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

Figure 10

Nox inhibitor did not induce a more significant reduction of Nox2 expression than AEA alone. 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; Apocynin + AEA + H₂O₂: cells exposed to 10 μM AEA plus 50 μM Nox inhibitor AM251 in the presence of 200 μM H₂O₂ for 3 h; Apocynin + AEA + H₂O₂: cells exposed to 50 μM apocynin plus 10 μM AEA in the presence of 200 μM H₂O₂ for 3 h. Nox2 protein expression (a) was evaluated by western blotting (𝑛=4). (b) Cell metabolic activity and (c) LDH release were determined by MTT (𝑛=8) and reagent kit (𝑛=6), respectively. Results are expressed as means ± S.D. ^∗𝑃<0.05 versus the control (no H₂O₂, no AEA, and no apocynin), ^#𝑃<0.05 versus the cells exposed to H₂O₂ alone, n.s.: no significance.

(a)

(b)

(c)