叶酸对神经干细胞Notch和MAPK信号通路相关基因及蛋白表达作用的研究
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摘要
目的研究叶酸对大鼠胎鼠、新生鼠神经干细胞(neural stem cells,NSCs)增殖分化中Notch和MAPK信号通路相关基因及蛋白表达作用的机制。
方法采用孕14-16d SD胎鼠及24h内同窝新生鼠全脑,进行NSCs原代培养。在正常培养条件下,分为4组:①对照组;②叶酸缺乏组,添加甲氨蝶呤为0.4μg/ml;③叶酸低剂量组,添加叶酸为4μg/ml:④叶酸高剂量组,添加叶酸为40μg/ml。在加入MAPK通路抑制剂U0126的条件下,分为4组:①对照组:②U0126组,只添加U0126为10μM;③U0126+低剂量叶酸组,添加U0126为10μM同时添加叶酸为4μg/ml:④U0126+高剂量叶酸组,添加U0126为10μM同时添加叶酸为40μg/ml。增殖第6d收集一半细胞,另一半加入含血清培养液继续培养6d,诱导细胞分化,第13d收集分化的细胞。叶酸、甲氨蝶呤等试剂均用培养液配制。首先按正常条件培养神经干细胞,每次换液加入叶酸时即加入10μM的U0126。采用克隆、酶切与测序等方法对RT-PCR产物进行鉴定:采用实时定量SYBR Creen I-PCR方法,测定NSCs MAPK信号通路相关基因(ERK1、ERK2)mRNA表达情况:采用Western Blot方法,正常培养条件下Notch信号通路相关蛋白(Notch1、HES5、Neurogenin1)的表达情况和加入MAPK通路抑制剂U0126条件下,该通路中相关蛋白(pERK1、pERK2)的表达情况。
结果1、普通RT-PCR扩增产物经克隆、酶切、测序显示,特异性好,扩增片段与预期结果相符。2、SYBR Creen I-PCR结果显示,在加入MAPK通路抑制剂U0126的条件下,叶酸对胎鼠增殖期NSCs ERK1/2 mRNA表达的影响各组之间无统计学差异(P＞0.05);叶酸对新生鼠增殖期NSCs ERK1/2 mRNA表达的影响,某些组间虽有统计学差异,但没有一定的规律,体现不出叶酸的作用。
3、Western Blot方法对正常培养条件下Notch信号通路和加入MAPK通路抑制剂U0126条件下该通路中相关蛋白进行了检测,结果显示:(1)Notch通路增殖期补充叶酸能够促进NSCs Notch1、HES5蛋白的表达(P＜0.05),分化晚期补充叶酸能够降低Neurogenin1蛋白的表达(P＜0.05),同时也能增加HES5蛋白的表达(P＜0.05)。(2)MAPK通路无论增殖期分化期,叶酸对胎鼠及新生鼠NSCs pERK1、pERK2蛋白表达的影响均无统计学差异(P＞0.05)。
结论1、在正常培养条件下,叶酸可能通过上调增殖期胎鼠、新生鼠NSCs Notch信号通路中Notch1、HES5蛋白的表达促进神经干细胞的增殖;而在NSCs的分化晚期,叶酸再一次上调HES5蛋白的表达,下调Neurogenin1蛋白的表达,促进NSCs向星形胶质细胞分化。
2、在加入MAPK通路抑制剂U0126条件下,叶酸对增殖期胎鼠、新生鼠NSCs ERK1/2mRNA的表达影响不明显;对增殖分化期胎鼠、新生鼠NSCspERK1、pERK2蛋白的表达影响也不明显。
3、在正常培养条件下,叶酸40μg/ml剂量组与4μg/ml剂量对胎鼠、新生鼠NSCs的增殖分化中Notch通路中相关蛋白的调节作用基本一致;同样,在加入MAPK通路抑制剂U0126条件下,这两个剂量组对胎鼠、新生鼠NSCs的增殖分化MAPK通路中相关蛋白表达的调节作用也基本一致。
因此,叶酸可能通过影响Notch和MAPK信号通路中相关基因及蛋白的表达从而促进NSCs的增殖、分化,这对神经系统损伤和神经退行性疾病的治疗开辟新的途径。
Objective To study the proliferation and differentiation effects of folate on neural stem cells(NSCs)via Notch and MAPK signal pathway in fetal rats and newborn rats in vitro.
Methods NSCs of fetal SD rats from E14d to E16d and 24h newborn rats were cultured.In the normal condition,NSCs were divided into four groups which were control group,folate-deficiency group supplemented by methotrexate for 0.4μg/ml in media,folate-low group supplemented by folate for 4μg/ml in media,folate-high group supplemented by folate for 40μg/ml in media.When cultured with the inhibitor U0126 of MAPK pathway,NSCs were also divided into four groups which were control group,U0126 group supplemented by U0126 for 10μmol/L in media, folate-low group supplemented by U0126 for 10μmol/L and folate for 4μg/ml in media,folate-high group supplemented by U0126 for 10μmol/L and folate for 40μg/ml in media.After proliferation of NSCs had lasted for 6d,half of the cells were collected and the left were cultured for another 6d by adding FBS in order to promote NSCs to differentiate.Identification of the production of RT-PCR was via clone, enzyme digestion,sequence.The levels of related mRNA expression in MAPK signal pathway(ERK1,ERK2)were detected by SYBR Green I-PCR method.The levels of some kinds of proteins expression in fetal and newborn rats were detected by western blot method.
Results 1.The result of general RT-PCR showed the production of PCR via clone,enzyme digest,sequence were specific and corresponded with anticipation.
2.The SYBR Green I-PCR showed as follows when NSCs were cultured with inhibitor U0126 of MAPK signal pathway:There was no significant difference among the four groups although some groups supplemented by folate in NSCs of fetal rats(P＞0.05);There was a significant difference among some groups in NSCs of newborn rats(P＜0.05),but the results could not show the effect of folate.
3.The western blot method showed when NSCs were cultured in normal condition and in the condition with inhibitor U0126 of MAPK signal pathway:(1) Notch pathway Folate could enhance the levels of Notch1 and HES5 proteins in proliferation in NSCs(P＜0.05);Folate could decline the levels of Neurogeninl and enhance the levels of HES5 in late differentiation period in NSCs(P＜0.05).(2) MAPK pathway No matter in proliferation or in differentiation,there was no significant difference among the four groups of the effect of folate on NSCs in fetal or newborn rats.
