17α雌二醇对APP/PS1 N_2a基因转染细胞Aβ生成的影响及其机制的研究
摘要
目的：探讨17α雌二醇(E2α)对稳定表达人APP/PS1基因的神经母细胞瘤细胞株(Neuro-2a) (APP/PS1 N2a)β-淀粉样蛋白(Aβ)产生的影响及可能机制。
方法：将APP/PS1N2a细胞用10 nM E2α预处理2天,更换培养液后,分别进行如下处理：(1)加入不同浓度E2α(25nM,50nM,100nM,200nM)和溶媒(对照组),用Western-blot检测APP/PS1N2a细胞内和细胞外的Ap蛋白水平及细胞BACE1蛋白水平；(2)加入100 nM E2α、20 mM葡萄糖和5 mU葡萄糖氧化酶(glucose oxidase, GOX),在培养液中GOX可以分解G并产生H202,对细胞造成氧化损伤。24h后,进行DHE染色观察APP/PS1 N2a细胞中活性氧(ROS)含量的变化。
结果：不同浓度E2α(25nM,50nM,100nM,200nM)处理后,细胞外Ap蛋白水平明显减少,差异具有统计学意义(P<0.05)；而细胞内Ap蛋白水平无明显变化(P>0.05)；细胞BACE1的表达水平无明显变化(P>0.05)；E2α可以减少G/GOX所致APP/PS1 N2a细胞ROS的产生。
结论：E2α可以减少细胞外Ap的产生,其机制可能与其抗氧化损伤有关。
Objective To investigate the effects of 17a-estradiol (E2a) on the release ofβ-amyloid protein (Aβ) with the transgenic cell line APP/PS 1 N2a, and discuss the potential mechanism of such effects.
Methods APP/PS 1 N2a cell lines were pretreated with 10 nM E2a for 2 days, then changed the culture medium, and treated with different methods:(1) cell lines were treated with either E2a (25nM,50nM, 100nM,200nM) or vehicle, The protein level of Aβand BACE1 both intracellular and extracellular of APP/PS 1 N2a cells were detected by Western blot. (2) cell lines were treated with 100nM E2a,20 mM glucose and 5 mU glucose oxidase(GOX), followed by exposure to the glucose oxidase (G/Gox) to induce oxidative stress. After 24 hours, the change of the ROS contents in APP/PS 1 N2a cells was observed for DHE.
Results Pretreatment of APP/PS 1 N2a cells with different concentration (25nM,50nM, 100nM,200nM) E2a significantly reduced extracellular levels of AP(P<0.05). There was no change both in the intracellular levels of Aβand the levels of BACE1(P>0.05). E2a can reduce the ROS produced by APP/PS 1 N2a cells after G/Gox treatment.
Conclusion E2a can reduce the production of extracellular Aβ,its mechanism may be related to the oxidation damage.
方法：将APP/PS1N2a细胞用10 nM E2α预处理2天,更换培养液后,分别进行如下处理：(1)加入不同浓度E2α(25nM,50nM,100nM,200nM)和溶媒(对照组),用Western-blot检测APP/PS1N2a细胞内和细胞外的Ap蛋白水平及细胞BACE1蛋白水平；(2)加入100 nM E2α、20 mM葡萄糖和5 mU葡萄糖氧化酶(glucose oxidase, GOX),在培养液中GOX可以分解G并产生H202,对细胞造成氧化损伤。24h后,进行DHE染色观察APP/PS1 N2a细胞中活性氧(ROS)含量的变化。
结果：不同浓度E2α(25nM,50nM,100nM,200nM)处理后,细胞外Ap蛋白水平明显减少,差异具有统计学意义(P<0.05)；而细胞内Ap蛋白水平无明显变化(P>0.05)；细胞BACE1的表达水平无明显变化(P>0.05)；E2α可以减少G/GOX所致APP/PS1 N2a细胞ROS的产生。
结论：E2α可以减少细胞外Ap的产生,其机制可能与其抗氧化损伤有关。
Objective To investigate the effects of 17a-estradiol (E2a) on the release ofβ-amyloid protein (Aβ) with the transgenic cell line APP/PS 1 N2a, and discuss the potential mechanism of such effects.
Methods APP/PS 1 N2a cell lines were pretreated with 10 nM E2a for 2 days, then changed the culture medium, and treated with different methods:(1) cell lines were treated with either E2a (25nM,50nM, 100nM,200nM) or vehicle, The protein level of Aβand BACE1 both intracellular and extracellular of APP/PS 1 N2a cells were detected by Western blot. (2) cell lines were treated with 100nM E2a,20 mM glucose and 5 mU glucose oxidase(GOX), followed by exposure to the glucose oxidase (G/Gox) to induce oxidative stress. After 24 hours, the change of the ROS contents in APP/PS 1 N2a cells was observed for DHE.
Results Pretreatment of APP/PS 1 N2a cells with different concentration (25nM,50nM, 100nM,200nM) E2a significantly reduced extracellular levels of AP(P<0.05). There was no change both in the intracellular levels of Aβand the levels of BACE1(P>0.05). E2a can reduce the ROS produced by APP/PS 1 N2a cells after G/Gox treatment.
