Anandamide Protects HT22 Cells Exposed to Hydrogen Peroxide by Inhibiting CB1 Receptor-Mediated Type 2 NADPH Oxidase
Figure 8
CB1-siRNA reversed AEA-induced protection against oxidative stress. The cells were divided into five groups, Control: cells cultured in drug-free medium; H2O2: cells exposed to 200 μM H2O2 for 3 h; AEA + H2O2: cells exposed to 10 μM AEA plus 200 μM H2O2 for 3 h; CB1-siRNA + AEA + H2O2: cells incubated with CB1-siRNA for 5 h and then exposed to 10 μM AEA plus 200 μM H2O2 for 3 h; scrambled siRNA (SC-siRNA) + AEA + H2O2: cells incubated with SC-siRNA for 5 h and then exposed to 10 μM AEA plus 200 μM H2O2 for 3 h. CB1-siRNA abolished the AEA-induced protection against 200 μM H2O2 in HT22 cells; SC-siRNA did not affect the protection. (a) CB1-siRNA significantly downregulated the expression of CB1, assessed by western blotting. (b) Cell metabolic activity, assessed by MTT (). (c) LDH release, assessed by reagent kit and spectrophotometry (). (d)–(h) The fluorescence intensity of ROS. (i) Statistical results of (d)–(h) (). Results are expressed as means ± SD, versus the control (no H2O2, no AEA, and no siRNA), versus the cells exposed to H2O2 alone, and versus the cells exposed to AEA plus H2O2.