Conclusions 1.When cultured in normal condition,folate could enhance the levels of Notch1 and HES5 proteins of Notch pathway to promote proliferation of fetal and newborn NSCs in proliferation period;Folate could enhance the levels of HES5 protein and decline the levels of Neurogeninl protein to promote fetal and newborn NSCs to differentiate into astrocytes in late differentiation period.
2.When cultured with the inhibitor U0126 of MAPK pathway,the effect of folate on ERK1/2 mRNA in proliferation of NSCs in fetal and newborn rats was not significant;the effect of folate on the levels of pERK1/2 was not significant either, but there was a trend that folate could promote the levels of pERK1/2.
3.No matter in the normal cultured condition or the condition with inhibitor U0126,the effect of folate on the levels of ERK1/2 mRNA and some proteins in NSCs of fetal and newborn rats was not significant.
Therefore,folate could enhance the levels of some mRNA and proteins of Notch and MAPK signal pathway to promote proliferation and differentiation of NSCs in fetal and newborn NSCs,and this research will explore a new way of treat on CNS injury and neural degenerative disease.
方法采用孕14-16d SD胎鼠及24h内同窝新生鼠全脑,进行NSCs原代培养。在正常培养条件下,分为4组:①对照组;②叶酸缺乏组,添加甲氨蝶呤为0.4μg/ml;③叶酸低剂量组,添加叶酸为4μg/ml:④叶酸高剂量组,添加叶酸为40μg/ml。在加入MAPK通路抑制剂U0126的条件下,分为4组:①对照组:②U0126组,只添加U0126为10μM;③U0126+低剂量叶酸组,添加U0126为10μM同时添加叶酸为4μg/ml:④U0126+高剂量叶酸组,添加U0126为10μM同时添加叶酸为40μg/ml。增殖第6d收集一半细胞,另一半加入含血清培养液继续培养6d,诱导细胞分化,第13d收集分化的细胞。叶酸、甲氨蝶呤等试剂均用培养液配制。首先按正常条件培养神经干细胞,每次换液加入叶酸时即加入10μM的U0126。采用克隆、酶切与测序等方法对RT-PCR产物进行鉴定:采用实时定量SYBR Creen I-PCR方法,测定NSCs MAPK信号通路相关基因(ERK1、ERK2)mRNA表达情况:采用Western Blot方法,正常培养条件下Notch信号通路相关蛋白(Notch1、HES5、Neurogenin1)的表达情况和加入MAPK通路抑制剂U0126条件下,该通路中相关蛋白(pERK1、pERK2)的表达情况。
结果1、普通RT-PCR扩增产物经克隆、酶切、测序显示,特异性好,扩增片段与预期结果相符。2、SYBR Creen I-PCR结果显示,在加入MAPK通路抑制剂U0126的条件下,叶酸对胎鼠增殖期NSCs ERK1/2 mRNA表达的影响各组之间无统计学差异(P＞0.05);叶酸对新生鼠增殖期NSCs ERK1/2 mRNA表达的影响,某些组间虽有统计学差异,但没有一定的规律,体现不出叶酸的作用。
3、Western Blot方法对正常培养条件下Notch信号通路和加入MAPK通路抑制剂U0126条件下该通路中相关蛋白进行了检测,结果显示:(1)Notch通路增殖期补充叶酸能够促进NSCs Notch1、HES5蛋白的表达(P＜0.05),分化晚期补充叶酸能够降低Neurogenin1蛋白的表达(P＜0.05),同时也能增加HES5蛋白的表达(P＜0.05)。(2)MAPK通路无论增殖期分化期,叶酸对胎鼠及新生鼠NSCs pERK1、pERK2蛋白表达的影响均无统计学差异(P＞0.05)。
结论1、在正常培养条件下,叶酸可能通过上调增殖期胎鼠、新生鼠NSCs Notch信号通路中Notch1、HES5蛋白的表达促进神经干细胞的增殖;而在NSCs的分化晚期,叶酸再一次上调HES5蛋白的表达,下调Neurogenin1蛋白的表达,促进NSCs向星形胶质细胞分化。
2、在加入MAPK通路抑制剂U0126条件下,叶酸对增殖期胎鼠、新生鼠NSCs ERK1/2mRNA的表达影响不明显;对增殖分化期胎鼠、新生鼠NSCspERK1、pERK2蛋白的表达影响也不明显。
3、在正常培养条件下,叶酸40μg/ml剂量组与4μg/ml剂量对胎鼠、新生鼠NSCs的增殖分化中Notch通路中相关蛋白的调节作用基本一致;同样,在加入MAPK通路抑制剂U0126条件下,这两个剂量组对胎鼠、新生鼠NSCs的增殖分化MAPK通路中相关蛋白表达的调节作用也基本一致。
因此,叶酸可能通过影响Notch和MAPK信号通路中相关基因及蛋白的表达从而促进NSCs的增殖、分化,这对神经系统损伤和神经退行性疾病的治疗开辟新的途径。
Objective To study the proliferation and differentiation effects of folate on neural stem cells(NSCs)via Notch and MAPK signal pathway in fetal rats and newborn rats in vitro.
Methods NSCs of fetal SD rats from E14d to E16d and 24h newborn rats were cultured.In the normal condition,NSCs were divided into four groups which were control group,folate-deficiency group supplemented by methotrexate for 0.4μg/ml in media,folate-low group supplemented by folate for 4μg/ml in media,folate-high group supplemented by folate for 40μg/ml in media.When cultured with the inhibitor U0126 of MAPK pathway,NSCs were also divided into four groups which were control group,U0126 group supplemented by U0126 for 10μmol/L in media, folate-low group supplemented by U0126 for 10μmol/L and folate for 4μg/ml in media,folate-high group supplemented by U0126 for 10μmol/L and folate for 40μg/ml in media.After proliferation of NSCs had lasted for 6d,half of the cells were collected and the left were cultured for another 6d by adding FBS in order to promote NSCs to differentiate.Identification of the production of RT-PCR was via clone, enzyme digestion,sequence.The levels of related mRNA expression in MAPK signal pathway(ERK1,ERK2)were detected by SYBR Green I-PCR method.The levels of some kinds of proteins expression in fetal and newborn rats were detected by western blot method.
