不同中医治法对AD模型大鼠海马区GSK-3β表达的影响
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摘要
目的：
研究补肾活血法(益智健脑颗粒)、补肾法(地黄饮子)和活血法(血府逐瘀汤)治疗对AD模型大鼠学习、记忆能力及海马区糖原合酶激酶-3β(GSK-3β)表达的影响,从而探讨补肾活血法治疗AD的优势及部分作用机制。
方法：
将60只雄性SD大鼠按体重随机分为6组,分别为正常组、假手术组、模型组、补肾活血组、补肾组、活血组,每组各10只。后四组大鼠双侧海马CA1区注射p淀粉样蛋白25-35(Aβ25-35),假手术组双侧海马CA1区注射等体积的双蒸水,正常组不作任何处理。造模后补肾活血组以益智健脑浓缩液灌胃,补肾组以地黄饮子浓缩液灌胃,活血组以血府逐瘀汤浓缩液灌胃,其余3组大鼠予以等剂量双蒸水灌胃对照。造模后第21天及中药干预8周后,进行Morris水迷宫实验(Morris water maze test, MWM),最后采用半定量RT-PCR和免疫组化SP法测定大鼠海马组织中GSK-3β的表达的变化。
结果：
1.行为学检测结果：与其他治疗组相比,补肾活血组能有效的改善AD模型大鼠的学习、记忆能力。
2. RT-PCR及免疫组化结果：与其他治疗组相比,补肾活血组AD模型大鼠脑内GSK-3β相对表达明显下调。结论：
1.采用双侧海马注射Aβ25-35可建立成功的AD模型。
2.补肾活血法干预可有效改善AD模型大鼠的学习、记忆能力,优于补肾法或活血法。
3.补肾活血法干预后AD模型大鼠脑内GSK-3β水平明显下调,通过抑制GSK-3β的活性可降低Tau蛋白的异常高度磷酸化水平,防止Ap在神经元外大量沉积,且优于补肾法或活血法。
Objective:
To investigate the effects of Bushen Huoxue(Yizhi Jiannao granule) method、Bushen(Decoction of Rehmanniae granule) method and Huoxue (Xuefu Zhuyu granule) method decoction's on Alzheimer's disease model rats' learn and memory ability and the expression of GSK-3βin hippocampus, this work discusses partially the mechanism and preponderance of traditional Bushen Huoxue method to treat Alzheimer's disease.
Methods:
60 SD male rats were randomly divided into six groups by weight, normal group, sham controls group, model group, Bushen Huoxue group, Bushen group, Huoxue group, and various groups of 10 only. The last Four groups injects beta amyloid protein 25-35 (Aβ25-35) in bilateral hippocampal CA1 region, sham controls group injection steam water, normal group received no special treatment. The Bushen Huoxue group、Bushen group and Huoxue group are gavaged separately Yizhi Jiannao granule、Decoction of Rehmanniae granule and Xuefu Zhuyu granule, the other theree group are gavaged such equal dose double-distilled water. And after operation 21 days and Chinese medicine therapy eight weeks, carry out MWM(morris water maze test) behavior detection.Last,the hippocampal of rats were taken out to examine the expression of GSK-3βby semi-quantitative RT-PCR and immunohistochemistry SABC method.
Results:
1. Behavioral results:compared with other treatment group, the Bushen Huoxue group can effectively improve AD of rats' ability to learn and remember.
2. RT-PCR and immunohistochemical result:compared with other treatment group, the Bushen Huoxue group can effectively reduce the level of GSK-3βin the AD mice model.
Conclusion:
1. Bilateral hippocampal injection by Aβ25-35 can build successful AD model.
2. The Bushen Huoxue method can effectively improve learn and memory ability in the AD mice model, superior to Bushen group or Huoxue group.
3. After the Bushen Huoxue method therapy, the level of GSK-3pin the AD mice model were decreased, by inhibiting through GSK-3βactivity can reduce the phosphorylated proteins abnormal high level of Tau beta, prevent Aβin neurons, superior to Bushen group or Huoxue group.
研究补肾活血法(益智健脑颗粒)、补肾法(地黄饮子)和活血法(血府逐瘀汤)治疗对AD模型大鼠学习、记忆能力及海马区糖原合酶激酶-3β(GSK-3β)表达的影响,从而探讨补肾活血法治疗AD的优势及部分作用机制。
方法：
将60只雄性SD大鼠按体重随机分为6组,分别为正常组、假手术组、模型组、补肾活血组、补肾组、活血组,每组各10只。后四组大鼠双侧海马CA1区注射p淀粉样蛋白25-35(Aβ25-35),假手术组双侧海马CA1区注射等体积的双蒸水,正常组不作任何处理。造模后补肾活血组以益智健脑浓缩液灌胃,补肾组以地黄饮子浓缩液灌胃,活血组以血府逐瘀汤浓缩液灌胃,其余3组大鼠予以等剂量双蒸水灌胃对照。造模后第21天及中药干预8周后,进行Morris水迷宫实验(Morris water maze test, MWM),最后采用半定量RT-PCR和免疫组化SP法测定大鼠海马组织中GSK-3β的表达的变化。
结果：
1.行为学检测结果：与其他治疗组相比,补肾活血组能有效的改善AD模型大鼠的学习、记忆能力。
2. RT-PCR及免疫组化结果：与其他治疗组相比,补肾活血组AD模型大鼠脑内GSK-3β相对表达明显下调。结论：
1.采用双侧海马注射Aβ25-35可建立成功的AD模型。
2.补肾活血法干预可有效改善AD模型大鼠的学习、记忆能力,优于补肾法或活血法。
3.补肾活血法干预后AD模型大鼠脑内GSK-3β水平明显下调,通过抑制GSK-3β的活性可降低Tau蛋白的异常高度磷酸化水平,防止Ap在神经元外大量沉积,且优于补肾法或活血法。
Objective:
To investigate the effects of Bushen Huoxue(Yizhi Jiannao granule) method、Bushen(Decoction of Rehmanniae granule) method and Huoxue (Xuefu Zhuyu granule) method decoction's on Alzheimer's disease model rats' learn and memory ability and the expression of GSK-3βin hippocampus, this work discusses partially the mechanism and preponderance of traditional Bushen Huoxue method to treat Alzheimer's disease.
