金思维提取物对APPV717I转基因小鼠海马神经元PP2A表达的影响
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摘要
目的：研究金思维提取物(GEPT)对APPV717I转基因小鼠海马神经元蛋白磷酸酶2A(PP2A)表达的影响。
方法：将3月龄的APPV717I转基因小鼠60只随机分为模型组、多奈哌齐组(0.92mg/kg/d)、GEPT小剂量组(0.075g/kg/d)、GEPT中剂量组(0.15g/kg/d)和GEPT大剂量组(0.3g/kg/d),并以同月龄遗传背景相同的C57BL/6J小鼠12只作为正常对照组,治疗组每天灌胃给药1次,正常组及模型组给予0.5%羧甲基纤维素钠溶液灌胃。给药8个月后(11月龄)用免疫组化法和免疫蛋白质印迹法测定海马神经元PP2A的表达水平
结果：
免疫组化染色结果显示模型组小鼠海马神经元CA1区PP2A染色较淡,阳性细胞数和阳性细胞总面积较正常组明显减少(P<0.01)；GEPT小、中、大剂量组小鼠海马神经元CA1区PP2A阳性细胞数和阳性细胞总面积与模型组比较明显增加(P<0.01):GEPT小剂量组小鼠海马神经元CAl区PP2A阳性细胞数和阳性细胞总面积与正常组比较有显著性差异(P<0.05,P<0.01)；GEPT中、大剂量组小鼠海马神经元CA1区PP2A阳性细胞数和阳性细胞总面积与正常组比较无显著性差异(P>0.05),疗效接近多奈哌齐组。
免疫蛋白质印迹结果显示模型组小鼠海马神经元PP2A表达与正常组比较明显减少(P<0.01)；GEPT小、中、大剂量组小鼠海马神经元PP2A表达与模型组比较明显增加(P<0.01)；GEPT小剂量组小鼠海马神经元PP2A表达与正常组比较有显著性差异(P<0.05)；GEPT中、大剂量组小鼠海马神经元PP2A表达与正常组比较无显著性差异(P>0.05),疗效接近多奈哌齐组。该结果和免疫组化结果相一致。
结论：GEPT可增加APPV717I转基因小鼠海马神经元PP2A的表达,提示GEPT可能通过此途径减轻APPV717I转基因小鼠海马神经元tau蛋白的异常过度磷酸化。
Objective:To investigate the effect of GEPT extract on expression of protein phosphatase 2A (PP2A) in hippocampal neurons of APPV717I transgenic mice.
Methods:60 three-month-old APPV717I transgenic mice were randomly divided into model group, donepezil group (0.92mg/kg/d), GEPT low-dose group (0.075g/kg/d), GEPT medium-dose group(0.075g/kg/d) and GEPT high-dose group (0.075g/kg/d), and 12 three-month-old C57BL/6J mice of the same genetic background were set as normal control group, the treatment groups were administered oral ly 1 time a day, and both the control group and model group were given 0.5% sodium carboxymethylcellulose (CMC) solution. Immunohistochemistry and Western blot analysis were used to detect the expression of protein phosphatase 2A (PP2A) in hippocampal neurons of APPV717I transgenic mice after 8 months drug administration (11 months old).
Results:
Immunohistochemical staining of PP2A in hippocampal CA1 region neurons of the model group were thin, and the number of positive cells and the total area of positive cells were significantly decreased compared with that of the control group (P<0.01); the number of pos i t ive cells and the total area of positive cells of the GEPT low, medium and high-dose group were significantly increased compared with that of the model group (P<0.01); the number of positive cells and the total area of positive cells of the GEPT low-dose group showed significant difference compared with that of the control group (P<0.05, P<0.01); the number of positive cells and the total area of positive cells of the GEPT medium and high-dose group showed no significant difference compared with that of the control group (P>0.05), and the effectiveness were similar compared with that of the donepezil group.
Western blot analysis of PP2A expression in hippocampal neurons of the model group were significantly decreased compared with that of the control group (P<0.01); the PP2A expression of the GEPT low, medium and high-dose group were significantly increased compared with that of the model group (P<0.01); the PP2A expression of the GEPT low-dose group showed significant difference compared with that of the control group (P<0.05); the PP2A expression of the GEPT medium and high-dose group showed no significant difference compared with that of the control group (P>0.05), and the effectiveness were similar compared with that of the donepezil group. The results of Western blot analysis were similar to that of Immunohistochemistry.
Conclusion:GEPT can activate the expression of PP2A in hippocampal neurons of APPV717I transgenic mice, which prompt GEPT may attenuate abnormal hyperphosphorylation of tau protein in hippocampal neurons of APPV717I transgenic mice through this way.