Results 1.The result of general RT-PCR showed the production of PCR via clone,enzyme digest,sequence were specific and corresponded with anticipation.
2.The SYBR Green I-PCR showed as follows when NSCs were cultured with inhibitor U0126 of MAPK signal pathway:There was no significant difference among the four groups although some groups supplemented by folate in NSCs of fetal rats(P＞0.05);There was a significant difference among some groups in NSCs of newborn rats(P＜0.05),but the results could not show the effect of folate.
3.The western blot method showed when NSCs were cultured in normal condition and in the condition with inhibitor U0126 of MAPK signal pathway:(1) Notch pathway Folate could enhance the levels of Notch1 and HES5 proteins in proliferation in NSCs(P＜0.05);Folate could decline the levels of Neurogeninl and enhance the levels of HES5 in late differentiation period in NSCs(P＜0.05).(2) MAPK pathway No matter in proliferation or in differentiation,there was no significant difference among the four groups of the effect of folate on NSCs in fetal or newborn rats.
Conclusions 1.When cultured in normal condition,folate could enhance the levels of Notch1 and HES5 proteins of Notch pathway to promote proliferation of fetal and newborn NSCs in proliferation period;Folate could enhance the levels of HES5 protein and decline the levels of Neurogeninl protein to promote fetal and newborn NSCs to differentiate into astrocytes in late differentiation period.
2.When cultured with the inhibitor U0126 of MAPK pathway,the effect of folate on ERK1/2 mRNA in proliferation of NSCs in fetal and newborn rats was not significant;the effect of folate on the levels of pERK1/2 was not significant either, but there was a trend that folate could promote the levels of pERK1/2.
3.No matter in the normal cultured condition or the condition with inhibitor U0126,the effect of folate on the levels of ERK1/2 mRNA and some proteins in NSCs of fetal and newborn rats was not significant.
Therefore,folate could enhance the levels of some mRNA and proteins of Notch and MAPK signal pathway to promote proliferation and differentiation of NSCs in fetal and newborn NSCs,and this research will explore a new way of treat on CNS injury and neural degenerative disease.
引文
1.Gage FH.Mammalian Neural StemCells[J].Science,2000,287(5457):1433-1438.
2.裴丽君,李竹.还原叶酸载体基因(RFCI)与神经管和颅面畸形病因学关系的研究进展[J].遗传,2004,26(2):239-243.
3.Craciunescu CN,Brown EC,Mar MH,et al.Folic Acid Deficiency During Late Gestation Decreases Progenitor Cell Proliferation and Increases Apoptosis in Fetal Mouse Brain[J].Nutr,2004,134(1):162-166.
4.王树华,单培彦.叶酸、Vitamin B12与血管性痴呆[J].卒中与神经疾病.2005,12(5):311-313.
5.Oishi K,Kamakura S,Isazawa Y,et al.Notch promotes survival of neural precursor cells via mechanisms distinct from those regulating neurogenesis[J].Development biology,2004,276(1):172-184.
6.Jin Y,Yan EZ,Fan Y,et al.Sodium ferulate prevents amyloid-beta-induccd neurotoxicity through suppression of p38 MAPK and upregulation of ERK-1/2and Akt/proteinkinase B in rat hippocampus[J].Acta Pharmacologica Sinica,2005,26(8):943-951.
7.Galabova-Kovacs G,Matzen D,Piazzolla D,et al.Essential role of B-Raf in ERK activation during extraembryonic development[J].PNAS,2006,103(5):1325-1330.
8.Reynolds BA,Weiss S.Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system[J].Science,1992,225(5052):1707-1710.
9.Eriksson PS,Perfilieva E,Bjork-Eriksson T,et al.Neurogenesis in the adult human hippocampus[J].Nat Med,1998,4(11):1207-1313.
10.Thomson JA,Itskovitz-Eldor J,Shapiro SS,et al.Embryonic stem cell lines derived from human blastocysts[J].Science,1998,282(5391):1145-1147.
11.Toda H,Takahashi J,Iwakami N,et al.Grafting neural stem cells improved the impaired spatial recognition in ischemic rats[J].Neurosci Lett,2001,316:9-12.
12.陈丽.神经干细胞在神经内科疾病中的应用[J].中国城乡企业卫生,2004,6.28.
13. Grandbarbe L, Bouissac J, Rand M, et al. Delta-Notch signaling controls the generation of neurons/glia from neural stem cells in a stepwise process[J]. Development, 2003,130 (7):1391-1402.
14. Lutolf S, Radtke F, Aguet M, et al. Notchl is required for neuronal and glial differentiation in the cerebellum[J]. Development, 2002,129(2):373-385.
15. Tanigaki K, Nogaki F, Takahashi J, et al. Notchl and Notch3 instructively restrict bFGF-responsive multipotent neural progenitor cells to an astroglial fate [J]. Neuron, 2001, 29(1):45-55.
16. Daniel Rueda, Beatriz Navarro, Alberto Martinez-Serrano, et al. The Endocannabinoid Anandamide Inhibits Neuronal Progenitor Cell Differentiation through Attenuation of the Rap1/B-RaffERK Pathway [J]. Biol Chem, 2002, 277(48):46645-466650.
17. Uyttendaele H, Marazzi G, Wu G, et al. Notch4/int-3, a mammary proto-oncogene, is an endothelial cell-specific mammalian Notch gene[J]. Development, 1996,122(7):2251-2259.
18. Artavanis-Tsakonas S, Rand MD, Lake RJ. Notch signaling: cell fate control and signal integration in development[J]. Science, 1999, 284(5415):770-776 .
19. Iso T, Kedes L and Hamamori Y. HES and HERP families: multiple effectors of the Notch signaling pathway [J]. Cell Physiol, 2003,194(3):237-255.
20. Gaiano N, Fishell G The role of Notch in promoting glial and neural stem cell fates [J]. Annu Rev Neurosci, 2002, 25:471-490.
21. Gaiano N, Nye JS, Fishell G. Radial glial identity is promoted by Notchl signaling in the murine forebrain [J]. Neuron, 2000, 26(2):395-404.
22. Nam Y, Weng AP, Aster JC, et al. Structural requirements for assembly of the CSL, intracelular Notchl.Mastermind like transcriptional activation complex [J]. Biol Chem, 2003, 278(23):21232-21239.