Methods:
60 SD male rats were randomly divided into six groups by weight, normal group, sham controls group, model group, Bushen Huoxue group, Bushen group, Huoxue group, and various groups of 10 only. The last Four groups injects beta amyloid protein 25-35 (Aβ25-35) in bilateral hippocampal CA1 region, sham controls group injection steam water, normal group received no special treatment. The Bushen Huoxue group、Bushen group and Huoxue group are gavaged separately Yizhi Jiannao granule、Decoction of Rehmanniae granule and Xuefu Zhuyu granule, the other theree group are gavaged such equal dose double-distilled water. And after operation 21 days and Chinese medicine therapy eight weeks, carry out MWM(morris water maze test) behavior detection.Last,the hippocampal of rats were taken out to examine the expression of GSK-3βby semi-quantitative RT-PCR and immunohistochemistry SABC method.
Results:
1. Behavioral results:compared with other treatment group, the Bushen Huoxue group can effectively improve AD of rats' ability to learn and remember.
2. RT-PCR and immunohistochemical result:compared with other treatment group, the Bushen Huoxue group can effectively reduce the level of GSK-3βin the AD mice model.
Conclusion:
1. Bilateral hippocampal injection by Aβ25-35 can build successful AD model.
2. The Bushen Huoxue method can effectively improve learn and memory ability in the AD mice model, superior to Bushen group or Huoxue group.
3. After the Bushen Huoxue method therapy, the level of GSK-3pin the AD mice model were decreased, by inhibiting through GSK-3βactivity can reduce the phosphorylated proteins abnormal high level of Tau beta, prevent Aβin neurons, superior to Bushen group or Huoxue group.
引文
[1]李焰生.中国防治认知功能障碍专家共识[J].中华老年医学杂志,2006(7)：485-487.
[2]Ferri C P, Prince M, Brayne C, et al. Global prevalence of dementia:a Delphi consensus study[J]. Lancet,2005,366(9503):2112-2117.
[3]Bamberger M E, Harris M E, Mcdonald D R, et al. A cell surface receptor complex for fibrillar beta-amyloid mediates microglial activation[J]. J Neurosci,2003,23(7):2665-2674.
[4]Pimplikar S W. Reassessing the amyloid cascade hypothesis of Alzheimer's disease[J]. Int J Biochem Cell Biol,2009,41(6):1261-1268.
[5]Masters C L, Simms G, Weinman N A, et al. Amyloid plaque core protein in Alzheimer disease and Down syndrome[J]. Proc Natl Acad Sci U S A,1985,82(12):4245-4249.
[6]Hernandez F, Lucas J J, Cuadros R, et al. GSK-3 dependent phosphoepitopes recognized by PHF-1 and AT-8 antibodies are present in different tau isoforms[J]. Neurobiol Aging,2003,24(8):1087-1094.
[7]Delcommenne M, Tan C, Gray V, et al. Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase[J]. Proc Natl Acad Sci U S A,1998, 95(19):11211-11216.
[8]Balaraman Y, Limaye A R, Levey A I, et al. Glycogen synthase kinase 3beta and Alzheimer's disease:pathophysiological and therapeutic significance[J]. Cell Mol Life Sci,2006,63(11):1226-1235.
[9]朱宏,董克礼,吴岳,等.补肾活血法对阿尔茨海默病患者认知功能改善的影响[J].中国老年学杂志,2010(11).
[10]吴岳,张婷.董克礼教授运用补肾活血法治疗中老年疾病经验介绍[J].中医药导报,2007(2)：23-24.
[11]张婷,董克礼.益智健脑颗粒对阿尔茨海默病患者脑脊液p淀粉样蛋白含量的影响[J].中国老年学杂志,2007(11)：1080-1081.
[12]王慧玲,董克礼,李广诚,等.益智健脑颗粒对快速老化小鼠SAMP8海马Pinl和HMGB1 mRNA表达的影响[J].中南大学学报(医学版),2009(1).
[13]龚翠兰,董克礼.益智健脑颗粒联合针灸对阿尔茨海默病大鼠学习记忆的影响[J].湖南中医药大学学报,2009(5).
[14]包新民,舒斯云著.大鼠立体定位图谱[M].人民卫生出版社,1991.
[15]凌雁武,黄瑞雅.老年性痴呆动物模型的研究进展[J].右江医学,2008(6)：736-740.
[16]魏小龙,张永祥.老年性痴呆动物模型研究进展[J].中国药理学通报,2000(4)：372-376.
[17]贾建平.神经病学[M].人民卫生出版社,2008：473.
[18]周建军,张馨,徐运.雌激素对老年性痴呆的防治作用及其分子机制的新进展[J].国外医学.神经病学神经外科学分册,2005(1)：99-102.
[19]Fox N C, Black R S, Gilman S, et al. Effects of Abeta immunization (AN 1792) on MRI measures of cerebral volume in Alzheimer disease[J]. Neurology, 2005,64(9):1563-1572.
[20]Yamada M, Chiba T, Sasabe J, et al. Implanted cannula-mediated repetitive administration of Abeta25-35 into the mouse cerebral ventricle effectively impairs spatial working memory[J]. Behav Brain Res,2005,164(2):139-146.
