Anandamide Protects HT22 Cells Exposed to Hydrogen Peroxide by Inhibiting CB1 Receptor-Mediated Type 2 NADPH Oxidase

Figure 3

AEA upregulated the expression of CB1 in HT22 cells. Immunofluorescence staining and western blotting were used to investigate the AEA-induced effect on CB1 protein expression in HT22 cells. 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; AM251 + AEA + H₂O₂: cells exposed to 10 μM AEA plus 10 μM CB1 antagonist AM251 at the presence of 200 μM H₂O₂ for 3 h; AM251 + H₂O₂: cells exposed to 10 μM AM251 plus 200 μM H₂O₂ for 3 h. CB1 protein (red) was expressed in HT22 cells. AEA upregulated the expression of CB1 receptor; however CB1 antagonist AM251 reversed the CB1 upregulation in HT22 cells. Nuclei were counter-stained with DAPI (blue). Results are expressed as means ± SD (𝑛=4). ^∗𝑃<0.05 versus control (no H₂O₂, no AEA, and no AM251), ^∧𝑃<0.05 versus the cells exposed to AEA plus H₂O₂. Bar = 20 μm.