23. Chambers CB, Peng Y, Nguyen H, et al. Spatiotemporal selectivity of response to Notchl signals in mammalian forebrain precursors [J]. Development, 2001, 128(5):689-702.
24. Ohtsuka T. Ishibashi M. Gradwohl G. et al. Hesl and Hes5 as notch effectors in mammalian neuronal differentiation[J]. EMBO J. 1999, 18:2196-2207.
25. Appel B, Eisen JS. Regulation of neuronal specification in the zebrafish spinal cord by Delta function[J]. Development, 1998.125:371-380.
26. Berezovska O, McLe an P, Knowles R, et al. Notchl inhibits neurite outgrowth in postmitotic primary neurons[J]. Neuroscience, 1999, 93:433-439.
27. Hitoshi S, Alexson T, Tropepe V. Notch pathway molecules are essential for the maintenance, but not the generation of mammalian neural stem cells[J]. Genes Dev, 2002, 16:846-858.
28. Toshiyuki Ohtsuka, Mkoto Ishibashi, Gerald Gradwohl et al. Hesl and Hes5 as Notch effectors in mammalian neuronal differentiation[J]. EMBO, 1999, 18(8):2196-2207.
29. Stephen T. Crews. Control of cell lineage-specific development and transcription by bHLH-PAS proteins[J]. Cold Spring Harbor Laboratory Press, 2008, 4:607-620.
30. Toshiyuki Ohtsuka, Masami Sakamoto, Francois Guillemot, et al. Roles of the Basic Helix-Loop-Helix Genes Hesl and Hes5 in Expansion of Neural Stem Cells of the Developing Brain[J]. Biological Chemistry, 2001,276(32):30467— 30474.
31. Farah MH, Olson JM, Sucic HB, et al. Generation of neutons by transient expression of neural bHLH proteins in mammaliancels[J]. Development, 2000, 127: 693-702.
32. Sun Y, NadM-Vicens M, Misono S, et al. Neurogenin promotes neurogenesis and inhibits glial differentiation by independent mechanisms[J]. Cell, 2001, 104: 365-376.
33. Nieto M, Schuurmans C, Britz O, et al. Neural bHLH genes control the neuronal versus glial fate decision in cortical progenitors [J]. Neuron, 2001, 29: 401-413.
34. Bylund M, Andersson E, Novitch BG, et al. Vertebrate neurogenesis is counteracted by Soxl-3 activity[J]. Nat Neurosci, 2003, 6:1162-1168.
35. Llado J, Haenggeli C, Maragakis NJ, et al. Neural stem cells protect against glutamate-induced excitotoxicity and promote survival of injured motor neurons through the secretion of neurotrophic factors[J]. Mol Cell Neurosci, 2004, 27(3): 322-331.
36.朱春梅,王天有.营养性巨幼红细胞性贫血的防治[J].中国实用乡村医生杂志,2004,11(4):9-10.
37.Tamura T,Picciano MF.Folate and human reproduction[J].Am J Clin Nutr,2006,83(5):993-1016.
38.Snowdon DA,Greiner LH,Mortimer JA,et al.Brain infarction and the clinical expression of Alzheimer disease.The Nun Study[J].JAMA,1997,277(10):813-817.
39.Zoccolella S,Lamberti P,Iliceto G,et al.Plasma homocysteine levels in L-dopa-treated Parkinson's disease patients with cognitive dysfunctions[J].Clin Chem Lab Med,2005,43(10):1107-1110.
40.Ravaglia G,Forti P,Maioli F,et al.Homocysteine and folate as risk factors for dementia and Alzheimer disease[J].Am J Clin Nutr,2005,82(3):636-643.
41.Mattson MP and Haberman F.Folate and Homocysteine Metabolism:Therapeutic Targets in Cardiovascular and Neurodegenerative Disorders[J].Curr Medl Chem,2003,10(19):1923-1929.
42.黄国伟,刘欢.叶酸、维生素B_6、B_(12)对脑缺血大鼠学习记忆的影响[J].中华预防医学杂志,2007,41(3):212-214.
43.Bottiglieri T,Laundy M,Crellin R,et al.Homocysteine,folate,methylation and monomunine metabolism in depression[J].J Neurol Neurosurg Psychiatry,2000,69:228-232.
44.Alpert JE,Mischoulon D,Nierenherg AA,et al.Nutrition and depression:focus on folate[J].Nutrition,2000,16:544-546.
45.Fava M,BoresJS,Alpert JE,et al.Folate,vitamin B_(12)and homocysteine in major depressive disorder[J].Am J Psychiatry,1997,154:426-428.
46.Kruman Ⅱ,Mouton PR,Emokpae R Jr,et al.Folate deficiency inhibits proliferation of adult hippocampal progenitors[J].Neuroreport,2005,16(10):1055-1059.
47.Shea TB,Rogers E.Folate quenches oxidative damage in brains of apolipoprotein E-deficient mice:augmentation by vitamin E[J].Brain Res Mol Brain Res,2002,108(1-2):1-6.
48.Ho PI,Collins SC,Dhitavat S,et al.Homocysteine potentiates beta-amyloid neurotoxicity,role of oxidative stress[J].Neurochem,2001,78(2):249-253.
49.Kruman Ⅱ,Kumaravel TS,Lohani A,et al.Folic acid deficiency and homocysteine impair DNA repair in hippocampal neurons and sensitize them to amyloid toxicity in experimental models of Alzheimer's disease[J].Neurosci,2002,22(5):1752-1762.
50.Keating E,Lemos C,Azevedo I,et al.Comparison of folic acid uptake characteristics by human placental choriocarcinoma cells at acidic and physiological Ph[J].Can J Physiol Pharmacol,2006,84(2):247-55.
51.张琦,郑志竑.Notch信号通路与神经干细胞的增殖分化[J].国外医学遗传学分册,2004,27(2):68-71.
52.Wada T,Penninger J M.Mitogen-activated protein kinases in apoptosis regulation[J].Oncogene,2004,23(16):2838-2849.
53.Seger R,Krebs E G.The MAPK signaling cascade[J].FASEB J,1995,9(9):726-735.
54.Ping Hu,Zhang Han,Anthony D.et al.Critical Role of Endogenous Akt/IAPs and MEK1/ERK Pathways in Counteracting Endoplasmic Reticulum Stress-induced Cell Death[J].Biological chemistry,2004,279(47):49420-49429.