Conclusion E2a can reduce the production of extracellular Aβ,its mechanism may be related to the oxidation damage.
引文
[1]Hyman BT New neuropathological criteria for Alzheimer disease. Arch Neurol,1998,55(09):1174-1176.
[2]郑君德.氧化应激与老年性痴呆发病的相关性研究.中国神经免疫学和神经病学杂志,2008,15(4)：301-304.
[3]Huagn Y, Xu YM, Zhang JW, Ren XH, Suo AQ. Effects of estrogen on P-Tau, ChAT and nerve growth factor protein expressions in the brain tissue of rats with Alzheimer's disease. Nan Fang Yi Ke Da Xue Xue Bao,2010,30(10):2408-10.
[4]Simon J, Nachtigall L, Ulrich LG, Eugster-Hausmann M, Gut R. Endometrial safety of ultra-low-dose estradiol vaginal tablets. Obstet Gynecol,2010,116(4):876-83.
[5]Gouras GK, Xu H, Gross RS, Greenfield JP, Hai B, Wang R, Greengard P. Testosterone reduces neuronal secretion of Alzheimers β-amyloid peptides. Proc Natl Acad Sci U S A.2000,97(3):1202-1205.
[6]沈颖华.烟碱型乙酰胆碱受体的激动与阿尔茨海默病的治疗.解放军药学学报,2008,24(2)：159-162.
[7]Zhang S, Huang Y, Zhu YC, Yao T. Estrogen stimulates release of secreted amyloid precursor protein from primary rat cortical neurons via protein kinase C pathway.Acta Pharmacol Sin.2005,26(2):171-176.
[8]黄艳红.卵巢切除及雌激素替代对大鼠脑单胺氧化酶和行为学的影响.中国行为医学科学,2004,10(2)：140-141.
[9]Gibbs RB. Effects of ageing and long-term hormone replacement on cholinergic neurones in the medial septum and nucleus basalis magnocellularis of ovariectomized rats. J Neuroendocrinol,2003,15(5):477-485.
[10]Green P.S, Gridley K.E. and Simpkins J.W. Nuclear estrogen receptor-independent neuroprotection by estratrienes:a novel interaction with glutathione. Neuroscience,1998,84(1):7-10.
[11]Behl C, Widmann M, Trapp T and Holsboer F.17-beta estradiol protects neurons from oxidative stress-induced cell death in vitro. Biochem Biophys Res Commun,1995,216 (2):473-482.
[12]Lu YP, Zeng M, Hu XY, Xu H, Swaab DF, Ravid R, Zhou JN. Estrogen receptor alpha-immunoreactive astrocytes are increased in the hippocampus in Alzheimer's disease. Exp Neurol,2003,183(2):482-488.
[13]McClean J, Nunez JL.17alpha-Estradiol is neuroprotective in male and female rats in a model of early brain injury. Exp Neurol,2008,210(1):41-50.
[14]Manthey D, Heck S, Engert S, Behl C. Estrogen induces a rapid secretion of amyloid beta precursor protein via the mitogen-activated protein kinase pathway. Eur J Biochem,2001,268(15):4285-4291.
[15]Singh M, Se'ta'lo'Jr G, Guan X, Warren M, Toran-Allerand CD. Estrogeninduced activation of mitogen-activated protein kinase in cerebral cortical explants:convergence of estrogen and neurotrophin signaling pathways. J Neurosci, 1999,19(4):1179-1188.
[16]Singh M. Ovarian hormones elicit phosphorylation of Akt and extracellular-signal regulated kinase in explants of the cerebral cortex. Endocrine,2001,14(3):407-415.
[17]Toran-Allerand CD, Guan X, MacLusky NJ, Horvath TL, Diano S, Singh M, Connolly Jr ES, Nethrapalli IS, Tinnikov AA. "ER-X":a novel, plasma membrane-associated, putative estrogen receptor that is regulated during development and after ischemic brain injury. J Neurosci,2002,22(19):8391-8401.
[18]Mosselman S, Polman J, Difekma R. Identification and characterization of a novel human estrogen receptor. FEBS Lett,1996,392(1):49-53.
[19]俞小瑞.β雌二醇的神经保护作用与磷脂酰肌醇-3-激酶的活性关系.第四军医大学学报,2005,26(9)：808-811.
[20]周开锋.17β-雌二醇对大鼠脊髓损伤后神经保护作用的研究.中国脊柱脊髓杂志,2007,17(8)：623-627.
[21]Numakawa Y, Matsumoto T, et al.17beta-estradiol protects cortical neurons against oxidative stress-induced cell death through reduction in the activity of mitogen-activated protein kinase and in the accumulation of intracellular calcium. Endocrinology,2007,148(2):627-637.
[22]Tung N, Miron A, Schnitt SJ, et al. Prevalence and predictors of loss of wild type BRCA1 in estrogen receptor positive and negative BRCA1-associated breast cancers. Breast Cancer Res.2010,12(6):95-96.