[21]吴树亮,金连弘,李竹琴,等.阿尔茨海默氏病动物模型的建立[J].解剖科学进展,2004(2).
[22]Pike C J, Burdick D, Walencewicz A J, et al. Neurodegeneration induced by beta-amyloid peptides in vitro:the role of peptide assembly state[J]. J Neurosci,1993,13(4):1676-1687.
[23]于洋,李萌,侯艳宁,等.Aβ_(25-35)对原代培养大鼠皮质神经元的损伤作用[J].华北国防医药,2010(4)：301-304.
[24]王德生.老年性痴呆[M].人民卫生出版社,2001：523.
[25]牛英才,兴桂华,林春荣,等.Ap海马注射对大鼠学习记忆影响及加减地黄饮子的干预作用[J].辽宁中医杂志,2007(7).
[26]贺石林,陈修.医学科研方法导论[M].北京：人民卫生出版社,1998,62-64
[27]尚玉昌.动物行为学[M].北京大学出版社,2005：384.
[28]朱宏,董克礼,吴岳,等.针刺对阿尔茨海默病患者异构前列腺素的影响[J].中国针灸,2010(1).
[29]Squire L R. Memory and brain systems:1969-2009[J]. J Neurosci,2009, 29(41):12711-12716.
[30]朱荣.中医对阿尔茨海默病的病机认识与治疗方法探讨[J].山西中医,2008(10)：1-3.
[31]朱宏,董克礼,吴岳,等.补肾活血法对阿尔茨海默病患者认知功能改善的影响[J]..中国老年学杂志,2010(11).
[32]董桂英,赵世珂,郭立华.补肾活血法治疗老年期痴呆临床观察[J].：15-16.
[33]彭勇,张荣华,蔡宇,等.补肾活血法延缓衰老的机理探讨[J].陕西中医,2004(2)：142-144.
[34]龙斯玥,周德生.补肾活血法治疗老年性痴呆的研究进展[J].世界中医药,2008(5)：319-320.
[35]郑亿,张贺,王文凯.地黄饮予子防老年痴呆的行为学实验研究[J].黑龙江医学,2008(2)：118-120.
[36]牛英才,董妙先,兴桂华,等.Aβ1-40脑内注射对大鼠海马AchE和NOS 1表达的影响及加减地黄饮子的干预作用[J].中药药理与临床,2007(5)：149-151.
[37]谢宁,邹纯朴,周妍妍,等.地黄饮子脑脊液对离体神经元AD模型自由基损伤的保护作用及其相关酶的影响[J].中医药信息,2003(6)：59-61.
[38]黄云飞,王兴华,屈新宇,等.血府逐瘀汤药理研究进展[J].黑龙江医药,1998(3)：164-165.
[39]王学洋,丁涛.血府逐瘀汤药理学研究进展[J].中国社区医师(综合版),2007(12)：8.
[40]Arriagada P V, Growdon J H, Hedley-Whyte E T, et al. Neurofibrillary tangles but not senile plaques parallel duration and severity of Alzheimer's disease[J]. Neurology,1992,42(3 Pt 1):631-639.
[41]Gong C X, Liu F, Grundke-Iqbal Ⅰ, et al. Post-translational modifications of tau protein in Alzheimer's disease[J]. J Neural Transm,2005,112(6):813-838.
[42]Boonen R A, van Tijn P, Zivkovic D. Wnt signaling in Alzheimer's disease: up or down, that is the question[J]. Ageing Res Rev,2009,8(2):71-82.
[43]Rockenstein E, Torrance M, Adame A, et al. Neuroprotective effects of regulators of the glycogen synthase kinase-3beta signaling pathway in a transgenic model of Alzheimer's disease are associated with reduced amyloid precursor protein phosphorylation[J]. J Neurosci,2007,27(8):1981-1991.
[44]Hu S, Begum A N, Jones M R, et al. GSK3 inhibitors show benefits in an Alzheimer's disease (AD) model of neurodegeneration but adverse effects in control animals[J]. Neurobiol Dis,2009,33(2):193-206.
[45]Hooper C, Killick R, Lovestone S. The GSK3 hypothesis of Alzheimer's disease[J]. J Neurochem,2008,104(6):1433-1439.
[46]Loo D T, Copani A, Pike C J, et al. Apoptosis is induced by beta-amyloid in cultured central nervous system neurons[J]. Proc Natl Acad Sci U S A,1993,90(17):7951-7955.
[47]Defelice F G, Ferreira S T. Physiopathological modulators of amyloid aggregation and novel pharmacological approaches in Alzheimer's disease[J]. An Acad Bras Cienc,2002,74(2):265-284.
[48]Roberson E D, Scearce-Levie K, Palop J J, et al. Reducing endogenous tau ameliorates amyloid beta-induced deficits in an Alzheimer's disease mouse model[J]. Science,2007,316(5825):750-754.
[49]陈林,姜招峰.糖原合酶激酶-3β在淀粉样蛋白质前体代谢中的作用[J].生命的化学,2009(2)：227-230.
[50]Alonso A C, Zaidi T, Grundke-Iqbal I, et al. Role of abnormally phosphorylat-ed tau in the breakdown of microtubules in Alzheimer disease[J]. Proc Natl Acad Sci U S A,1994,91(12):5562-5566.
[51]Iqbal K, Alonso A C, Chen S, et al. Tau pathology in Alzheimer disease and other tauopathies[J]. Biochim Biophys Acta,2005,1739(2-3):198-210.
[52]Caricasole A, Copani A, Caraci F, et al. Induction of Dickkopf-1, a negative modulator of the Wnt pathway, is associated with neuronal degeneration in Alzheimer's brain[J]. J Neurosci,2004,24(26):6021-6027.