55.Campos LS,Leone DP,Relvas JB,et al.Betal integrins activate a MAPK signalling pathway in neural stem cells that contributes to their maintenance[J].Development,2004,131(14):3433-3444.
56.Eunhae Kim,Amy L.Clark,et al.Belcheva.μ-and κ-Opioids Induce the Differentiation of Embryonic Stem Cells to Neural Progenitors[J].Biological Chemistry,2006,281(44):33749-33760.
57.端木德强,李中海,王敬泽.MAPK信号转导通路研究进展[J].细胞生物学杂志,2002,24(3):151-155.
58.姜勇,龚小卫.MAPK信号转导通路对炎症反应的调控[J].生理学报,2002,52(4):267-271.
59.牟金叶,陈晓光.促分裂原激活的蛋白激酶(MAPK)信号转导通路的研究进展[J].生命科学,2002,14(4):208-21.
60.张璐,姜勇,张琳.丝裂原活化蛋白激酶信号转导通路研究进展[J].国外医 学(生理、病理科学与临床分册),1999,19(2):84-87.
61.Adachi T,Kar S,Wang M,et al.Transient and sustained ERK phosphorylation and nuclear translocation in growth control[J].J Cell Physiol,2002,192(2):151-159.
62.孙小妹,毛萌,周晖等.脑源性神经营养因子对缺氧胚脑皮质神经元发挥保护作用的细胞内信号传导机制[J].华西医学,2006,21(3):454-457.
63.王彪,邢伟,许金华等.铝对大鼠海马ERK蛋白及mRNA表达影响[J].中国公共卫生,2006,22(12):1464-1466.
64.Vartiainen N,Goldsteins G,Keksa-Goldsteine V,et al.Aspirin Inhibits p44/42Mitogen-Activated Protein Kinase and Is Protective Against Hypoxia/Reoxygenation Neuronal Damage[J].Stroke,2003,34(3):752-757.
65.Madan P,Calder MD,Watson AJ.Mitogen-activated protein kinase(MAPK)blockade of bovine preimplantation embryogenesis requires inhibition of both p38 and extracellular signal-regulated kinase(ERK)pathways[J].Reproduction,2005,130(1):41-51.
66.Song HJ,Stevens CF,Gage FH.Neural stem cells from adult hippocampus develop essential properties of functional CNS neurons[J].Nat Neurosci,2002,5(5):438-445.
67.York RD,Yao H,Dillon T,et al.Rapl mediates sustained MAP kinase activation induced by nerve growth factor[J].Nature,1998,392(6676):622-626.
68.Zhifeng Xiao,Yaxian Kong,Shufa Yang,et al.Upregulation of Flk-1 by bFGF via the ERK pathway is essential for VEGF-mediated promotion of neural stem cell proliferation[J].Cell Research,2007,17(1):73-79.
69.Miguel A.Lopez-Toledano,Carolina Redondo,et al.Tyrosine Hydroxylase Induction by Basic Fibroblast Growth Factor and Cyclic AMP Analogs in Striatal Neural Stem Cells:Role of ERK1/ERK2 Mitogen-activated Protein Kinase and Protein Kinase C[J].Histochemistry & Cytochemistry,2004,52(9):1177-1189.
70.Borsello T,Clarke PG,Hirt L,et al.A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia[J].Nat Med,2003,9(9):1180-1186.
71.Ferrer I,Friguls B,Dalfo E,et al.Early modifications in the expression of mitogen-activated protein kinase(MAPK/ERK), stress-activated kinases SAPK/JNK and p38, and their phosphorylated substrates following focal cerebral ischemia[J]. Acta Neuropathol(Bed), 2003,105(5):425-437.
72. HuB, LiuC, Bramlett H, et al. Changes in trkB-ERK1/2-Creb/Elk-l pathways in hippocampal mossy fiber organization after traumatic brain injury[J]. J Cereb Blood Flow Metab, 2004,24(8):934-943.
73. Swatton JE, Sellers LA, Faun RL, 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.
74. Raghupathi R, Muir J K, Fulp C T, et al. Acute activation of mitogen-activated protein kinases following traumatic brain injury in the rat: implications for posttraumatic cell death[J]. Exp Neurol, 2003,183(2):438-448.
75. Raghupathi R, Graham DI, Mclntosh TK. Apoptosis after traumatic brain injury[J]. J Neurotrauma, 2000,17(10):927-938.
76. Marie-Therese Stockhausen, Jonas Sjolund and Hakan Axelson. Regulation of the Notch target gene Hes-1 by TGFa induced Ras/MAPK signaling in human neuroblastoma cells [J]. Experimental Cell Research, 2005,310 (1): 218-228.
1. Artavanis-Tsakonas S,Rand MD, Lake RJ. Notch signaling: cell fate control and signal integration in development[J]. Science, 1999,284:770-776.
2. Allman D, Aster JC, Pear W S. Notch signaling in hematopoiesis and early lymphocyte development[J]. Immunol Rev, 2002,187:75-86.
3. Arias A, Zecchini V, Brennan K. CSL-independent Notch signalling:a checkpoint in cell fate decisions during development[J]? Curr Opin Genet Dev. 2002,12:524-533.
4. Kojika S, Grifin JD. Notch receptors and hematopoiesis[J]. Exp Hematol, 2001, 29:1041-1052.
5. Lai EC. Notch cleavage:Nieastrin helps Presenilin make the final cut[J]. Curr Biol. 2002,12: R200-R202.
6. Milner LA, Bigas A. Notch as a mediator of cell fate determination in hema-Topoiesis: evidence and speculation[J]. Blood, 1999, 93: 2431-2448.
7. Bray S, Furriols M. Notch pathway: making sense of suppressor of hairless[J]. Curt Biol, 2001, 11 :R217-R221.
8. Okajima T, Irvine KD. Regulation of notch signaling by olinked fucose[J], Cell. 2002,111:893-904.
9. Shi S, Stanley P. Protein O-fueosyhransferasel is an essential component of Notch signaling pathways[J]. Proc Natl Acad Sci USA. 2003, 100: 5234-5239.
10. Lin SE, Oyama T, Nagase T, et al. Identification of new human mastermind proteins defines a family that consists of positive regulators for Notch signaling[J]. J Biol Chem, 2002,277: 50612-50620.