[23]C. Dominique Toran-Allerand, Alexander A. Tinnikov, et al.17α-Estradiol: A Brain-Active Estrogen. Endocrinology,2005,146(9):3843-3850.
[24]Simpkins JW, Rajakumar G, Zhang YQ, Simpkins CE, Greenwald D, Yu CJ, Bodor N, Day AL. Estrogens may reduce mortality and ischemic damagecaused by middle cerebral artery occlusion in the female rat. J Neurosurg,1997,87(5):724-730.
[25]Levin-Allerhand JA, Lominska CE, Wang J, Smith JD.17α-estradiol and 17β-estradiol treatments are effective in lowering cerebral amyloid-β levels in AβPPSWE transgenic mice. J Alzheimers Dis,2002,4(6):449-457.
[26]Gelinas S, Bureau G, Valsatro B, Massicotte G, Cicchetti F, Chiasson K, Gagne B, Blanchet J. α-And β-estradiol protect neuronal but not native PC 12 cells from paraquat-induced oxidative stress. Neurotox Matinoli MG,2004,6(2):141-148.
[27]Vassar R, Bennett BD, Babu, Khan S, et al. Beta-secretase cleavage of Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE. Science,1999,286 (5440):735-741.
[28]Mortimer JA, Snowdon DA, Markesbery WR. Head circumference, education and risk of dementia:findings from the Nun Study. J Clin Exp Neurepsychol,2003,25(5):671-679.
[29]Levy-Lahad E, Waseo W, Poorkaj P, et al. Candidate gene for the chromosome 1 familial Alzheimer's disease locus. Science,1995,269(5226):973-977.
[30]Willnow TE, Carlo AS, Rohe M, Schmidt V. SORLA/SORL1, a neuronal sorting receptor implicated in Alzheimer's disease. Rev Neurosci.2010,21(4):315-329.
[31]Lesn S, Koh MT, Kotilinek L, et al. A specific amyloid-beta protein assembly in the brain impairs memory. Nature,2006,440(7082):352-357.
[32]Tanzi RE, Bertram L. Twenty years of the Alzheimer's disease amyloid hypothesis:a genetic perspective [J]. Cell,2005,120(4):545-555.
[33]Boddapati S, Levites Y, Sierks MR. Inhibiting (3-secretase activity in Alzheimer's disease cell models with single chain antibodies specifically targeting APP. J Mol Biol,2010,171 (3):859-868.
[34]Hedskog L, Petersen CA, Svensson AI, Welander H, Tjernberg LO, Karlstrom H, Ankarcrona M. Gamma-secretase Complexes Containing Caspase-cleaved Presenilin-1 Increase Intracellular Aβ42/Aβ40 Ratio. J Cell Mol Med,2010, 10(34):1582-1584.
[35]Schettini G, Govoni S, Racchi M, Rodriguez G. Phosphorylation of APP-CTF-AICD domains and interaction with adaptor proteins:signal transduction and/or transcriptional role - relevance for Alzheimer pathology. J Neurochem,2010, 10(1):1471-1474.
[36]Asai M, Iwata N, Tomita T, Iwatsubo T, Ishiura S, Saido TC, Maruyama K. Efficient four-drug cocktail therapy targeting amyloid-β peptide for Alzheimer's disease. J Neurosci Res,2010,88(16):3588-3597.
[37]Skinner M. Oxidative stress:ATM bonds under stress. Nat Rev Mol Cell Biol,2010,11(12):818-819.
[38]Nunomura A, Perry G, Aliev G, et al. Oxidative damage is the earliest event in Alzheimer disease. J Neuropathol Exp Neurol,2001,60(8):759-767.
[39]Nunomura A, Perry G, Pappolla MA, et al. RNA oxidation is a prominent feature of vulnerable neurons in Alzheimer's disease. J Neurosci,1999,19(6):1959-1964.
[40]Yang X, Askarova S, Sheng W, Chen JK, Sun AY, Sun GY, Yao G, Lee JC. Low energy laser light (632.8 nm) suppresses amyloid-β peptide-induced oxidative and inflammatory responses in astrocytes. Neuroscience,2010,171(3):859-868.
[1]Michel BF, Luciani V, Geda YE, et al. In Alzheimer's disease, the clinical expression of behavioral and psychological signs and symptoms is early and specific of neuropathological stages. Encephale,2010,36(4):314-325.
[2]肖增平.老年痴呆的发病机制及治疗的研究进展.中国老年学杂志,2008,28(1)：155-157.
[3]Mortimer JA, Snowdon DA, Markesbery WR. Small head circumference is associated with less education in persons at risk for Alzheimer disease in later life. Alzheimer Dis Assoc Disord,2008,22(3):249-254.
[4]Bennet AM, Di Angelantonio E, Ye Z. Association of apolipoprotein E genotypes with lipid levels and coronary risk. JAMA,2007,298(11):1300-1311.
[5]Nawrot B. Targeting BACE with small inhibitory nucleic acids-a future for Alzheimer's disease therapy? Acta Biochim Pol,2004,51(2):431-444.
[6]Rovelet-Lecrux A, Hannequin D, Raux G, et al. APP locus duplication causes autosomal dominant early-onset Alzheimer disease with cerebral amyloid anglopethy. Nat Genet,2006,38(1):24-26.
