OBJECTIVE The aim of the present study was to investigate the effect of cornel iridoid glycoside(CIG)on tau hyperphosphorylation induced by wortmannin(WT)and GF-109203X(GFX)and the underlying mechanisms.METHODS Human ...OBJECTIVE The aim of the present study was to investigate the effect of cornel iridoid glycoside(CIG)on tau hyperphosphorylation induced by wortmannin(WT)and GF-109203X(GFX)and the underlying mechanisms.METHODS Human neuroblastoma SK-N-SH cells were pre-incubated with CIG(50,100,and 200μg·mL-1,respectively)for 24 h,and then exposed to WT 10μmol·L-1 and GFX 10μmol·L-1for 3hafter washing out CIG.Immuno-fluorescence was used to observe the microtubular cytoskeleton of the cultured cells.Western blotting was used to measure the phosphorylation level of tau protein,glycogen synthase kinase 3β(GSK-3β)and protein phosphatase 2A(PP2A).The activity of PP2 Awas detected by a biochemical assay.RESULTS Pre-incubation of CIG significantly attenuated the WT/GFX-induced tau hyperphosphorylation at the sites of Thr205,Thr212,Ser214,Thr217Ser396 and PHF-1,and improved the damage of morphology and microtubular cytoskeleton of the cells.CIG did not prevent the decrease in p-AKT-ser473 and pGSK3β-ser9 induced by WT/GFX.However,CIG significantly elevated the activity of PP2 Aby reducing the demethylation of PP2AC at Leu309 and the ratio of PME-1/LCMT in the WT/GFX-treated cells.CONCLUSION CIG obviously attenuated WT/GFX-induced tau hyper-phosphorylation at multiple AD-related sites by increasing the activity of PP2A.The mechanism may be involved in the reduced demethylation of PP2 Avia down regulating the ratio of PME/LCMT.展开更多
基金The project supported by National Natural Science Foundation of China(81274120,81273498)the Beijing Natural Science Foundation(7132110)+1 种基金the Beijing Science and Technology Program(Z131102002813066)the Capital Health Development Scientific Grant(2011-1001-05)
文摘OBJECTIVE The aim of the present study was to investigate the effect of cornel iridoid glycoside(CIG)on tau hyperphosphorylation induced by wortmannin(WT)and GF-109203X(GFX)and the underlying mechanisms.METHODS Human neuroblastoma SK-N-SH cells were pre-incubated with CIG(50,100,and 200μg·mL-1,respectively)for 24 h,and then exposed to WT 10μmol·L-1 and GFX 10μmol·L-1for 3hafter washing out CIG.Immuno-fluorescence was used to observe the microtubular cytoskeleton of the cultured cells.Western blotting was used to measure the phosphorylation level of tau protein,glycogen synthase kinase 3β(GSK-3β)and protein phosphatase 2A(PP2A).The activity of PP2 Awas detected by a biochemical assay.RESULTS Pre-incubation of CIG significantly attenuated the WT/GFX-induced tau hyperphosphorylation at the sites of Thr205,Thr212,Ser214,Thr217Ser396 and PHF-1,and improved the damage of morphology and microtubular cytoskeleton of the cells.CIG did not prevent the decrease in p-AKT-ser473 and pGSK3β-ser9 induced by WT/GFX.However,CIG significantly elevated the activity of PP2 Aby reducing the demethylation of PP2AC at Leu309 and the ratio of PME-1/LCMT in the WT/GFX-treated cells.CONCLUSION CIG obviously attenuated WT/GFX-induced tau hyper-phosphorylation at multiple AD-related sites by increasing the activity of PP2A.The mechanism may be involved in the reduced demethylation of PP2 Avia down regulating the ratio of PME/LCMT.