[53]Baki L, Shioi J, Wen P, et al. PS1 activates PI3K thus inhibiting GSK-3 activity and tau overphosphorylation:effects of FAD mutations [J]. EMBO J,2004,23(13):2586-2596.
[1]刘晶,宋光,张男.阿尔茨海默症与糖原合酶激酶-3的相关联系[J].人人健康(医学导刊),2008(5)：81-83.
[2]Hooper C, Killick R, Lovestone S. The GSK3 hypothesis of Alzheimer's disease[J]. J Neurochem,2008,104(6):1433-1439.
[3]Mandelkow E M, Drewes G, Biernat J, et al. Glycogen synthase kinase-3 and the Alzheimer-like state of microtubule-associated protein tau[J]. FEBS Lett,1992,314(3):315-321.
[4]Balaraman Y, Limaye A R, Levey A I, et al. Glycogen synthase kinase 3beta and Alzheimer's disease:pathophysiological and therapeutic significance[J]. Cell Mol Life Sci,2006,63(11):1226-1235.
[5]Boonen R A, van Tijn P, Zivkovic D. Wnt signaling in Alzheimer's disease: up or down, that is the question[J]. Ageing Res Rev,2009,8(2):71-82.
[6]Rockenstein E, Torrance M, Adame A, et al. Neuroprotective effects of regulators of the glycogen synthase kinase-3beta signaling pathway in a transgenic model of Alzheimer's disease are associated with reduced amyloid precursor protein phosphorylation[J]. J Neurosci,2007,27(8):1981-1991.
[7]Hoshi M, Takashima A, Noguchi K, et al. Regulation of mitochondrial pyruvate dehydrogenase activity by tau protein kinase I/glycogen synthase kinase 3beta in brain[J]. Proc Natl Acad Sci U S A,1996,93(7):2719-2723.
[8]Yamaguchi H, Ishiguro K, Uchida T, et al. Preferential labeling of Alzheimer neurofibrillary tangles with antisera for tau protein kinase (TPK) I/glycogen synthase kinase-3 beta and cyclin-dependent kinase 5, a component of TPK II[J]. Acta Neuropathol,1996,92(3):232-241.
[9]Pei J J, Braak E, Braak H, et al. Distribution of active glycogen synthase kinase 3beta (GSK-3beta) in brains staged for Alzheimer disease neurofibrillary changes[J]. J Neuropathol Exp Neurol,1999,58(9):1010-1019.
[10]Leroy K, Yilmaz Z, Brion J P. Increased level of active GSK-3beta in Alzheimer's disease and accumulation in argyrophilic grains and in neurones at different stages of neurofibrillary degeneration[J]. Neuropathol Appl Neurobiol,2007,33(1):43-55.
[11]Hye A, Kerr F, Archer N, et al. Glycogen synthase kinase-3 is increased in white cells early in Alzheimer's disease[J]. Neurosci Lett,2005,373(1):1-4.
[12]Mateo I, Infante J, Llorca J, et al. Association between glycogen synthase kinase-3beta genetic polymorphism and late-onset Alzheimer's disease[J]. Dement Geriatr Cogn Disord,2006,21(4):228-232.
[13]Swatton J E, Sellers L A, Faull R L, et al. Increased MAP kinase activity in Alzheimer's and Down syndrome but not in schizophrenia human brain[J]. Eur J Neurosci,2004,19(10):2711-2719.
[14]De Ferrari G V, Papassotiropoulos A, Biechele T, et al. Common genetic variation within the low-density lipoprotein receptor-related protein 6 and late-onset Alzheimer's disease[J]. Proc Natl Acad Sci U S A,2007, 104(22):9434-9439.
[15]Hamilton G, Proitsi P, Jehu L, et al. Candidate gene association study of insulin signaling genes and Alzheimer's disease:evidence for SOS2, PCK1, and PPARgamma as susceptibility loci[J]. Am J Med Genet B Neuropsychiatr Genet,2007,144B(4):508-516.
[16]Hanger D P, Hughes K, Woodgett J R, et al. Glycogen synthase kinase-3 induces Alzheimer's disease-like phosphorylation of tau:generation of paired helical filament epitopes and neuronal localisation of the kinase[J]. Neurosci Lett,1992,147(1):58-62.
[17]Cho J H, Johnson G V. Glycogen synthase kinase 3beta phosphorylates tau at both primed and unprimed sites. Differential impact on microtubule binding[J]. J Biol Chem,2003,278(1):187-193.
[18]Lucas J J, Hernandez F, Gomez-Ramos P, et al. Decreased nuclear beta-catenin, tau hyperphosphorylation and neurodegeneration in GSK-3beta conditional transgenic mice[J]. EMBO J,2001,20(1-2):27-39.
[19]Mudher A, Shepherd D, Newman T A, et al. GSK-3beta inhibition reverses axonal transport defects and behavioural phenotypes in Drosophila[J]. Mol Psychiatry,2004,9(5):522-530.
[20]Caricasole A, Copani A, Caraci F, et al. Induction of Dickkopf-1, a negative modulator of the Wnt pathway, is associated with neuronal degeneration in Alzheimer's brain[J]. J Neurosci,2004,24(26):6021-6027.
[21]Baki L, Shioi J, Wen P, et al. PS1 activates PI3K thus inhibiting GSK-3 activity and tau overphosphorylation:effects of FAD mutations[J]. EMBO J,2004,23(13):2586-2596.
[22]Sun X, Sato S, Murayama O, et al. Lithium inhibits amyloid secretion in COS7 cells transfected with amyloid precursor protein C100[J]. Neurosci Lett,2002,321(1-2):61-64.
