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. 2018 Jun 26;9:675. doi: 10.3389/fphar.2018.00675

FIGURE 6.

FIGURE 6

Involvement of α7nAChR and PI3K/Akt/mTOR pathway in the sinomenine (SIN)-induced VIP expression in PC-12 cells. (A,B) Cells were pre-treated with H-89, GF109203X, LY-294002 or U0126 (10 μM). After 30 min, cells were exposed to vehicle, nicotine (NIC) or SIN for 24 h. The mRNA and protein level of VIP were measured by quantitative PCR assay and ELISA respectively. (C) The expression of Akt and p-Akt were detected by western blot. (D) Cells were pre-treated with LY-294002 or α-bungarotoxin (α-BTX, 0.1 μM). After 30 min, they were exposed to vehicle, NIC (1 μM) or SIN (0.3 mM) for 24 h. The expression of Akt and p-Akt were detected by western blot. (E) Cells were pre-treated with rapamycin (Rapa, 10 nm). After 30 min, they were exposed to vehicle, NIC (1 μM) or SIN (0.3 mM) for 24 h. The expression of RPS6 and p-RPS6 were detected by western blot. Data were expressed as means ± SEM of three independent experiments. ∗P < 0.05 and ∗∗P < 0.01 versus Control; $P < 0.05 and $$P < 0.01 versus NIC; &P < 0.05 versus SIN.