Research Paper Volume 13, Issue 7 pp 9542—9565

Gastrodin ameliorates learning and memory impairment in rats with vascular dementia by promoting autophagy flux via inhibition of the Ca2+/CaMKII signal pathway

GAS ameliorated CoCl2-induced autophagic flux inhibition in HT22 cells. (A) Levels of LC3, p62, and p-p62 (Ser349) in HT22 cells treated with CQ were assessed, with or without GAS (200 μM) for 24 h (n = 3). (B) Levels of LC3, p62, and phosphorylated p62 (Ser349) in HT22 cells treated with 3-BDO were assessed with or without GAS (200 μM) for 24 h (n = 3). (C) Detection of p62 siRNA transfection efficiency by western blotting. (D–F) Levels of LAMP-2, p62, and phosphorylated p62 (Ser349) in HT22 cells transfected with nonspecific or p62 siRNA were evaluated with or without GAS (200 μM) for 24 h (n = 3). The experimental results were normalized to β-actin levels and are shown as fold changes relative to control cells. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments. ##P*P2. GAS, gastrodin; CoCl2, cobalt chloride; LC3, microtubule-associated protein 1 light chain 3; CQ, chloroquine.

Figure 4. GAS ameliorated CoCl2-induced autophagic flux inhibition in HT22 cells. (A) Levels of LC3, p62, and p-p62 (Ser349) in HT22 cells treated with CQ were assessed, with or without GAS (200 μM) for 24 h (n = 3). (B) Levels of LC3, p62, and phosphorylated p62 (Ser349) in HT22 cells treated with 3-BDO were assessed with or without GAS (200 μM) for 24 h (n = 3). (C) Detection of p62 siRNA transfection efficiency by western blotting. (DF) Levels of LAMP-2, p62, and phosphorylated p62 (Ser349) in HT22 cells transfected with nonspecific or p62 siRNA were evaluated with or without GAS (200 μM) for 24 h (n = 3). The experimental results were normalized to β-actin levels and are shown as fold changes relative to control cells. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments. ##P< 0.01 versus control, *P< 0.05 versus CoCl2. GAS, gastrodin; CoCl2, cobalt chloride; LC3, microtubule-associated protein 1 light chain 3; CQ, chloroquine.