[23]Alvarez G, Munoz-Montano J R, Satrustegui J, et al. Lithium protects cultured neurons against beta-amyloid-induced neurodegeneration[J]. FEBS Lett,1999,453(3):260-264.
[24]Zhao L, Teter B, Morihara T, et al. Insulin-degrading enzyme as a downstre-am target of insulin receptor signaling cascade:implications for Alzheimer's disease intervention[J]. J Neurosci,2004,24(49):11120-11126.
[25]Reger M A, Watson G S, Green P S, et al. Intranasal insulin improves cognition and modulates beta-amyloid in early AD[J]. Neurology, 2008,70(6):440-448.
[26]Welsh G I, Proud C G. Glycogen synthase kinase-3 is rapidly inactivated in response to insulin and phosphorylates eukaryotic initiation factor eIF-2B[J]. Biochem J,1993,294 (Pt 3):625-629.
[27]Macaulay K, Doble B W, Patel S, et al. Glycogen synthase kinase 3alpha-specific regulation of murine hepatic glycogen metabolism[J]. Cell Metab,2007,6(4):329-337.
[28]Lizcano J M, Alessi D R. The insulin signalling pathway[J]. Curr Biol,2002,12(7):R236-R238.
[29]Doble B W, Woodgett J R. GSK-3:tricks of the trade for a multi-tasking kinase[J]. J Cell Sci,2003,116(Pt 7):1175-1186.
[30]Bhat R V, Shanley J, Correll M P, et al. Regulation and localization of tyrosine216 phosphorylation of glycogen synthase kinase-3beta in cellular and animal models of neuronal degeneration[J]. Proc Natl Acad Sci U S A,2000,97(20):11074-11079.
[31]Nusse R. Wnt signaling in disease and in development[J]. Cell Res,2005, 15(1):28-32.
[32]Doble B W, Patel S, Wood G A, et al. Functional redundancy of GSK-3alpha and GSK-3beta in Wnt/beta-catenin signaling shown by using an allelic series of embryonic stem cell lines[J]. Dev Cell,2007,12(6):957-971.
[33]Hooper C, Markevich V, Plattner F, et al. Glycogen synthase kinase-3 inhibition is integral to long-term potentiation[J]. Eur J Neurosci,2007, 25(1):81-86.
[34]Peineau S, Taghibiglou C, Bradley C, et al. LTP inhibits LTD in the hippocampus via regulation of GSK3beta[J]. Neuron,2007,53(5):703-717.
[35]Cole A R, Knebel A, Morrice N A, et al. GSK-3 phosphorylation of the Alzheimer epitope within collapsin response mediator proteins regulates axon elongation in primary neurons[J]. J Biol Chem,2004,279(48):50176-50180.
[36]Hansen T O, Rehfeld J F, Nielsen F C. GSK-3beta reduces cAMP-induced cholecystokinin gene expression by inhibiting CREB binding[J]. Neuroreport,2004,15(5):841-845.
[37]Koivisto L, Hakkinen L, Matsumoto K, et al. Glycogen synthase kinase-3 regulates cytoskeleton and translocation of Racl in long cellular extensions of human keratinocytes[J]. Exp Cell Res,2004,293(1):68-80.
[38]Mcgeer P L, Mcgeer E G. Anti-inflammatory drugs in the fight against Alzheimer's disease[J]. Ann N Y Acad Sci,1996,777:213-220.
[39]Schenk D B, Seubert P, Grundman M, et al. A beta immunotherapy:Lessons learned for potential treatment of Alzheimer's disease[J]. Neurodegener Dis,2005,2(5):255-260.
[40]Rodionova E, Conzelmann M, Maraskovsky E, et al. GSK-3 mediates differentiation and activation of proinflammatory dendritic cells[J]. Blood,2007,109(4):1584-1592.
[41]Rockenstein E, Torrance M, Adame A, et al. Neuroprotective effects of regulators of the glycogen synthase kinase-3beta signaling pathway in a transgenic model of Alzheimer's disease are associated with reduced amyloid precursor protein phosphorylation[J]. J Neurosci,2007,27(8):1981-1991.
[2]Ferri C P, Prince M, Brayne C, et al. Global prevalence of dementia:a Delphi consensus study[J]. Lancet,2005,366(9503):2112-2117.
[3]Bamberger M E, Harris M E, Mcdonald D R, et al. A cell surface receptor complex for fibrillar beta-amyloid mediates microglial activation[J]. J Neurosci,2003,23(7):2665-2674.
[4]Pimplikar S W. Reassessing the amyloid cascade hypothesis of Alzheimer's disease[J]. Int J Biochem Cell Biol,2009,41(6):1261-1268.
[5]Masters C L, Simms G, Weinman N A, et al. Amyloid plaque core protein in Alzheimer disease and Down syndrome[J]. Proc Natl Acad Sci U S A,1985,82(12):4245-4249.
[6]Hernandez F, Lucas J J, Cuadros R, et al. GSK-3 dependent phosphoepitopes recognized by PHF-1 and AT-8 antibodies are present in different tau isoforms[J]. Neurobiol Aging,2003,24(8):1087-1094.
[7]Delcommenne M, Tan C, Gray V, et al. Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase[J]. Proc Natl Acad Sci U S A,1998, 95(19):11211-11216.
[8]Balaraman Y, Limaye A R, Levey A I, et al. Glycogen synthase kinase 3beta and Alzheimer's disease:pathophysiological and therapeutic significance[J]. Cell Mol Life Sci,2006,63(11):1226-1235.
[9]朱宏,董克礼,吴岳,等.补肾活血法对阿尔茨海默病患者认知功能改善的影响[J].中国老年学杂志,2010(11).
[10]吴岳,张婷.董克礼教授运用补肾活血法治疗中老年疾病经验介绍[J].中医药导报,2007(2)：23-24.