11. Jafar-Nejad H, Norga K, Bellen H. Numb:"Adapting" Notch for Endocytosis[J]. Dev Cell. 2002,3:155-156.
12. Yamamoto N, Yamamoto S, Inagaki F, et al .Role of Deltex-1 as a transcriptional regu lator downstream of the Notch receptor[J]. J Biol Chem, 2001, 276:45031-45040.
13. Matsuno K, Diederich RJ, Go MJ, et al. Dehex acts as a positive regulator of Notch signaling through interactions with the Notch ankyrin repeats[J]. Development,1995,121:2633-2644.
14.Matsuno K,Ito M,Hori K,et al.Involvement of a praline-rich motif and RING-H2 finger of Dehex in the regulation of Notch signaling[J].Development,2002,129:1049-1059.
15.Sestan N,Artavanis-Tsakonas S,Rakic P.Contact-de-pendent inhibition of cortical neurite growth mediated by Notch signaling[J].Science,1999,286:741-746.
16.Izon DJ,Aster JC,He Y,et al.Dehexl redirects lymphoid progenitors to the B cell lineage by antagonizing Notchl[J].Immunity,2002,16:231-243.
17.Ravaglia G,Forti P,Maioli F,Martelli M,Servadei L,Brunetti N,Porcellini E,Licastro F.Homocysteine and folate as risk factors for dementia and Alzheimer disease[J].Am J Clin Nutr,2005,82(3):636-643.
18.Mattson MP and Haberman F.Folate and Homocysteine Metabolism:Therapeutic Targets in Cardiovascular and Neurodegenerative Disorders[J].Curr Medl Chem,2003,10(19):1923-1929.
19.黄国伟,刘欢.叶酸、维生素B_6、B_(12)对脑缺血大鼠学习记忆的影响[J].中华预防医学杂志,2007,41(3):212-214.
20.Hitoshi S,Alexson T,Tropepe V,Donoviel D,Elia AJ,Nye JS,Conlon RA,Mak TW,Bernstein A,van der Kooy D.Neural stem cell lineages are regionally specified,but not committed,within distinct compartments of the developing brain[J].Development,2002,129(1):233-244.
21.王海燕,徐如祥,姜晓丹.Notch信号转导通路对神经干细胞的调控[J].生命的化学,2003,23(3):167-169.
22.Tsai RY,McKay RD.Cell contact regulates fate choice by cortical stem cells[J].J Neurosci,2000,20(10):3725-3735.
23.Alexson TO,Hitoshi S,Coles BL,et al.Notch signaling is required to maintain all neural stem cell populations irrespective of spatial or temporal niche[J].Dev Neurosci,2006,28(1-2):34-48.
24.张琦,郑志竑.Notch信号通路与神经干细胞的增殖分化[J].国外医学遗传学分册,2004,27(2):68-71.
25.刘素芳.Notch信号与神经干细胞分化[J].河南医学研究,2004,13(4),366-368.
26. Johnson JE, Biren SJ, Anderson DJ. Two rat homologues of drosophila achaete-scute specifically expressed in neuronal precursors[J]. Nature, 1990, 346(6287):858-861.
27. Douziech N, Calvo E, Laine J, et al. Activation of MAP kinases in growth[J]. Cell Signal, 1999,11:591-602.
28. Rampoldi, L, Zimbello R, Bortoluzzi S, et al. Chromosomal localization of four MAPK signaling cascade genes: MEK1, MEK3, MEK4 and MEKK5[J]. 1997, Cytogenetics and Cell Genetics[J], 78:301-303.
29. VitNe, G. Fagligt program - parallelle sesjoner[J].1999, Journal of Applied Microbiology, 87:288-293.
30. Canagarajah, B. J, et al. Activation mechanism of the MAP kinase ERK2 by dual phosphorylation[J]. 1997, Cell, 90:859-869.
31. Andrei, V. Khokhlatchev, Bertram Canagarajah, Julie Wilsbacher, et al. Activation Mechanism of the MAP Kinase ERK2 by Dual Phosphorylation[J]. 1998, Cell, 93:605-615.
32. Maria Deak, Andrew D, Clifton, John M, Lucocq et al. Mitogen- and stress-activated protein kinase-1 (MSK1) is directly activated by MAPK and SAPK2/p38, and may mediate activation of CREB[J]. 1998, EfBO, 17(15):4426-4441.
33. Reiko Sugiura, Takashi Toda, Susheela Dhut, et al. The MAPK kinase Pekl acts as a phosphorylation-dependent molecular switch[J]. 1999, Letters to nature, 399:479- 482.
34. SHUICHAN Xu, David Robbins, Jeff Frost, et al. MEKK1 phosphorylates MEK1 and MEK2 but does not cause activation of mitogen-activated protein kinase[J]. 1995, PNAS, 92: 6808-6812.
35. Borsello T, Clarke PG, Hirt L, et al. A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia[J]. Nat Med. 2003, 9(9):1180-1186.
36. Ferrer I, Friguls B, Dalfo E, et al. Early modifications in the expression of mitogen-activated protein kinase(MAPK/ERK), stress-activated kinases SAPK/JNK and p38, and their phosphorylated substrates following focal cerebral ischemia[J].Acta Neuropathol(Bed),2003,105(5):425-437.
37.HuB,LiuC,Bramlett H,et al.Changes in trkB-ERK1/2-cREB/Elk-1 pathways in hippocampal mossy fiber organization after traumatic brain injury[J].J Cereb Blood Flow Metab,2004,24(8):934-943.
38.Swatton JE,Sellers LA,Faun RL,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.
39.Raghupathi R,Muir J K,Fulp CT,et al.Acute activation of mitogen-activated protein kinases following traumatic brain injury in the rat:implications for posttraumatic cell death[J].Exp Neurol,2003,183(2):438-448.
40.Raghupathi R,Graham DI,Mclntosh TK.Apoptosis after traumatic brain injury[J].J Neurotrauma,2000,17(10):927-938.
41.孙小妹,毛萌,周晖等.脑源性神经营养因子对缺氧胚脑皮质神经元发挥保护作用的细胞内信号传导机制[J].华西医学,2006,21(3):454-457.