[7]Steinhilb ML, Dias-Santagata D, Fulga TA Tau phosphorylation sites work in concert to promote neurotoxicity in vivo. Mol Bid Cell,2007,18(12):5060-5068.
[8]Butterfield DA, Reed T, Newman SF. Roles of amyloid beta-peptide-associated oxidative stress and brain protein modifications in the pathogenesis of Alzheimer's disease and mild cognitive impairment. Free Radic Biol Med,2007, 43(5):658-677.
[9]Yan SD, Stem DM. Mitechondrial dysfunction and Alzheimer's disease; role of amyloid-beta peptide alcohol dohydrogunase(ABAD). Int J Exp Pathol,2005, 86(3):161-171.
[10]Mocanu MM, Nissen A, Eckermann K, The potential for β-structure in the repeat domain of tau protein determines aggregation, synaptic decay, neuronal loss, and coassembly with endogenous Tau in inducible mouse models of tauopathy. J Neurosci,2008,28(3):737-748.
[11]Castellani RJ, Nunomura A, Lee HG, et al. Phosphorylated tau:Toxic, protective, or none of the above. J Alzheimer's Dis,2008,14(4):377-383.
[12]Clavaguera F, Bolmont T, Crowther RA, et al. Transmission and spreading of tauopathy in transgenic mouse brain. Nat Cell Biol,2009,11(7):909-913.
[13]Wang JZ, Grundke-Iqbal I, Iqbal K, et al. Kinases and phosphatases and tau sites involved in Alzheimer neurofibrillary degeneration. Eur J Neuresci,2007, 25(1):59-68.
[14]Jellinger KA Challenges in neuronal apoptosis. Curt Alzheimer Res,2006; 3(4):377-391.
[15]高清,吴波.细胞凋亡与细胞骨架的改变.医学研究生学报,2008,21(8)：877-880.
[16]Alonso AC, Li B. Grundke-Iqbal I Polymerization of hyperphosphorylated tau into filaments eliminates its inhibitory activity. Proc Natl Acad Sci USA,2006, 103(23):8864-8869.
[17]Ash AD, Aliev G., Siedlak SL Microtubule reduction in Alzheimer's disease and aging is independent of tau filament formation. Am J Pathol,2003,162(5):1623-1627.
[18]Lapointe NE, Morfini G., Pigino G. The amino terminus of tau inhibits kinesin-dependent axonal transport:Implications for filament toxicity. J Neurosci Res, 2009,87(2):440-451.
[19]Lane RM, Potkin SG, Enz A. Targeting acetylcholinesterase and butyrylcholinesterase in dementia. Int J Neuropsychopharmacol,2006,9(1):101-124.
[20]Yoo JH, Valdovinos MG., Williams DC, et al. Relevance of donepezil in enhancing learning and memory in special populations:a review of the literature. J Autism Dev Disord,2007,37(10):1883-1901.
[21]Elder GA, Gama Sosa MA, Gasperi R, et al. Presenilin transgenic mice as models of Alzheimer's disease. Brain Struct Funet,2010,214(2-3):127-143.
[22]Alvesda Costa C, Paitel E, Mattson MP, et al. Wild-type and mutated presenilins 2 trigger p53-dependent apoptosis and down-regulate presenilin 1 expression in HEK293 human cells and in murine neurons. Proc Natl Acad Sci USA, 2002,99(6):4043-4048.
[23]Van Leeuwen I, Lain S. Sirtuins and p53 Adv Cancer Res.2009,102:171-195.
[24]Yeung F. Hoberg JE.Ramsey CS Modulation of NF-kappaB-dependent transcription and cell survival by the Sirt 1 deacetylase. EMBO J,2004,23(12):2369-2380.
[25]Tang BL Alzheimer's disease：channeling APP to non-amyloidogenic processing. Biochem Biophys Res Commun,2005,331(2):375-378.
[26]Chakraborty M, Qiu SG. Vasudevan KM Par-4 drives trafficking and activation of Fas and Fasl toinduce prostate cancer cell apoptosis and tumor regression. Neuro pathol Exp Neuml,2001,61(19):7255-7263.
[27]Vetterkind S, Lee E, Sundberg E, et al. Par-4:a new activator of myosin phosphatase. Mol Biol Cell,2010,21(7):1214-1224.
[28]Wang G, Silva J, Krishnamurthy K, et al. Direct binding to ceramide activates protein kinase Czeta before the formation of a pro-apoptotic complex with PAR-4 in differentiating stem cells. J Biol Chem,2005,280(28):26415-26424.
[29]Wyss-Coray T, Yan F. Lin AH Prominent neurodegeneration and increased plaque formation in complement-inhibited Alzheimer's mice. Pmc Nail Acad Sei USA, 2002,99(16):10837-10842.
[30]Monte SM, Chen GJ, Rivera E. Neuronal thread protein regulation and interaction withmicrotubule-associated proteins in SH-Sy5y neuronal cells. Cell Mol Life Sci,2003,60(12):2679-1691.