方法：将3月龄的APPV717I转基因小鼠60只随机分为模型组、多奈哌齐组(0.92mg/kg/d)、GEPT小剂量组(0.075g/kg/d)、GEPT中剂量组(0.15g/kg/d)和GEPT大剂量组(0.3g/kg/d),并以同月龄遗传背景相同的C57BL/6J小鼠12只作为正常对照组,治疗组每天灌胃给药1次,正常组及模型组给予0.5%羧甲基纤维素钠溶液灌胃。给药8个月后(11月龄)用免疫组化法和免疫蛋白质印迹法测定海马神经元PP2A的表达水平
结果：
免疫组化染色结果显示模型组小鼠海马神经元CA1区PP2A染色较淡,阳性细胞数和阳性细胞总面积较正常组明显减少(P<0.01)；GEPT小、中、大剂量组小鼠海马神经元CA1区PP2A阳性细胞数和阳性细胞总面积与模型组比较明显增加(P<0.01):GEPT小剂量组小鼠海马神经元CAl区PP2A阳性细胞数和阳性细胞总面积与正常组比较有显著性差异(P<0.05,P<0.01)；GEPT中、大剂量组小鼠海马神经元CA1区PP2A阳性细胞数和阳性细胞总面积与正常组比较无显著性差异(P>0.05),疗效接近多奈哌齐组。
免疫蛋白质印迹结果显示模型组小鼠海马神经元PP2A表达与正常组比较明显减少(P<0.01)；GEPT小、中、大剂量组小鼠海马神经元PP2A表达与模型组比较明显增加(P<0.01)；GEPT小剂量组小鼠海马神经元PP2A表达与正常组比较有显著性差异(P<0.05)；GEPT中、大剂量组小鼠海马神经元PP2A表达与正常组比较无显著性差异(P>0.05),疗效接近多奈哌齐组。该结果和免疫组化结果相一致。
结论：GEPT可增加APPV717I转基因小鼠海马神经元PP2A的表达,提示GEPT可能通过此途径减轻APPV717I转基因小鼠海马神经元tau蛋白的异常过度磷酸化。
Objective:To investigate the effect of GEPT extract on expression of protein phosphatase 2A (PP2A) in hippocampal neurons of APPV717I transgenic mice.
Methods:60 three-month-old APPV717I transgenic mice were randomly divided into model group, donepezil group (0.92mg/kg/d), GEPT low-dose group (0.075g/kg/d), GEPT medium-dose group(0.075g/kg/d) and GEPT high-dose group (0.075g/kg/d), and 12 three-month-old C57BL/6J mice of the same genetic background were set as normal control group, the treatment groups were administered oral ly 1 time a day, and both the control group and model group were given 0.5% sodium carboxymethylcellulose (CMC) solution. Immunohistochemistry and Western blot analysis were used to detect the expression of protein phosphatase 2A (PP2A) in hippocampal neurons of APPV717I transgenic mice after 8 months drug administration (11 months old).
Results:
Immunohistochemical staining of PP2A in hippocampal CA1 region neurons of the model group were thin, and the number of positive cells and the total area of positive cells were significantly decreased compared with that of the control group (P<0.01); the number of pos i t ive cells and the total area of positive cells of the GEPT low, medium and high-dose group were significantly increased compared with that of the model group (P<0.01); the number of positive cells and the total area of positive cells of the GEPT low-dose group showed significant difference compared with that of the control group (P<0.05, P<0.01); the number of positive cells and the total area of positive cells of the GEPT medium and high-dose group showed no significant difference compared with that of the control group (P>0.05), and the effectiveness were similar compared with that of the donepezil group.
Western blot analysis of PP2A expression in hippocampal neurons of the model group were significantly decreased compared with that of the control group (P<0.01); the PP2A expression of the GEPT low, medium and high-dose group were significantly increased compared with that of the model group (P<0.01); the PP2A expression of the GEPT low-dose group showed significant difference compared with that of the control group (P<0.05); the PP2A expression of the GEPT medium and high-dose group showed no significant difference compared with that of the control group (P>0.05), and the effectiveness were similar compared with that of the donepezil group. The results of Western blot analysis were similar to that of Immunohistochemistry.
Conclusion:GEPT can activate the expression of PP2A in hippocampal neurons of APPV717I transgenic mice, which prompt GEPT may attenuate abnormal hyperphosphorylation of tau protein in hippocampal neurons of APPV717I transgenic mice through this way.
