deh (10 μg/ml) for 24 h (A, B) CCK8, and BrdU assay were taken to detect the cell viability of each group. The rate of BrdU positive cells (red)/nucleus (blue) was calculated. (C) Western blot was performed for the detection of Bax, Caspase3 and Bcl2 expressions in cardiomyocytes. (D, E) ELISA method was employed to evaluate IL-1β and TNFα levels in the culture medium of each group. (F, G) The SOD and GSH-PX detection kits were applied to detect SOD (G) and GSH-PX (H) levels in the culture medium of each group. (H, I). The ROS level in H9C2 cells were evaluated using the DCFDA/H2DCFDA - Cellular ROS Assay Kit. (J, K). The protein level of iNOS, Nrf2/HO-1, TRAF6, and p-NF-κB in the whole cell or nucleus of heart was detected by western blot. nsP> 0.05, **p < 0.01, ***p < 0.001. N = 3.