[31]Greeve I, Hermans-Borgmeyer I, Brellinger C, et al. The human DIMINUTO/DWARF1 homolog seladin-1 confers resistance to Alzheimer's disease-associated neurodegeneration and oxidative stress. J Neurosci,2000,20(19):7345-7352.
[32]Butterfield DA, Reed T, Newman SF. Roles of amyloid beta-peptide-associated oxidative stress and brain protein modifications in the pathogenesis of Alzheimer's disease and mild cognitive impairment. Free Radic Biol Med,2007, 43(5):658-677.
[33]Chang WP, KoelschG, Wong S. In vivoinhibition of Abeta production by memapsin 2(beta-secretase) inhibitors. J Neurochem,2004,89(6):1409-1416.
[34]Chang WP, Koelsch G, Wong S, et al. In vivo inhibition of A-beta production by memapsin 2(beta—secretase) inhibitors. J Neurechem,2004, 89(6):1409-1416.
[35]Eteheberrigaray R, Tan M, Dewachter I, et al. Therapeutic effects of PKC activators in Alzheimer's disease transgenic mice. Proc Nad Acad Sci U S A,2004, 101(30):11141-11146.
[36]李玺,张欣,张智燕,袁海峰,权乾坤.人参皂苷Rgl对AD模型大鼠脑片P、Tau、PKA影响的实验研究.中国老年学杂志,2009,10(29)：2468-2470.
[37]刘艳玲,李奴英.乙酰胆碱酯酶抑制荆治疗老年痴呆症的研究进展.山西医药杂志,2008,1(37)64-66.
[38]方立,陈瑶,张奕华.抗阿尔茨海默病的胆碱酯酶抑制剂研究进展.药学进展,2009,7(33)289-295.
[39]Gray JA.Grigoryan G.Videy D Conditionally immortalized,multipotential and multifunctional neural stem cell lines as an approach to clinical transplantation. Cell Transplant,2000,9(2):153-168.
[40]Refolo LM, Pappolla MA, LaFrancois J, et al. A cholesterollowenng drug reduces beta-amyloid pathology in a transgenic mouse model of Alzheimer's disease. Neurobiol Dis,2001,8(5):890-899.
[41]Xiu J, Nordberg A, Shan KR, et al. Lovastatin stimulates up-regulation of alpha7 nicotinic receptors in cultured neurons without cholesterol dependency, a mechanism involving production of the alpha-form of secreted amyloid precursor protein. J Neurosci Res,2005,82(4):531-541.
[42]Roensch J, Crisby M, Nordberg A, et al. Effects of statins on alpha7 nicotinic receptor, cholinesterase and alpha-form of secreted amyloid precursor peptide in SH-SY5Y cells. Neurechem Int,2007,50(6):800-806.
[2]郑君德.氧化应激与老年性痴呆发病的相关性研究.中国神经免疫学和神经病学杂志,2008,15(4)：301-304.
[3]Huagn Y, Xu YM, Zhang JW, Ren XH, Suo AQ. Effects of estrogen on P-Tau, ChAT and nerve growth factor protein expressions in the brain tissue of rats with Alzheimer's disease. Nan Fang Yi Ke Da Xue Xue Bao,2010,30(10):2408-10.
[4]Simon J, Nachtigall L, Ulrich LG, Eugster-Hausmann M, Gut R. Endometrial safety of ultra-low-dose estradiol vaginal tablets. Obstet Gynecol,2010,116(4):876-83.
[5]Gouras GK, Xu H, Gross RS, Greenfield JP, Hai B, Wang R, Greengard P. Testosterone reduces neuronal secretion of Alzheimers β-amyloid peptides. Proc Natl Acad Sci U S A.2000,97(3):1202-1205.
[6]沈颖华.烟碱型乙酰胆碱受体的激动与阿尔茨海默病的治疗.解放军药学学报,2008,24(2)：159-162.
[7]Zhang S, Huang Y, Zhu YC, Yao T. Estrogen stimulates release of secreted amyloid precursor protein from primary rat cortical neurons via protein kinase C pathway.Acta Pharmacol Sin.2005,26(2):171-176.
[8]黄艳红.卵巢切除及雌激素替代对大鼠脑单胺氧化酶和行为学的影响.中国行为医学科学,2004,10(2)：140-141.
[9]Gibbs RB. Effects of ageing and long-term hormone replacement on cholinergic neurones in the medial septum and nucleus basalis magnocellularis of ovariectomized rats. J Neuroendocrinol,2003,15(5):477-485.
[10]Green P.S, Gridley K.E. and Simpkins J.W. Nuclear estrogen receptor-independent neuroprotection by estratrienes:a novel interaction with glutathione. Neuroscience,1998,84(1):7-10.
[11]Behl C, Widmann M, Trapp T and Holsboer F.17-beta estradiol protects neurons from oxidative stress-induced cell death in vitro. Biochem Biophys Res Commun,1995,216 (2):473-482.
[12]Lu YP, Zeng M, Hu XY, Xu H, Swaab DF, Ravid R, Zhou JN. Estrogen receptor alpha-immunoreactive astrocytes are increased in the hippocampus in Alzheimer's disease. Exp Neurol,2003,183(2):482-488.