引文
[1]Ferri CP, Prince M, Brayne C, et al. Global prevalence of dementia: a Delphi consensus study [J]. Lancet,2005,336:2112-2117
[2]van der Flier WM, Scheltens P. Epidemiology and risk factors of dementia [J]. J Neurol Neurosurg Psychiatry,2005,76 Suppl 5:v2-v7
[3]Hardy J, Selkoe DJ. The amyloid hypothesis of Alzheimer's disease: progress and problems on the road to therapeut ics [J]. Science,2002, 297:353-356
[4]Kowalska A. The beta-amyloid cascade hypothesis:a sequence of events leading to neurodegeneration in Alzheimer's disease [J]. Neurol Neurochir Pol,2004,38:405-411
[5]Wirths O, Multhaup G, Bayer TA. A modified beta-amyloid hypothesis: intraneuronal accumulation of the beta-amyloid peptide—the first step of a fatal cascade [J]. J Neurochem,2004,91:513-520
[6]杨济民.老年性痴呆的中医论治[J].中医药研究,2001,17(2)：58-59
[7]王永炎.老年性痴呆辨治[J].中国医药学报,1994,9(2)：49-51
[8]朱未名,胡海燕.阿尔茨海默病的中医药临床研究概况[J].中医杂志,2005,46(5)：389-391
[9]Miao YC, Tian JZ, Shi J, et al. Correlation between cognitive functions and syndromes of traditional Chinese medicine in amnestic mild cognitive impairment [J]. J Chin Integr Med,2009,7(3):205-211
[10]田金洲,时晶,王永炎,等.阿尔茨海默病的诊断与治疗[M].北京：人民卫生出版社,2009：213-215
[11]张沁园,魏久贞.升黄益智颗粒治疗阿尔茨海默病临床观察[J].山东中医药大学学报,2008,32(4)：303-305
[12]董克礼,曾望远.益智健脑颗粒治疗阿尔茨海默氏病20例总结[J].湖南中医杂志,2006,22(2)：3-4
[13]李亚明.中医药防治阿尔茨海默病的思路与方法[J].中国临床康复,2005,9(24)：144-146
[14]Allain H, Ferrer DB, Tribut O, et al. Alzheimer's disease:the pharmacological pathway. Fundam [J]. Clin Pharmacol,2003,17: 419-428
[15]Tian J, Xu Y, Sheng SL, et al. Effect of GETO extract on expression of ChAT and NGF in the brain with AD model [J]. Alzheimer's and Dementia,2006,2:s601-s605
[16]Tohda C, Tamura T, Komatsu K. Repair of amyloid β 25-35 induced memory impairment and synaptic loss by a Kampo formula, Zokumeito [J]. Brain Res,2003,990:141-147
[17]Egashir N, Iwasaki K, Akiyoshi Y, et al. Protective effect of Tokishakuyaku-san on amyloid β 25-35 induced neuronal damage in cultured rat cortical neurons [J]. Phytother Res,2005,19:450-45
[18]Koo BS, Kim YK, Park KS. Attenuating effect of a traditional Korean formulation, Paeng-Jo-Yeon-Nyeon-Baek-Ja-In-Hwan (PJBH), on hydrogen peroxide induced injury in PC12 cells [J]. Phytother Res, 2004,18:488-493
[19]Misiti F, Sampaolese B, Mezzogori D, et al. Protective effect of rhubarb derivatives on amyloid betal-42 peptide-induced apoptosis in IMR-32 cells:A case of nutrigenomic [J]. Brain Res Bull,2006, 71:29-36
[20]Chauhan NB. Effect of aged garlic extract on APP processing and tau phosphorylation in Alzheimer's transgenic model Tg2576 [J]. J Ethnopharmacol,2006,108:385-394
[21]Tohda C, Matsumoto N, Zou K, et al. A beta25-35 induced memory impairment, axonal atrophy, and synaptic loss are ameliorated by M1, Ametabolite of protopanaxadiol type saponins [J]. Neuropsycho-pharmacol,2004,29:860-868
[1]Riley KP, Snowdon DA, Markesbery WR. Alzheimer's neurofibrillary pathology and the spectrum of cognitive function:findings from the Nun Study [J]. Ann Neurol,2002,51(5):567-577
[2]Alonso A, Zaidi T, Novak M, et al. Hyperphosphorylation induces self-assembly of tau into tangles of paired helical filaments/ straight filaments [J]. Proc Nati Acad Sci USA,2001,98(12): 6923-6928
[3]Perez M, Hemandez F, Gomez-Ramos A, et al. Formation of aberrant phospho tau fibrillar polymers in neural cultured cells [J]. Eur J Biochem,2002,269(5):1484-1489
[4]Robert M, Mathuranath PS. Tau and tauopathies [J]. Neurol India,2007, 55(1):11-16
[5]Trojanowski JQ, Lee VM. The role of tau in Alzheimer's disease [J]. Med Clin North Am,2002,86:615-627
[6]Garcia ML, Cleveland DW. Going new places using an old MAP:tau, microtubules and human neurodegenerat ive di sease [J]. Curr Opin Cell Biol,2001,13(1):41-48
[7]Johnson GV, Stoothoff WH. Tau phosphorylation in neuronal cell function and dysfunction [J]. J Cell Sci,2004,117:5721-5729
[8]Alonso AD, Clerico JD, Li B, et al. Phosphorylation of Tau at Thr212, Thr231, and Ser262 combined causes neurodegeneration [J]. J Bio Chem, 2010,285:30851-30860
[9]Sun L, Wang X, Liu S, et al. Bilateral injection of isoproterenol into hippocampus induces Alzheimer-like hyperphosphorylation of tau and spatial memory deficit in rat [J]. FEBS Lett,2005,579(1): 251-258