[11]张婷,董克礼.益智健脑颗粒对阿尔茨海默病患者脑脊液p淀粉样蛋白含量的影响[J].中国老年学杂志,2007(11)：1080-1081.
[12]王慧玲,董克礼,李广诚,等.益智健脑颗粒对快速老化小鼠SAMP8海马Pinl和HMGB1 mRNA表达的影响[J].中南大学学报(医学版),2009(1).
[13]龚翠兰,董克礼.益智健脑颗粒联合针灸对阿尔茨海默病大鼠学习记忆的影响[J].湖南中医药大学学报,2009(5).
[14]包新民,舒斯云著.大鼠立体定位图谱[M].人民卫生出版社,1991.
[15]凌雁武,黄瑞雅.老年性痴呆动物模型的研究进展[J].右江医学,2008(6)：736-740.
[16]魏小龙,张永祥.老年性痴呆动物模型研究进展[J].中国药理学通报,2000(4)：372-376.
[17]贾建平.神经病学[M].人民卫生出版社,2008：473.
[18]周建军,张馨,徐运.雌激素对老年性痴呆的防治作用及其分子机制的新进展[J].国外医学.神经病学神经外科学分册,2005(1)：99-102.
[19]Fox N C, Black R S, Gilman S, et al. Effects of Abeta immunization (AN 1792) on MRI measures of cerebral volume in Alzheimer disease[J]. Neurology, 2005,64(9):1563-1572.
[20]Yamada M, Chiba T, Sasabe J, et al. Implanted cannula-mediated repetitive administration of Abeta25-35 into the mouse cerebral ventricle effectively impairs spatial working memory[J]. Behav Brain Res,2005,164(2):139-146.
[21]吴树亮,金连弘,李竹琴,等.阿尔茨海默氏病动物模型的建立[J].解剖科学进展,2004(2).
[22]Pike C J, Burdick D, Walencewicz A J, et al. Neurodegeneration induced by beta-amyloid peptides in vitro:the role of peptide assembly state[J]. J Neurosci,1993,13(4):1676-1687.
[23]于洋,李萌,侯艳宁,等.Aβ_(25-35)对原代培养大鼠皮质神经元的损伤作用[J].华北国防医药,2010(4)：301-304.
[24]王德生.老年性痴呆[M].人民卫生出版社,2001：523.
[25]牛英才,兴桂华,林春荣,等.Ap海马注射对大鼠学习记忆影响及加减地黄饮子的干预作用[J].辽宁中医杂志,2007(7).
[26]贺石林,陈修.医学科研方法导论[M].北京：人民卫生出版社,1998,62-64
[27]尚玉昌.动物行为学[M].北京大学出版社,2005：384.
[28]朱宏,董克礼,吴岳,等.针刺对阿尔茨海默病患者异构前列腺素的影响[J].中国针灸,2010(1).
[29]Squire L R. Memory and brain systems:1969-2009[J]. J Neurosci,2009, 29(41):12711-12716.
[30]朱荣.中医对阿尔茨海默病的病机认识与治疗方法探讨[J].山西中医,2008(10)：1-3.
[31]朱宏,董克礼,吴岳,等.补肾活血法对阿尔茨海默病患者认知功能改善的影响[J]..中国老年学杂志,2010(11).
[32]董桂英,赵世珂,郭立华.补肾活血法治疗老年期痴呆临床观察[J].：15-16.
[33]彭勇,张荣华,蔡宇,等.补肾活血法延缓衰老的机理探讨[J].陕西中医,2004(2)：142-144.
[34]龙斯玥,周德生.补肾活血法治疗老年性痴呆的研究进展[J].世界中医药,2008(5)：319-320.
[35]郑亿,张贺,王文凯.地黄饮予子防老年痴呆的行为学实验研究[J].黑龙江医学,2008(2)：118-120.
[36]牛英才,董妙先,兴桂华,等.Aβ1-40脑内注射对大鼠海马AchE和NOS 1表达的影响及加减地黄饮子的干预作用[J].中药药理与临床,2007(5)：149-151.
[37]谢宁,邹纯朴,周妍妍,等.地黄饮子脑脊液对离体神经元AD模型自由基损伤的保护作用及其相关酶的影响[J].中医药信息,2003(6)：59-61.
[38]黄云飞,王兴华,屈新宇,等.血府逐瘀汤药理研究进展[J].黑龙江医药,1998(3)：164-165.
[39]王学洋,丁涛.血府逐瘀汤药理学研究进展[J].中国社区医师(综合版),2007(12)：8.
[40]Arriagada P V, Growdon J H, Hedley-Whyte E T, et al. Neurofibrillary tangles but not senile plaques parallel duration and severity of Alzheimer's disease[J]. Neurology,1992,42(3 Pt 1):631-639.
[41]Gong C X, Liu F, Grundke-Iqbal Ⅰ, et al. Post-translational modifications of tau protein in Alzheimer's disease[J]. J Neural Transm,2005,112(6):813-838.
[42]Boonen R A, van Tijn P, Zivkovic D. Wnt signaling in Alzheimer's disease: up or down, that is the question[J]. Ageing Res Rev,2009,8(2):71-82.
[43]Rockenstein E, Torrance M, Adame A, et al. Neuroprotective effects of regulators of the glycogen synthase kinase-3beta signaling pathway in a transgenic model of Alzheimer's disease are associated with reduced amyloid precursor protein phosphorylation[J]. J Neurosci,2007,27(8):1981-1991.
[44]Hu S, Begum A N, Jones M R, et al. GSK3 inhibitors show benefits in an Alzheimer's disease (AD) model of neurodegeneration but adverse effects in control animals[J]. Neurobiol Dis,2009,33(2):193-206.