[13]McClean J, Nunez JL.17alpha-Estradiol is neuroprotective in male and female rats in a model of early brain injury. Exp Neurol,2008,210(1):41-50.
[14]Manthey D, Heck S, Engert S, Behl C. Estrogen induces a rapid secretion of amyloid beta precursor protein via the mitogen-activated protein kinase pathway. Eur J Biochem,2001,268(15):4285-4291.
[15]Singh M, Se'ta'lo'Jr G, Guan X, Warren M, Toran-Allerand CD. Estrogeninduced activation of mitogen-activated protein kinase in cerebral cortical explants:convergence of estrogen and neurotrophin signaling pathways. J Neurosci, 1999,19(4):1179-1188.
[16]Singh M. Ovarian hormones elicit phosphorylation of Akt and extracellular-signal regulated kinase in explants of the cerebral cortex. Endocrine,2001,14(3):407-415.
[17]Toran-Allerand CD, Guan X, MacLusky NJ, Horvath TL, Diano S, Singh M, Connolly Jr ES, Nethrapalli IS, Tinnikov AA. "ER-X":a novel, plasma membrane-associated, putative estrogen receptor that is regulated during development and after ischemic brain injury. J Neurosci,2002,22(19):8391-8401.
[18]Mosselman S, Polman J, Difekma R. Identification and characterization of a novel human estrogen receptor. FEBS Lett,1996,392(1):49-53.
[19]俞小瑞.β雌二醇的神经保护作用与磷脂酰肌醇-3-激酶的活性关系.第四军医大学学报,2005,26(9)：808-811.
[20]周开锋.17β-雌二醇对大鼠脊髓损伤后神经保护作用的研究.中国脊柱脊髓杂志,2007,17(8)：623-627.
[21]Numakawa Y, Matsumoto T, et al.17beta-estradiol protects cortical neurons against oxidative stress-induced cell death through reduction in the activity of mitogen-activated protein kinase and in the accumulation of intracellular calcium. Endocrinology,2007,148(2):627-637.
[22]Tung N, Miron A, Schnitt SJ, et al. Prevalence and predictors of loss of wild type BRCA1 in estrogen receptor positive and negative BRCA1-associated breast cancers. Breast Cancer Res.2010,12(6):95-96.
[23]C. Dominique Toran-Allerand, Alexander A. Tinnikov, et al.17α-Estradiol: A Brain-Active Estrogen. Endocrinology,2005,146(9):3843-3850.
[24]Simpkins JW, Rajakumar G, Zhang YQ, Simpkins CE, Greenwald D, Yu CJ, Bodor N, Day AL. Estrogens may reduce mortality and ischemic damagecaused by middle cerebral artery occlusion in the female rat. J Neurosurg,1997,87(5):724-730.
[25]Levin-Allerhand JA, Lominska CE, Wang J, Smith JD.17α-estradiol and 17β-estradiol treatments are effective in lowering cerebral amyloid-β levels in AβPPSWE transgenic mice. J Alzheimers Dis,2002,4(6):449-457.
[26]Gelinas S, Bureau G, Valsatro B, Massicotte G, Cicchetti F, Chiasson K, Gagne B, Blanchet J. α-And β-estradiol protect neuronal but not native PC 12 cells from paraquat-induced oxidative stress. Neurotox Matinoli MG,2004,6(2):141-148.
[27]Vassar R, Bennett BD, Babu, Khan S, et al. Beta-secretase cleavage of Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE. Science,1999,286 (5440):735-741.
[28]Mortimer JA, Snowdon DA, Markesbery WR. Head circumference, education and risk of dementia:findings from the Nun Study. J Clin Exp Neurepsychol,2003,25(5):671-679.
[29]Levy-Lahad E, Waseo W, Poorkaj P, et al. Candidate gene for the chromosome 1 familial Alzheimer's disease locus. Science,1995,269(5226):973-977.
[30]Willnow TE, Carlo AS, Rohe M, Schmidt V. SORLA/SORL1, a neuronal sorting receptor implicated in Alzheimer's disease. Rev Neurosci.2010,21(4):315-329.
[31]Lesn S, Koh MT, Kotilinek L, et al. A specific amyloid-beta protein assembly in the brain impairs memory. Nature,2006,440(7082):352-357.
[32]Tanzi RE, Bertram L. Twenty years of the Alzheimer's disease amyloid hypothesis:a genetic perspective [J]. Cell,2005,120(4):545-555.
[33]Boddapati S, Levites Y, Sierks MR. Inhibiting (3-secretase activity in Alzheimer's disease cell models with single chain antibodies specifically targeting APP. J Mol Biol,2010,171 (3):859-868.
[34]Hedskog L, Petersen CA, Svensson AI, Welander H, Tjernberg LO, Karlstrom H, Ankarcrona M. Gamma-secretase Complexes Containing Caspase-cleaved Presenilin-1 Increase Intracellular Aβ42/Aβ40 Ratio. J Cell Mol Med,2010, 10(34):1582-1584.