[10]Liu SJ, Zhang JY, Li HL, et al. Tau becomes a more favorable substrate for GSK-3 when it is prephosphorylated by PKA in rat brain [J]. J Biol Chem,2004,279(48):50078-50088
[11]Gong CX, Liu F, Grundke-Iqbal I, et al. Post-translational modify-cations of tau protein in Alzheimer's disease [J]. Neural Transm, 2005,112(6):813-838
[12]Liu F, Grundke-Iqbal I, Iqbal K, et al. Contributions of protein phosphatases PP1, PP2A, PP2B and PP5 to the regulation of tau phosphorylation [J]. Eur J Neurosci,2005,22(8):1942-1950
[13]Liu F, Iqbal K, Grundke-Iqbal I, et al. Dephosphorylation of tau by protein phosphatase 5:Impairment in Alzheimer's disease [J]. J Biol Chem,2005,280(3):1790-1796
[14]Pei JJ, Gong CX, An WL, et al. Okadaic-acid-induced inhibition of protein phosphatase 2A produces activation of mitogen-activated protein kinases ERK1/2, MEK1/2, and P70 S6, similar to that in Alzheimer's disease [J]. Am J Pathol,2003,163(3):845-858
[15]Tanimukai H, Grundke-Iqbal I, Iqbal K. Up-regulation of inhibitors of pprotein phosphatase 2A in Alzheimer's disease [J]. Am J Pathol, 2005,166(6):1761-1771
[16]Avila J, Lucas JJ, Perez M, et al. Role of tau protein in both physiological and pathological conditions [J]. Physiol Rev,2004, 84(2),361-384
[17]Wang XC, Wu YG, Hu ZH, et al. Correlation of Alzheimer-like tau hyperphosphorylation and fMRI bold intensity [J]. J Current Alzheimer Res,2004,1(2):143-145
[18]Goldbaum 0, Oppermann M, Handschu HM, et al. Proteasome inhibition stabilizes tau inclusions in oligodendroglial cells that occur after treatment with okadaic acid [J]. Neurosci,2003,23(26): 8872-8880
[19]Keck S, Nitsch R, Grune T, et al. Proteasome inhibition by paired helical filament-tau in brains of patients with Alzheimer's Disease [J]. J Neurochem,2003,85(1):115-122
[20]Ballatore C, Lee VMY, Trojanowski JQ. Tau-mediated neurodegene-ration in Alzheimer's disease and related disorders [J]. Nat Rev Neurosci,2007,8:663-672
[21]Haass C, Mandelkow E. Fyn-Tau-Amyloid:A toxic triad [J]. Cell,2010, 142(3):356-358
[22]Gomez-Ramos A, Diaz-Hernandez M, Rubio A, et al. Extracellular tau promotes intracellular calcium increase through M1 and M3 musca-rinic receptors in neuronal cells [J]. Mol Cell Neurosci,2008, 37(4):673-681
[23]Craig-Schapiro R, Fagan AM, Holtzman DM. Biomarkers of Alzheimer' s disease [J]. Neurobiol Dis,2009,35(2):128-140
[24]Hampel H, Buerger K, Zinkowski R, et al. Measurement of phosphory-lated tau epitopes in the differential diagnosis of Alzheimer disease:a comparative cerebrospinal fluid study [J]. Arch Gen Psychiatry,2004,61(1):95-102
[25]Gong CX, Iqbal K. Hyperphosphorylation of microtubule-associated protein tau:a promising therapeutic target for Alzheimer disease [J]. Curr Med Chem,2008,15(23):2321-2328
[26]Avila J. Tau phosphorylation and aggregation in Alzheimer's disease pathology [J]. FEBS Lett,2006,580(12):2922-2927
[27]Iqbal K, Grundke-Iqbal I. Developing pharmacological therapies for Alzheimer disease [J]. Cell Mol Life Sci,2007,64(17):2234-2244
[28]Churcher I. Tau therapeutic strategies for the treatment of Alzheimer's disease [J]. Curr Top Med Chem,2006,6(6):579-595
[29]Golde TE. Disease modifying therapy for AD? [J]. J Neurochem,2006, 99(3):689-707
[30]Roder HM, Hut ton ML. Microtubule-associated protein tau as a therapeutic target in neurodegenerative disease [J]. Expert Opin Ther Targets,2007,11(4):435-442
[31]Brunden KR, Trojanowski JQ, Lee VMY. Advances in tau-focused drug discovery for Alzheimer's disease and related tauopathies [J]. Nat Rev Drug Discov,2009,8(10):783-793
[32]Luo J, Yin JH, Wei Q. The effect of calcineurin act ivator, extracted from Chinese herbal medicine, on memory and immunity in mice [J]. Pharmacol Biochem Behav,2003,75(4):749-754
[33]Sigurdsson EM. Immunotherapy target ing pathological tau protein in Alzheimer's disease and related tauopathies [J]. J Alzheimers Dis, 2008,15(2):157-168
[34]Kayed R, Jackson GR. Prefilament tau species as potential targets for immunotherapy for Alzheimer disease and related disorders [J]. Curr Opin Immunol,2009,21(3):359-363