42.顾平,王彦永,马芹颖等.ERK1/2信号转导通路对骨髓基质细胞条件培养液诱导中脑神经干细胞分化作用的影响[J].2007,28(6):589-592.
43.Almeida RD,Manadas BJ,Melo CV,et al.Neuroprotection by BDNF against glutamate-induced apoptotic cell death is mediated by ERK and PI3-kinase pathways[J].Cell Death Difer,2005,12(10):1329-43.
44.Sakamoto T,Kawasoe Y,Shen JS,et al.Adenoviral gene transfer of GDNF,BDNF an d TGF beta 2,but not CNTF,cardiotrophin-1 or IGF1,protects injured adult motoneurons after facial nerve avulsion[,J].J Neuroeci Res,2003,72(1):54-64.
45.蒋常文,陈森洲等.BDNF对星形胶质细胞的促增殖效应及其ERK信号转导机制[J].中国老年学杂志,2007,7(27):1358-1360.
46.Song HJ,Stevens CF,Gage FH.Neural stem cells from adult hippocampus develop essential properties of functional CNS neurons[J].Nat Neurosci,2002,5(5):438-445.
47.York RD,Yao H,Dillon T,et al.Rapl mediates sustained MAP kinase activation induced by nerve growth factor[J].Nature,1998,392(6676):622-626.
48.Zhifeng Xiao,Yaxian Kong,Shufa Yang,et al.Upregulation of Flk-1 by bFGF via the ERK pathway is essential for VEGF-mediated promotion of neural stem cell proliferation [J]. Cell Research, 2007,17(1): 73-79.
49. Miguel A. Lopez-Toledano, Carolina Redondo, Maria V.T, et al. Paino and Eulalia Bazan.Tyrosine Hydroxylase Induction by Basic Fibroblast Growth Factor and Cyclic AMP Analogs in Striatal Neural Stem Cells: Role of ERK1/ERK2 Mitogen- activated Protein Kinase and Protein Kinase C [J].Histochemistry & Cytochemistry, 2004, 52(9): 1177-1189.
2.裴丽君,李竹.还原叶酸载体基因(RFCI)与神经管和颅面畸形病因学关系的研究进展[J].遗传,2004,26(2):239-243.
3.Craciunescu CN,Brown EC,Mar MH,et al.Folic Acid Deficiency During Late Gestation Decreases Progenitor Cell Proliferation and Increases Apoptosis in Fetal Mouse Brain[J].Nutr,2004,134(1):162-166.
4.王树华,单培彦.叶酸、Vitamin B12与血管性痴呆[J].卒中与神经疾病.2005,12(5):311-313.
5.Oishi K,Kamakura S,Isazawa Y,et al.Notch promotes survival of neural precursor cells via mechanisms distinct from those regulating neurogenesis[J].Development biology,2004,276(1):172-184.
6.Jin Y,Yan EZ,Fan Y,et al.Sodium ferulate prevents amyloid-beta-induccd neurotoxicity through suppression of p38 MAPK and upregulation of ERK-1/2and Akt/proteinkinase B in rat hippocampus[J].Acta Pharmacologica Sinica,2005,26(8):943-951.
7.Galabova-Kovacs G,Matzen D,Piazzolla D,et al.Essential role of B-Raf in ERK activation during extraembryonic development[J].PNAS,2006,103(5):1325-1330.
8.Reynolds BA,Weiss S.Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system[J].Science,1992,225(5052):1707-1710.
9.Eriksson PS,Perfilieva E,Bjork-Eriksson T,et al.Neurogenesis in the adult human hippocampus[J].Nat Med,1998,4(11):1207-1313.
10.Thomson JA,Itskovitz-Eldor J,Shapiro SS,et al.Embryonic stem cell lines derived from human blastocysts[J].Science,1998,282(5391):1145-1147.
11.Toda H,Takahashi J,Iwakami N,et al.Grafting neural stem cells improved the impaired spatial recognition in ischemic rats[J].Neurosci Lett,2001,316:9-12.
12.陈丽.神经干细胞在神经内科疾病中的应用[J].中国城乡企业卫生,2004,6.28.
13. Grandbarbe L, Bouissac J, Rand M, et al. Delta-Notch signaling controls the generation of neurons/glia from neural stem cells in a stepwise process[J]. Development, 2003,130 (7):1391-1402.
14. Lutolf S, Radtke F, Aguet M, et al. Notchl is required for neuronal and glial differentiation in the cerebellum[J]. Development, 2002,129(2):373-385.
15. Tanigaki K, Nogaki F, Takahashi J, et al. Notchl and Notch3 instructively restrict bFGF-responsive multipotent neural progenitor cells to an astroglial fate [J]. Neuron, 2001, 29(1):45-55.
16. Daniel Rueda, Beatriz Navarro, Alberto Martinez-Serrano, et al. The Endocannabinoid Anandamide Inhibits Neuronal Progenitor Cell Differentiation through Attenuation of the Rap1/B-RaffERK Pathway [J]. Biol Chem, 2002, 277(48):46645-466650.
17. Uyttendaele H, Marazzi G, Wu G, et al. Notch4/int-3, a mammary proto-oncogene, is an endothelial cell-specific mammalian Notch gene[J]. Development, 1996,122(7):2251-2259.
18. Artavanis-Tsakonas S, Rand MD, Lake RJ. Notch signaling: cell fate control and signal integration in development[J]. Science, 1999, 284(5415):770-776 .
19. Iso T, Kedes L and Hamamori Y. HES and HERP families: multiple effectors of the Notch signaling pathway [J]. Cell Physiol, 2003,194(3):237-255.
20. Gaiano N, Fishell G The role of Notch in promoting glial and neural stem cell fates [J]. Annu Rev Neurosci, 2002, 25:471-490.
21. Gaiano N, Nye JS, Fishell G. Radial glial identity is promoted by Notchl signaling in the murine forebrain [J]. Neuron, 2000, 26(2):395-404.
22. Nam Y, Weng AP, Aster JC, et al. Structural requirements for assembly of the CSL, intracelular Notchl.Mastermind like transcriptional activation complex [J]. Biol Chem, 2003, 278(23):21232-21239.
23. Chambers CB, Peng Y, Nguyen H, et al. Spatiotemporal selectivity of response to Notchl signals in mammalian forebrain precursors [J]. Development, 2001, 128(5):689-702.