[35]Schettini G, Govoni S, Racchi M, Rodriguez G. Phosphorylation of APP-CTF-AICD domains and interaction with adaptor proteins:signal transduction and/or transcriptional role - relevance for Alzheimer pathology. J Neurochem,2010, 10(1):1471-1474.
[36]Asai M, Iwata N, Tomita T, Iwatsubo T, Ishiura S, Saido TC, Maruyama K. Efficient four-drug cocktail therapy targeting amyloid-β peptide for Alzheimer's disease. J Neurosci Res,2010,88(16):3588-3597.
[37]Skinner M. Oxidative stress:ATM bonds under stress. Nat Rev Mol Cell Biol,2010,11(12):818-819.
[38]Nunomura A, Perry G, Aliev G, et al. Oxidative damage is the earliest event in Alzheimer disease. J Neuropathol Exp Neurol,2001,60(8):759-767.
[39]Nunomura A, Perry G, Pappolla MA, et al. RNA oxidation is a prominent feature of vulnerable neurons in Alzheimer's disease. J Neurosci,1999,19(6):1959-1964.
[40]Yang X, Askarova S, Sheng W, Chen JK, Sun AY, Sun GY, Yao G, Lee JC. Low energy laser light (632.8 nm) suppresses amyloid-β peptide-induced oxidative and inflammatory responses in astrocytes. Neuroscience,2010,171(3):859-868.
[1]Michel BF, Luciani V, Geda YE, et al. In Alzheimer's disease, the clinical expression of behavioral and psychological signs and symptoms is early and specific of neuropathological stages. Encephale,2010,36(4):314-325.
[2]肖增平.老年痴呆的发病机制及治疗的研究进展.中国老年学杂志,2008,28(1)：155-157.
[3]Mortimer JA, Snowdon DA, Markesbery WR. Small head circumference is associated with less education in persons at risk for Alzheimer disease in later life. Alzheimer Dis Assoc Disord,2008,22(3):249-254.
[4]Bennet AM, Di Angelantonio E, Ye Z. Association of apolipoprotein E genotypes with lipid levels and coronary risk. JAMA,2007,298(11):1300-1311.
[5]Nawrot B. Targeting BACE with small inhibitory nucleic acids-a future for Alzheimer's disease therapy? Acta Biochim Pol,2004,51(2):431-444.
[6]Rovelet-Lecrux A, Hannequin D, Raux G, et al. APP locus duplication causes autosomal dominant early-onset Alzheimer disease with cerebral amyloid anglopethy. Nat Genet,2006,38(1):24-26.
[7]Steinhilb ML, Dias-Santagata D, Fulga TA Tau phosphorylation sites work in concert to promote neurotoxicity in vivo. Mol Bid Cell,2007,18(12):5060-5068.
[8]Butterfield DA, Reed T, Newman SF. Roles of amyloid beta-peptide-associated oxidative stress and brain protein modifications in the pathogenesis of Alzheimer's disease and mild cognitive impairment. Free Radic Biol Med,2007, 43(5):658-677.
[9]Yan SD, Stem DM. Mitechondrial dysfunction and Alzheimer's disease; role of amyloid-beta peptide alcohol dohydrogunase(ABAD). Int J Exp Pathol,2005, 86(3):161-171.
[10]Mocanu MM, Nissen A, Eckermann K, The potential for β-structure in the repeat domain of tau protein determines aggregation, synaptic decay, neuronal loss, and coassembly with endogenous Tau in inducible mouse models of tauopathy. J Neurosci,2008,28(3):737-748.
[11]Castellani RJ, Nunomura A, Lee HG, et al. Phosphorylated tau:Toxic, protective, or none of the above. J Alzheimer's Dis,2008,14(4):377-383.
[12]Clavaguera F, Bolmont T, Crowther RA, et al. Transmission and spreading of tauopathy in transgenic mouse brain. Nat Cell Biol,2009,11(7):909-913.
[13]Wang JZ, Grundke-Iqbal I, Iqbal K, et al. Kinases and phosphatases and tau sites involved in Alzheimer neurofibrillary degeneration. Eur J Neuresci,2007, 25(1):59-68.
[14]Jellinger KA Challenges in neuronal apoptosis. Curt Alzheimer Res,2006; 3(4):377-391.
[15]高清,吴波.细胞凋亡与细胞骨架的改变.医学研究生学报,2008,21(8)：877-880.
[16]Alonso AC, Li B. Grundke-Iqbal I Polymerization of hyperphosphorylated tau into filaments eliminates its inhibitory activity. Proc Natl Acad Sci USA,2006, 103(23):8864-8869.
[17]Ash AD, Aliev G., Siedlak SL Microtubule reduction in Alzheimer's disease and aging is independent of tau filament formation. Am J Pathol,2003,162(5):1623-1627.
[18]Lapointe NE, Morfini G., Pigino G. The amino terminus of tau inhibits kinesin-dependent axonal transport:Implications for filament toxicity. J Neurosci Res, 2009,87(2):440-451.
[19]Lane RM, Potkin SG, Enz A. Targeting acetylcholinesterase and butyrylcholinesterase in dementia. Int J Neuropsychopharmacol,2006,9(1):101-124.