[1]Riley KP, Snowdon DA, Markesbery WR. Alzheimer's neurofibrillary pathology and the spectrum of cognitive function:findings from the Nun Study [J]. Ann Neurol,2002,51(5):567-577
[2]Gong CX, Liu F, Grundke-Iqbal I, et al. Post-translational modify-cations of tau protein in Alzheimer's disease [J]. J Neural Transm, 2005,112(6):813-838
[3]陈玉静,王蓉,盛树力,等.复方金思维对老年性痴呆模型大鼠海马神经元微管和Tau蛋白磷酸化相关酶类表达的影响[J].天津中医药,2008,25(1)：59-62
[4]Perez M, Hemandez F, Gomez-Ramos A, et al. Formation of aberrant phospho tau fibrillar polymers in neural cultured cells [J]. Eur J Biochem,2002,269(5):1484-1489
[5]Gomez-Ramos A, Diaz-Hernandez M, Cuadros R, et al. Extracellular tau is toxic to neuronal cells [J]. FEBS Lett,2006,580(20):4842-4850
[6]Gomez-Ramos A, Diaz-Hernandez M, Rubio A, et al. Extracellular tau promotes iniraoellulai calcium increase through M1 and M3 muscarinic receptors in neuronal cells [J]. Mol Cell Neurosci,2008,37(4): 673-681
[7]Ballatore C, Lee VMY, Trojanowski JQ. Tau-mediated neurodegene-ration in Alzheimer's disease and related disorders [J]. Nat Rev Neurosci,2007,8(9):663-672
[8]Toole BJ, Cohen PT. The skeletal muscle-specific glycogen-targeted protein phosphatase 1 plays a major role in the regulation of glycogen metabolism by adrenaline in vivo [J]. Cell Signal,2007, 19(5):1044-1055
[9]Lauwaet T, Davids BJ, Torres-Escobar A, et al. Protein phosphatase 2A play a crucial role in Giardia differentiation [J]. Mol Biochem Parasitolo,2007,152(1):80-89
[10]Klee CB, Ren H, Wang X. Regulation of the calmodulin-stimulated protein phosphatase, calcineurin [J]. J Biol Chem,1998,273(22): 13367-13370
[11]Groves MR, Hanlon N, Turowski P, et al. The structure of the protein phosphatase 2A PR65/A subunit reveals the conformation of its 15 tandemly repeated HEAT motifs [J]. Cell,1999,96(1):99-110
[12]Maynes JT, Luu HA, Cherney MM, et al. Crystal structure of protein phosphatase-1 bound to Motuporin and Dihydromicro-cystin-LA: Elucidation of the mechanism of enzyme inhibition by cyanobacterial toxins [J]. J Mol Biol,2006,356(10):111-120
[13]Xu YH, Xing YN, Chen Y, et al. Structure of the protein phosphatase2A holoenzyme [J]. Cell,2006,127(15):1239-1251
[14]Xing YN, Xu YH, Chen Y, et al. Structure of protein phosphatase2A core enzyme bound to tumor-Inducing toxins [J]. Cell,2006,127(20): 341-353
[15]Wu Y, Song P, Xu J, et al. Activation of protein phosphatase 2A by palmitate inhibits AMP-activated protein kinase [J]. J Biol Chem, 2007,282(13):9777-9788
[16]Liu F, Grundke-Iqbal I, Iqbal K, et al. Contributions of protein phosphatases PP1, PP2A, PP2B and PP5 to the regulation of tau phosphorylation [J]. Eur J Neurosci,2005,22(8):1942-1950
[17]Walton JR. An aluminum-based rat model for Alzheimer's disease exhibits oxidative damage, inhibition of PP2A activity, hyperphos-phorylated tau and granulovacular degeneration [J]. J Inorg Biochem, 2007,101(9):1275-1284
[18]Pei JJ, Gong CX, An WL, et al. Okadaic-acid-induced inhibition of protein phosphatase 2A produces activation of mi togen-activated protein kinases ERK1/2, MEK1/2, and P70 S6, similar to that in Alzheimer's disease [J]. Am J Pathol,2003,163(3):845-858
[19]Gong CX, Shaikh S, Wang JZ, et al. Phosphatase activity towards abnormally phosphorylated tau:decrease in Alzheimer disease brain [J]. J Neurochem,1995,65:732-738
[20]Pei JJ, Serson E, lqbal K, et al. Expression of protein phosphatases (PP1. PP2A. PP2B and PTP1B) and protein kinases (MAP kinase and P43-cdc2) in the hippocampus of patients with Alzheimer disease and normal aged individuals [J]. Brain Res,1994,655:70-76
[21]Tanaka T, lqbal K, Trenkner E, et al. Abnormally phosphorylated tau in SY5Y human neuroblastoma cells [J]. FEBS lett,1995,360:5-9
[22]Miao YC, Tian JZ, Shi J, et al. Correlation between cognitive functions and syndromes of traditional Chinese medicine in amnestic mild cognitive impairment [J]. J Chin Integr Med,2009,7(3): 205-211
[23]Tian J, Shi J, Zhang L, et al. GEPT extract reduces Abeta deposition by regulating the balance between production and degradation of Abeta in APPV717I transgenic mice [J]. Curr Alzheimer Res,2009, 6(2):118-131
