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 the CoCl2-induced suppression of autophagic flux by inhibiting [Ca2+]i-dependent CaMKII phosphorylation in HT22 cells. (G) Coimmunostaining of LC3 with LAMP2 in HT22 cells. LC3+ puncta (green puncta) and LC3+LAMP2+ puncta (yellow puncta) were visualized using a Leica DMIRB at 800× magnification. In each independent experiment, 5 visual field cells were randomly selected and quantified. (H–J) The interaction between CaMKII and p62 was analyzed using immunoprecipitation and mass spectrometry. Data are presented as the mean ± SEM from three independent experiments. ##P*P2, $p2 + CaMKII-siRNA. GAS, gastrodin; LC3, microtubule-associated protein 1 light chain 3; LAMP-2, lysosomal-associated membrane protein-2; CaMKII, Ca2+-calmodulin stimulated protein kinase II.

Figure 6. GAS alleviated the CoCl2-induced suppression of autophagic flux by inhibiting [Ca2+]i-dependent CaMKII phosphorylation in HT22 cells. (G) Coimmunostaining of LC3 with LAMP2 in HT22 cells. LC3+ puncta (green puncta) and LC3+LAMP2+ puncta (yellow puncta) were visualized using a Leica DMIRB at 800× magnification. In each independent experiment, 5 visual field cells were randomly selected and quantified. (HJ) The interaction between CaMKII and p62 was analyzed using immunoprecipitation and mass spectrometry. Data are presented as the mean ± SEM from three independent experiments. ##P< 0.01 versus control, *P< 0.05 versus CoCl2, $p<0.05 versus CoCl2 + CaMKII-siRNA. GAS, gastrodin; LC3, microtubule-associated protein 1 light chain 3; LAMP-2, lysosomal-associated membrane protein-2; CaMKII, Ca2+-calmodulin stimulated protein kinase II.