[20]Yoo JH, Valdovinos MG., Williams DC, et al. Relevance of donepezil in enhancing learning and memory in special populations:a review of the literature. J Autism Dev Disord,2007,37(10):1883-1901.
[21]Elder GA, Gama Sosa MA, Gasperi R, et al. Presenilin transgenic mice as models of Alzheimer's disease. Brain Struct Funet,2010,214(2-3):127-143.
[22]Alvesda Costa C, Paitel E, Mattson MP, et al. Wild-type and mutated presenilins 2 trigger p53-dependent apoptosis and down-regulate presenilin 1 expression in HEK293 human cells and in murine neurons. Proc Natl Acad Sci USA, 2002,99(6):4043-4048.
[23]Van Leeuwen I, Lain S. Sirtuins and p53 Adv Cancer Res.2009,102:171-195.
[24]Yeung F. Hoberg JE.Ramsey CS Modulation of NF-kappaB-dependent transcription and cell survival by the Sirt 1 deacetylase. EMBO J,2004,23(12):2369-2380.
[25]Tang BL Alzheimer's disease：channeling APP to non-amyloidogenic processing. Biochem Biophys Res Commun,2005,331(2):375-378.
[26]Chakraborty M, Qiu SG. Vasudevan KM Par-4 drives trafficking and activation of Fas and Fasl toinduce prostate cancer cell apoptosis and tumor regression. Neuro pathol Exp Neuml,2001,61(19):7255-7263.
[27]Vetterkind S, Lee E, Sundberg E, et al. Par-4:a new activator of myosin phosphatase. Mol Biol Cell,2010,21(7):1214-1224.
[28]Wang G, Silva J, Krishnamurthy K, et al. Direct binding to ceramide activates protein kinase Czeta before the formation of a pro-apoptotic complex with PAR-4 in differentiating stem cells. J Biol Chem,2005,280(28):26415-26424.
[29]Wyss-Coray T, Yan F. Lin AH Prominent neurodegeneration and increased plaque formation in complement-inhibited Alzheimer's mice. Pmc Nail Acad Sei USA, 2002,99(16):10837-10842.
[30]Monte SM, Chen GJ, Rivera E. Neuronal thread protein regulation and interaction withmicrotubule-associated proteins in SH-Sy5y neuronal cells. Cell Mol Life Sci,2003,60(12):2679-1691.
[31]Greeve I, Hermans-Borgmeyer I, Brellinger C, et al. The human DIMINUTO/DWARF1 homolog seladin-1 confers resistance to Alzheimer's disease-associated neurodegeneration and oxidative stress. J Neurosci,2000,20(19):7345-7352.
[32]Butterfield DA, Reed T, Newman SF. Roles of amyloid beta-peptide-associated oxidative stress and brain protein modifications in the pathogenesis of Alzheimer's disease and mild cognitive impairment. Free Radic Biol Med,2007, 43(5):658-677.
[33]Chang WP, KoelschG, Wong S. In vivoinhibition of Abeta production by memapsin 2(beta-secretase) inhibitors. J Neurochem,2004,89(6):1409-1416.
[34]Chang WP, Koelsch G, Wong S, et al. In vivo inhibition of A-beta production by memapsin 2(beta—secretase) inhibitors. J Neurechem,2004, 89(6):1409-1416.
[35]Eteheberrigaray R, Tan M, Dewachter I, et al. Therapeutic effects of PKC activators in Alzheimer's disease transgenic mice. Proc Nad Acad Sci U S A,2004, 101(30):11141-11146.
[36]李玺,张欣,张智燕,袁海峰,权乾坤.人参皂苷Rgl对AD模型大鼠脑片P、Tau、PKA影响的实验研究.中国老年学杂志,2009,10(29)：2468-2470.
[37]刘艳玲,李奴英.乙酰胆碱酯酶抑制荆治疗老年痴呆症的研究进展.山西医药杂志,2008,1(37)64-66.
[38]方立,陈瑶,张奕华.抗阿尔茨海默病的胆碱酯酶抑制剂研究进展.药学进展,2009,7(33)289-295.
[39]Gray JA.Grigoryan G.Videy D Conditionally immortalized,multipotential and multifunctional neural stem cell lines as an approach to clinical transplantation. Cell Transplant,2000,9(2):153-168.
[40]Refolo LM, Pappolla MA, LaFrancois J, et al. A cholesterollowenng drug reduces beta-amyloid pathology in a transgenic mouse model of Alzheimer's disease. Neurobiol Dis,2001,8(5):890-899.
[41]Xiu J, Nordberg A, Shan KR, et al. Lovastatin stimulates up-regulation of alpha7 nicotinic receptors in cultured neurons without cholesterol dependency, a mechanism involving production of the alpha-form of secreted amyloid precursor protein. J Neurosci Res,2005,82(4):531-541.
[42]Roensch J, Crisby M, Nordberg A, et al. Effects of statins on alpha7 nicotinic receptor, cholinesterase and alpha-form of secreted amyloid precursor peptide in SH-SY5Y cells. Neurechem Int,2007,50(6):800-806.