[2]van der Flier WM, Scheltens P. Epidemiology and risk factors of dementia [J]. J Neurol Neurosurg Psychiatry,2005,76 Suppl 5:v2-v7
[3]Hardy J, Selkoe DJ. The amyloid hypothesis of Alzheimer's disease: progress and problems on the road to therapeut ics [J]. Science,2002, 297:353-356
[4]Kowalska A. The beta-amyloid cascade hypothesis:a sequence of events leading to neurodegeneration in Alzheimer's disease [J]. Neurol Neurochir Pol,2004,38:405-411
[5]Wirths O, Multhaup G, Bayer TA. A modified beta-amyloid hypothesis: intraneuronal accumulation of the beta-amyloid peptide—the first step of a fatal cascade [J]. J Neurochem,2004,91:513-520
[6]杨济民.老年性痴呆的中医论治[J].中医药研究,2001,17(2)：58-59
[7]王永炎.老年性痴呆辨治[J].中国医药学报,1994,9(2)：49-51
[8]朱未名,胡海燕.阿尔茨海默病的中医药临床研究概况[J].中医杂志,2005,46(5)：389-391
[9]Miao YC, Tian JZ, Shi J, et al. Correlation between cognitive functions and syndromes of traditional Chinese medicine in amnestic mild cognitive impairment [J]. J Chin Integr Med,2009,7(3):205-211
[10]田金洲,时晶,王永炎,等.阿尔茨海默病的诊断与治疗[M].北京：人民卫生出版社,2009：213-215
[11]张沁园,魏久贞.升黄益智颗粒治疗阿尔茨海默病临床观察[J].山东中医药大学学报,2008,32(4)：303-305
[12]董克礼,曾望远.益智健脑颗粒治疗阿尔茨海默氏病20例总结[J].湖南中医杂志,2006,22(2)：3-4
[13]李亚明.中医药防治阿尔茨海默病的思路与方法[J].中国临床康复,2005,9(24)：144-146
[14]Allain H, Ferrer DB, Tribut O, et al. Alzheimer's disease:the pharmacological pathway. Fundam [J]. Clin Pharmacol,2003,17: 419-428
[15]Tian J, Xu Y, Sheng SL, et al. Effect of GETO extract on expression of ChAT and NGF in the brain with AD model [J]. Alzheimer's and Dementia,2006,2:s601-s605
[16]Tohda C, Tamura T, Komatsu K. Repair of amyloid β 25-35 induced memory impairment and synaptic loss by a Kampo formula, Zokumeito [J]. Brain Res,2003,990:141-147
[17]Egashir N, Iwasaki K, Akiyoshi Y, et al. Protective effect of Tokishakuyaku-san on amyloid β 25-35 induced neuronal damage in cultured rat cortical neurons [J]. Phytother Res,2005,19:450-45
[18]Koo BS, Kim YK, Park KS. Attenuating effect of a traditional Korean formulation, Paeng-Jo-Yeon-Nyeon-Baek-Ja-In-Hwan (PJBH), on hydrogen peroxide induced injury in PC12 cells [J]. Phytother Res, 2004,18:488-493
[19]Misiti F, Sampaolese B, Mezzogori D, et al. Protective effect of rhubarb derivatives on amyloid betal-42 peptide-induced apoptosis in IMR-32 cells:A case of nutrigenomic [J]. Brain Res Bull,2006, 71:29-36
[20]Chauhan NB. Effect of aged garlic extract on APP processing and tau phosphorylation in Alzheimer's transgenic model Tg2576 [J]. J Ethnopharmacol,2006,108:385-394
[21]Tohda C, Matsumoto N, Zou K, et al. A beta25-35 induced memory impairment, axonal atrophy, and synaptic loss are ameliorated by M1, Ametabolite of protopanaxadiol type saponins [J]. Neuropsycho-pharmacol,2004,29:860-868
[1]Riley KP, Snowdon DA, Markesbery WR. Alzheimer's neurofibrillary pathology and the spectrum of cognitive function:findings from the Nun Study [J]. Ann Neurol,2002,51(5):567-577
[2]Alonso A, Zaidi T, Novak M, et al. Hyperphosphorylation induces self-assembly of tau into tangles of paired helical filaments/ straight filaments [J]. Proc Nati Acad Sci USA,2001,98(12): 6923-6928
[3]Perez M, Hemandez F, Gomez-Ramos A, et al. Formation of aberrant phospho tau fibrillar polymers in neural cultured cells [J]. Eur J Biochem,2002,269(5):1484-1489
[4]Robert M, Mathuranath PS. Tau and tauopathies [J]. Neurol India,2007, 55(1):11-16
[5]Trojanowski JQ, Lee VM. The role of tau in Alzheimer's disease [J]. Med Clin North Am,2002,86:615-627
[6]Garcia ML, Cleveland DW. Going new places using an old MAP:tau, microtubules and human neurodegenerat ive di sease [J]. Curr Opin Cell Biol,2001,13(1):41-48
[7]Johnson GV, Stoothoff WH. Tau phosphorylation in neuronal cell function and dysfunction [J]. J Cell Sci,2004,117:5721-5729
[8]Alonso AD, Clerico JD, Li B, et al. Phosphorylation of Tau at Thr212, Thr231, and Ser262 combined causes neurodegeneration [J]. J Bio Chem, 2010,285:30851-30860
[9]Sun L, Wang X, Liu S, et al. Bilateral injection of isoproterenol into hippocampus induces Alzheimer-like hyperphosphorylation of tau and spatial memory deficit in rat [J]. FEBS Lett,2005,579(1): 251-258