[45]Hooper C, Killick R, Lovestone S. The GSK3 hypothesis of Alzheimer's disease[J]. J Neurochem,2008,104(6):1433-1439.
[46]Loo D T, Copani A, Pike C J, et al. Apoptosis is induced by beta-amyloid in cultured central nervous system neurons[J]. Proc Natl Acad Sci U S A,1993,90(17):7951-7955.
[47]Defelice F G, Ferreira S T. Physiopathological modulators of amyloid aggregation and novel pharmacological approaches in Alzheimer's disease[J]. An Acad Bras Cienc,2002,74(2):265-284.
[48]Roberson E D, Scearce-Levie K, Palop J J, et al. Reducing endogenous tau ameliorates amyloid beta-induced deficits in an Alzheimer's disease mouse model[J]. Science,2007,316(5825):750-754.
[49]陈林,姜招峰.糖原合酶激酶-3β在淀粉样蛋白质前体代谢中的作用[J].生命的化学,2009(2)：227-230.
[50]Alonso A C, Zaidi T, Grundke-Iqbal I, et al. Role of abnormally phosphorylat-ed tau in the breakdown of microtubules in Alzheimer disease[J]. Proc Natl Acad Sci U S A,1994,91(12):5562-5566.
[51]Iqbal K, Alonso A C, Chen S, et al. Tau pathology in Alzheimer disease and other tauopathies[J]. Biochim Biophys Acta,2005,1739(2-3):198-210.
[52]Caricasole A, Copani A, Caraci F, et al. Induction of Dickkopf-1, a negative modulator of the Wnt pathway, is associated with neuronal degeneration in Alzheimer's brain[J]. J Neurosci,2004,24(26):6021-6027.
[53]Baki L, Shioi J, Wen P, et al. PS1 activates PI3K thus inhibiting GSK-3 activity and tau overphosphorylation:effects of FAD mutations [J]. EMBO J,2004,23(13):2586-2596.
[1]刘晶,宋光,张男.阿尔茨海默症与糖原合酶激酶-3的相关联系[J].人人健康(医学导刊),2008(5)：81-83.
[2]Hooper C, Killick R, Lovestone S. The GSK3 hypothesis of Alzheimer's disease[J]. J Neurochem,2008,104(6):1433-1439.
[3]Mandelkow E M, Drewes G, Biernat J, et al. Glycogen synthase kinase-3 and the Alzheimer-like state of microtubule-associated protein tau[J]. FEBS Lett,1992,314(3):315-321.
[4]Balaraman Y, Limaye A R, Levey A I, et al. Glycogen synthase kinase 3beta and Alzheimer's disease:pathophysiological and therapeutic significance[J]. Cell Mol Life Sci,2006,63(11):1226-1235.
[5]Boonen R A, van Tijn P, Zivkovic D. Wnt signaling in Alzheimer's disease: up or down, that is the question[J]. Ageing Res Rev,2009,8(2):71-82.
[6]Rockenstein E, Torrance M, Adame A, et al. Neuroprotective effects of regulators of the glycogen synthase kinase-3beta signaling pathway in a transgenic model of Alzheimer's disease are associated with reduced amyloid precursor protein phosphorylation[J]. J Neurosci,2007,27(8):1981-1991.
[7]Hoshi M, Takashima A, Noguchi K, et al. Regulation of mitochondrial pyruvate dehydrogenase activity by tau protein kinase I/glycogen synthase kinase 3beta in brain[J]. Proc Natl Acad Sci U S A,1996,93(7):2719-2723.
[8]Yamaguchi H, Ishiguro K, Uchida T, et al. Preferential labeling of Alzheimer neurofibrillary tangles with antisera for tau protein kinase (TPK) I/glycogen synthase kinase-3 beta and cyclin-dependent kinase 5, a component of TPK II[J]. Acta Neuropathol,1996,92(3):232-241.
[9]Pei J J, Braak E, Braak H, et al. Distribution of active glycogen synthase kinase 3beta (GSK-3beta) in brains staged for Alzheimer disease neurofibrillary changes[J]. J Neuropathol Exp Neurol,1999,58(9):1010-1019.
[10]Leroy K, Yilmaz Z, Brion J P. Increased level of active GSK-3beta in Alzheimer's disease and accumulation in argyrophilic grains and in neurones at different stages of neurofibrillary degeneration[J]. Neuropathol Appl Neurobiol,2007,33(1):43-55.
[11]Hye A, Kerr F, Archer N, et al. Glycogen synthase kinase-3 is increased in white cells early in Alzheimer's disease[J]. Neurosci Lett,2005,373(1):1-4.
[12]Mateo I, Infante J, Llorca J, et al. Association between glycogen synthase kinase-3beta genetic polymorphism and late-onset Alzheimer's disease[J]. Dement Geriatr Cogn Disord,2006,21(4):228-232.
[13]Swatton J E, Sellers L A, Faull R L, et al. Increased MAP kinase activity in Alzheimer's and Down syndrome but not in schizophrenia human brain[J]. Eur J Neurosci,2004,19(10):2711-2719.
[14]De Ferrari G V, Papassotiropoulos A, Biechele T, et al. Common genetic variation within the low-density lipoprotein receptor-related protein 6 and late-onset Alzheimer's disease[J]. Proc Natl Acad Sci U S A,2007, 104(22):9434-9439.
[15]Hamilton G, Proitsi P, Jehu L, et al. Candidate gene association study of insulin signaling genes and Alzheimer's disease:evidence for SOS2, PCK1, and PPARgamma as susceptibility loci[J]. Am J Med Genet B Neuropsychiatr Genet,2007,144B(4):508-516.
