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 alleviated CoCl2-induced intracellular Ca2+ abundance and CaMKII activation. (A) After HT22 cells were treated with CoCl2 for different time points, the Ca2+ content was detected using a commercial calcium quantitative kit. (B) After HT22 cells were treated with different doses of CoCl2 for 24 h, phosphorylated CaMKII α increases in a dose-dependent manner. β-actin was used as a loading control. (C) HT22 cells were pretreated with GAS and BAPT-AM (0.5 μM) for 1 h and then plated with CoCl2 (200 μM) for 24 h; the Ca2+ content was detected using a commercial calcium quantitative kit. (D) HT22 cells were pretreated with GAS for 1 h and exposed to CoCl2 (200 μM) for 24 h; cytosolic Ca2+ levels were measured by flow cytometry. (E) GAS, similar to calcium chelator (BAPTA-AM), can reduce the level of phosphorylated CaMKII. (F–G) Levels of CaMKII and phosphorylated CaMKII (Ser249) in HT22 cells treated with calcium chelator (BAPTA-AM) or calcium ionophore (A23187) were detected with or without GAS treatment (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; CaMKII, Ca2+-calmodulin stimulated protein kinase II.

Figure 5. GAS alleviated CoCl2-induced intracellular Ca2+ abundance and CaMKII activation. (A) After HT22 cells were treated with CoCl2 for different time points, the Ca2+ content was detected using a commercial calcium quantitative kit. (B) After HT22 cells were treated with different doses of CoCl2 for 24 h, phosphorylated CaMKII α increases in a dose-dependent manner. β-actin was used as a loading control. (C) HT22 cells were pretreated with GAS and BAPT-AM (0.5 μM) for 1 h and then plated with CoCl2 (200 μM) for 24 h; the Ca2+ content was detected using a commercial calcium quantitative kit. (D) HT22 cells were pretreated with GAS for 1 h and exposed to CoCl2 (200 μM) for 24 h; cytosolic Ca2+ levels were measured by flow cytometry. (E) GAS, similar to calcium chelator (BAPTA-AM), can reduce the level of phosphorylated CaMKII. (FG) Levels of CaMKII and phosphorylated CaMKII (Ser249) in HT22 cells treated with calcium chelator (BAPTA-AM) or calcium ionophore (A23187) were detected with or without GAS treatment (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; CaMKII, Ca2+-calmodulin stimulated protein kinase II.