[10]Liu SJ, Zhang JY, Li HL, et al. Tau becomes a more favorable substrate for GSK-3 when it is prephosphorylated by PKA in rat brain [J]. J Biol Chem,2004,279(48):50078-50088
[11]Gong CX, Liu F, Grundke-Iqbal I, et al. Post-translational modify-cations of tau protein in Alzheimer's disease [J]. Neural Transm, 2005,112(6):813-838
[12]Liu F, Grundke-Iqbal I, Iqbal K, et al. Contributions of protein phosphatases PP1, PP2A, PP2B and PP5 to the regulation of tau phosphorylation [J]. Eur J Neurosci,2005,22(8):1942-1950
[13]Liu F, Iqbal K, Grundke-Iqbal I, et al. Dephosphorylation of tau by protein phosphatase 5:Impairment in Alzheimer's disease [J]. J Biol Chem,2005,280(3):1790-1796
[14]Pei JJ, Gong CX, An WL, et al. Okadaic-acid-induced inhibition of protein phosphatase 2A produces activation of mitogen-activated protein kinases ERK1/2, MEK1/2, and P70 S6, similar to that in Alzheimer's disease [J]. Am J Pathol,2003,163(3):845-858
[15]Tanimukai H, Grundke-Iqbal I, Iqbal K. Up-regulation of inhibitors of pprotein phosphatase 2A in Alzheimer's disease [J]. Am J Pathol, 2005,166(6):1761-1771
[16]Avila J, Lucas JJ, Perez M, et al. Role of tau protein in both physiological and pathological conditions [J]. Physiol Rev,2004, 84(2),361-384
[17]Wang XC, Wu YG, Hu ZH, et al. Correlation of Alzheimer-like tau hyperphosphorylation and fMRI bold intensity [J]. J Current Alzheimer Res,2004,1(2):143-145
[18]Goldbaum 0, Oppermann M, Handschu HM, et al. Proteasome inhibition stabilizes tau inclusions in oligodendroglial cells that occur after treatment with okadaic acid [J]. Neurosci,2003,23(26): 8872-8880
[19]Keck S, Nitsch R, Grune T, et al. Proteasome inhibition by paired helical filament-tau in brains of patients with Alzheimer's Disease [J]. J Neurochem,2003,85(1):115-122
[20]Ballatore C, Lee VMY, Trojanowski JQ. Tau-mediated neurodegene-ration in Alzheimer's disease and related disorders [J]. Nat Rev Neurosci,2007,8:663-672
[21]Haass C, Mandelkow E. Fyn-Tau-Amyloid:A toxic triad [J]. Cell,2010, 142(3):356-358
[22]Gomez-Ramos A, Diaz-Hernandez M, Rubio A, et al. Extracellular tau promotes intracellular calcium increase through M1 and M3 musca-rinic receptors in neuronal cells [J]. Mol Cell Neurosci,2008, 37(4):673-681
[23]Craig-Schapiro R, Fagan AM, Holtzman DM. Biomarkers of Alzheimer' s disease [J]. Neurobiol Dis,2009,35(2):128-140
[24]Hampel H, Buerger K, Zinkowski R, et al. Measurement of phosphory-lated tau epitopes in the differential diagnosis of Alzheimer disease:a comparative cerebrospinal fluid study [J]. Arch Gen Psychiatry,2004,61(1):95-102
[25]Gong CX, Iqbal K. Hyperphosphorylation of microtubule-associated protein tau:a promising therapeutic target for Alzheimer disease [J]. Curr Med Chem,2008,15(23):2321-2328
[26]Avila J. Tau phosphorylation and aggregation in Alzheimer's disease pathology [J]. FEBS Lett,2006,580(12):2922-2927
[27]Iqbal K, Grundke-Iqbal I. Developing pharmacological therapies for Alzheimer disease [J]. Cell Mol Life Sci,2007,64(17):2234-2244
[28]Churcher I. Tau therapeutic strategies for the treatment of Alzheimer's disease [J]. Curr Top Med Chem,2006,6(6):579-595
[29]Golde TE. Disease modifying therapy for AD? [J]. J Neurochem,2006, 99(3):689-707
[30]Roder HM, Hut ton ML. Microtubule-associated protein tau as a therapeutic target in neurodegenerative disease [J]. Expert Opin Ther Targets,2007,11(4):435-442
[31]Brunden KR, Trojanowski JQ, Lee VMY. Advances in tau-focused drug discovery for Alzheimer's disease and related tauopathies [J]. Nat Rev Drug Discov,2009,8(10):783-793
[32]Luo J, Yin JH, Wei Q. The effect of calcineurin act ivator, extracted from Chinese herbal medicine, on memory and immunity in mice [J]. Pharmacol Biochem Behav,2003,75(4):749-754
[33]Sigurdsson EM. Immunotherapy target ing pathological tau protein in Alzheimer's disease and related tauopathies [J]. J Alzheimers Dis, 2008,15(2):157-168
[34]Kayed R, Jackson GR. Prefilament tau species as potential targets for immunotherapy for Alzheimer disease and related disorders [J]. Curr Opin Immunol,2009,21(3):359-363