[16]Hanger D P, Hughes K, Woodgett J R, et al. Glycogen synthase kinase-3 induces Alzheimer's disease-like phosphorylation of tau:generation of paired helical filament epitopes and neuronal localisation of the kinase[J]. Neurosci Lett,1992,147(1):58-62.
[17]Cho J H, Johnson G V. Glycogen synthase kinase 3beta phosphorylates tau at both primed and unprimed sites. Differential impact on microtubule binding[J]. J Biol Chem,2003,278(1):187-193.
[18]Lucas J J, Hernandez F, Gomez-Ramos P, et al. Decreased nuclear beta-catenin, tau hyperphosphorylation and neurodegeneration in GSK-3beta conditional transgenic mice[J]. EMBO J,2001,20(1-2):27-39.
[19]Mudher A, Shepherd D, Newman T A, et al. GSK-3beta inhibition reverses axonal transport defects and behavioural phenotypes in Drosophila[J]. Mol Psychiatry,2004,9(5):522-530.
[20]Caricasole A, Copani A, Caraci F, et al. Induction of Dickkopf-1, a negative modulator of the Wnt pathway, is associated with neuronal degeneration in Alzheimer's brain[J]. J Neurosci,2004,24(26):6021-6027.
[21]Baki L, Shioi J, Wen P, et al. PS1 activates PI3K thus inhibiting GSK-3 activity and tau overphosphorylation:effects of FAD mutations[J]. EMBO J,2004,23(13):2586-2596.
[22]Sun X, Sato S, Murayama O, et al. Lithium inhibits amyloid secretion in COS7 cells transfected with amyloid precursor protein C100[J]. Neurosci Lett,2002,321(1-2):61-64.
[23]Alvarez G, Munoz-Montano J R, Satrustegui J, et al. Lithium protects cultured neurons against beta-amyloid-induced neurodegeneration[J]. FEBS Lett,1999,453(3):260-264.
[24]Zhao L, Teter B, Morihara T, et al. Insulin-degrading enzyme as a downstre-am target of insulin receptor signaling cascade:implications for Alzheimer's disease intervention[J]. J Neurosci,2004,24(49):11120-11126.
[25]Reger M A, Watson G S, Green P S, et al. Intranasal insulin improves cognition and modulates beta-amyloid in early AD[J]. Neurology, 2008,70(6):440-448.
[26]Welsh G I, Proud C G. Glycogen synthase kinase-3 is rapidly inactivated in response to insulin and phosphorylates eukaryotic initiation factor eIF-2B[J]. Biochem J,1993,294 (Pt 3):625-629.
[27]Macaulay K, Doble B W, Patel S, et al. Glycogen synthase kinase 3alpha-specific regulation of murine hepatic glycogen metabolism[J]. Cell Metab,2007,6(4):329-337.
[28]Lizcano J M, Alessi D R. The insulin signalling pathway[J]. Curr Biol,2002,12(7):R236-R238.
[29]Doble B W, Woodgett J R. GSK-3:tricks of the trade for a multi-tasking kinase[J]. J Cell Sci,2003,116(Pt 7):1175-1186.
[30]Bhat R V, Shanley J, Correll M P, et al. Regulation and localization of tyrosine216 phosphorylation of glycogen synthase kinase-3beta in cellular and animal models of neuronal degeneration[J]. Proc Natl Acad Sci U S A,2000,97(20):11074-11079.
[31]Nusse R. Wnt signaling in disease and in development[J]. Cell Res,2005, 15(1):28-32.
[32]Doble B W, Patel S, Wood G A, et al. Functional redundancy of GSK-3alpha and GSK-3beta in Wnt/beta-catenin signaling shown by using an allelic series of embryonic stem cell lines[J]. Dev Cell,2007,12(6):957-971.
[33]Hooper C, Markevich V, Plattner F, et al. Glycogen synthase kinase-3 inhibition is integral to long-term potentiation[J]. Eur J Neurosci,2007, 25(1):81-86.
[34]Peineau S, Taghibiglou C, Bradley C, et al. LTP inhibits LTD in the hippocampus via regulation of GSK3beta[J]. Neuron,2007,53(5):703-717.
[35]Cole A R, Knebel A, Morrice N A, et al. GSK-3 phosphorylation of the Alzheimer epitope within collapsin response mediator proteins regulates axon elongation in primary neurons[J]. J Biol Chem,2004,279(48):50176-50180.
[36]Hansen T O, Rehfeld J F, Nielsen F C. GSK-3beta reduces cAMP-induced cholecystokinin gene expression by inhibiting CREB binding[J]. Neuroreport,2004,15(5):841-845.
[37]Koivisto L, Hakkinen L, Matsumoto K, et al. Glycogen synthase kinase-3 regulates cytoskeleton and translocation of Racl in long cellular extensions of human keratinocytes[J]. Exp Cell Res,2004,293(1):68-80.
[38]Mcgeer P L, Mcgeer E G. Anti-inflammatory drugs in the fight against Alzheimer's disease[J]. Ann N Y Acad Sci,1996,777:213-220.
[39]Schenk D B, Seubert P, Grundman M, et al. A beta immunotherapy:Lessons learned for potential treatment of Alzheimer's disease[J]. Neurodegener Dis,2005,2(5):255-260.
[40]Rodionova E, Conzelmann M, Maraskovsky E, et al. GSK-3 mediates differentiation and activation of proinflammatory dendritic cells[J]. Blood,2007,109(4):1584-1592.
[41]Rockenstein E, Torrance M, Adame A, et al. Neuroprotective effects of regulators of the glycogen synthase kinase-3beta signaling pathway in a transgenic model of Alzheimer's disease are associated with reduced amyloid precursor protein phosphorylation[J]. J Neurosci,2007,27(8):1981-1991.