[1]Riley KP, Snowdon DA, Markesbery WR. Alzheimer's neurofibrillary pathology and the spectrum of cognitive function:findings from the Nun Study [J]. Ann Neurol,2002,51(5):567-577
[2]Gong CX, Liu F, Grundke-Iqbal I, et al. Post-translational modify-cations of tau protein in Alzheimer's disease [J]. J Neural Transm, 2005,112(6):813-838
[3]陈玉静,王蓉,盛树力,等.复方金思维对老年性痴呆模型大鼠海马神经元微管和Tau蛋白磷酸化相关酶类表达的影响[J].天津中医药,2008,25(1)：59-62
[4]Perez M, Hemandez F, Gomez-Ramos A, et al. Formation of aberrant phospho tau fibrillar polymers in neural cultured cells [J]. Eur J Biochem,2002,269(5):1484-1489
[5]Gomez-Ramos A, Diaz-Hernandez M, Cuadros R, et al. Extracellular tau is toxic to neuronal cells [J]. FEBS Lett,2006,580(20):4842-4850
[6]Gomez-Ramos A, Diaz-Hernandez M, Rubio A, et al. Extracellular tau promotes iniraoellulai calcium increase through M1 and M3 muscarinic receptors in neuronal cells [J]. Mol Cell Neurosci,2008,37(4): 673-681
[7]Ballatore C, Lee VMY, Trojanowski JQ. Tau-mediated neurodegene-ration in Alzheimer's disease and related disorders [J]. Nat Rev Neurosci,2007,8(9):663-672
[8]Toole BJ, Cohen PT. The skeletal muscle-specific glycogen-targeted protein phosphatase 1 plays a major role in the regulation of glycogen metabolism by adrenaline in vivo [J]. Cell Signal,2007, 19(5):1044-1055
[9]Lauwaet T, Davids BJ, Torres-Escobar A, et al. Protein phosphatase 2A play a crucial role in Giardia differentiation [J]. Mol Biochem Parasitolo,2007,152(1):80-89
[10]Klee CB, Ren H, Wang X. Regulation of the calmodulin-stimulated protein phosphatase, calcineurin [J]. J Biol Chem,1998,273(22): 13367-13370
[11]Groves MR, Hanlon N, Turowski P, et al. The structure of the protein phosphatase 2A PR65/A subunit reveals the conformation of its 15 tandemly repeated HEAT motifs [J]. Cell,1999,96(1):99-110
[12]Maynes JT, Luu HA, Cherney MM, et al. Crystal structure of protein phosphatase-1 bound to Motuporin and Dihydromicro-cystin-LA: Elucidation of the mechanism of enzyme inhibition by cyanobacterial toxins [J]. J Mol Biol,2006,356(10):111-120
[13]Xu YH, Xing YN, Chen Y, et al. Structure of the protein phosphatase2A holoenzyme [J]. Cell,2006,127(15):1239-1251
[14]Xing YN, Xu YH, Chen Y, et al. Structure of protein phosphatase2A core enzyme bound to tumor-Inducing toxins [J]. Cell,2006,127(20): 341-353
[15]Wu Y, Song P, Xu J, et al. Activation of protein phosphatase 2A by palmitate inhibits AMP-activated protein kinase [J]. J Biol Chem, 2007,282(13):9777-9788
[16]Liu F, Grundke-Iqbal I, Iqbal K, et al. Contributions of protein phosphatases PP1, PP2A, PP2B and PP5 to the regulation of tau phosphorylation [J]. Eur J Neurosci,2005,22(8):1942-1950
[17]Walton JR. An aluminum-based rat model for Alzheimer's disease exhibits oxidative damage, inhibition of PP2A activity, hyperphos-phorylated tau and granulovacular degeneration [J]. J Inorg Biochem, 2007,101(9):1275-1284
[18]Pei JJ, Gong CX, An WL, et al. Okadaic-acid-induced inhibition of protein phosphatase 2A produces activation of mi togen-activated protein kinases ERK1/2, MEK1/2, and P70 S6, similar to that in Alzheimer's disease [J]. Am J Pathol,2003,163(3):845-858
[19]Gong CX, Shaikh S, Wang JZ, et al. Phosphatase activity towards abnormally phosphorylated tau:decrease in Alzheimer disease brain [J]. J Neurochem,1995,65:732-738
[20]Pei JJ, Serson E, lqbal K, et al. Expression of protein phosphatases (PP1. PP2A. PP2B and PTP1B) and protein kinases (MAP kinase and P43-cdc2) in the hippocampus of patients with Alzheimer disease and normal aged individuals [J]. Brain Res,1994,655:70-76
[21]Tanaka T, lqbal K, Trenkner E, et al. Abnormally phosphorylated tau in SY5Y human neuroblastoma cells [J]. FEBS lett,1995,360:5-9
[22]Miao YC, Tian JZ, Shi J, et al. Correlation between cognitive functions and syndromes of traditional Chinese medicine in amnestic mild cognitive impairment [J]. J Chin Integr Med,2009,7(3): 205-211
[23]Tian J, Shi J, Zhang L, et al. GEPT extract reduces Abeta deposition by regulating the balance between production and degradation of Abeta in APPV717I transgenic mice [J]. Curr Alzheimer Res,2009, 6(2):118-131