24. Ohtsuka T. Ishibashi M. Gradwohl G. et al. Hesl and Hes5 as notch effectors in mammalian neuronal differentiation[J]. EMBO J. 1999, 18:2196-2207.
25. Appel B, Eisen JS. Regulation of neuronal specification in the zebrafish spinal cord by Delta function[J]. Development, 1998.125:371-380.
26. Berezovska O, McLe an P, Knowles R, et al. Notchl inhibits neurite outgrowth in postmitotic primary neurons[J]. Neuroscience, 1999, 93:433-439.
27. Hitoshi S, Alexson T, Tropepe V. Notch pathway molecules are essential for the maintenance, but not the generation of mammalian neural stem cells[J]. Genes Dev, 2002, 16:846-858.
28. Toshiyuki Ohtsuka, Mkoto Ishibashi, Gerald Gradwohl et al. Hesl and Hes5 as Notch effectors in mammalian neuronal differentiation[J]. EMBO, 1999, 18(8):2196-2207.
29. Stephen T. Crews. Control of cell lineage-specific development and transcription by bHLH-PAS proteins[J]. Cold Spring Harbor Laboratory Press, 2008, 4:607-620.
30. Toshiyuki Ohtsuka, Masami Sakamoto, Francois Guillemot, et al. Roles of the Basic Helix-Loop-Helix Genes Hesl and Hes5 in Expansion of Neural Stem Cells of the Developing Brain[J]. Biological Chemistry, 2001,276(32):30467— 30474.
31. Farah MH, Olson JM, Sucic HB, et al. Generation of neutons by transient expression of neural bHLH proteins in mammaliancels[J]. Development, 2000, 127: 693-702.
32. Sun Y, NadM-Vicens M, Misono S, et al. Neurogenin promotes neurogenesis and inhibits glial differentiation by independent mechanisms[J]. Cell, 2001, 104: 365-376.
33. Nieto M, Schuurmans C, Britz O, et al. Neural bHLH genes control the neuronal versus glial fate decision in cortical progenitors [J]. Neuron, 2001, 29: 401-413.
34. Bylund M, Andersson E, Novitch BG, et al. Vertebrate neurogenesis is counteracted by Soxl-3 activity[J]. Nat Neurosci, 2003, 6:1162-1168.
35. Llado J, Haenggeli C, Maragakis NJ, et al. Neural stem cells protect against glutamate-induced excitotoxicity and promote survival of injured motor neurons through the secretion of neurotrophic factors[J]. Mol Cell Neurosci, 2004, 27(3): 322-331.
36.朱春梅,王天有.营养性巨幼红细胞性贫血的防治[J].中国实用乡村医生杂志,2004,11(4):9-10.
37.Tamura T,Picciano MF.Folate and human reproduction[J].Am J Clin Nutr,2006,83(5):993-1016.
38.Snowdon DA,Greiner LH,Mortimer JA,et al.Brain infarction and the clinical expression of Alzheimer disease.The Nun Study[J].JAMA,1997,277(10):813-817.
39.Zoccolella S,Lamberti P,Iliceto G,et al.Plasma homocysteine levels in L-dopa-treated Parkinson's disease patients with cognitive dysfunctions[J].Clin Chem Lab Med,2005,43(10):1107-1110.
40.Ravaglia G,Forti P,Maioli F,et al.Homocysteine and folate as risk factors for dementia and Alzheimer disease[J].Am J Clin Nutr,2005,82(3):636-643.
41.Mattson MP and Haberman F.Folate and Homocysteine Metabolism:Therapeutic Targets in Cardiovascular and Neurodegenerative Disorders[J].Curr Medl Chem,2003,10(19):1923-1929.
42.黄国伟,刘欢.叶酸、维生素B_6、B_(12)对脑缺血大鼠学习记忆的影响[J].中华预防医学杂志,2007,41(3):212-214.
43.Bottiglieri T,Laundy M,Crellin R,et al.Homocysteine,folate,methylation and monomunine metabolism in depression[J].J Neurol Neurosurg Psychiatry,2000,69:228-232.
44.Alpert JE,Mischoulon D,Nierenherg AA,et al.Nutrition and depression:focus on folate[J].Nutrition,2000,16:544-546.
45.Fava M,BoresJS,Alpert JE,et al.Folate,vitamin B_(12)and homocysteine in major depressive disorder[J].Am J Psychiatry,1997,154:426-428.
46.Kruman Ⅱ,Mouton PR,Emokpae R Jr,et al.Folate deficiency inhibits proliferation of adult hippocampal progenitors[J].Neuroreport,2005,16(10):1055-1059.
47.Shea TB,Rogers E.Folate quenches oxidative damage in brains of apolipoprotein E-deficient mice:augmentation by vitamin E[J].Brain Res Mol Brain Res,2002,108(1-2):1-6.
48.Ho PI,Collins SC,Dhitavat S,et al.Homocysteine potentiates beta-amyloid neurotoxicity,role of oxidative stress[J].Neurochem,2001,78(2):249-253.
49.Kruman Ⅱ,Kumaravel TS,Lohani A,et al.Folic acid deficiency and homocysteine impair DNA repair in hippocampal neurons and sensitize them to amyloid toxicity in experimental models of Alzheimer's disease[J].Neurosci,2002,22(5):1752-1762.
50.Keating E,Lemos C,Azevedo I,et al.Comparison of folic acid uptake characteristics by human placental choriocarcinoma cells at acidic and physiological Ph[J].Can J Physiol Pharmacol,2006,84(2):247-55.
51.张琦,郑志竑.Notch信号通路与神经干细胞的增殖分化[J].国外医学遗传学分册,2004,27(2):68-71.
52.Wada T,Penninger J M.Mitogen-activated protein kinases in apoptosis regulation[J].Oncogene,2004,23(16):2838-2849.
53.Seger R,Krebs E G.The MAPK signaling cascade[J].FASEB J,1995,9(9):726-735.
54.Ping Hu,Zhang Han,Anthony D.et al.Critical Role of Endogenous Akt/IAPs and MEK1/ERK Pathways in Counteracting Endoplasmic Reticulum Stress-induced Cell Death[J].Biological chemistry,2004,279(47):49420-49429.
55.Campos LS,Leone DP,Relvas JB,et al.Betal integrins activate a MAPK signalling pathway in neural stem cells that contributes to their maintenance[J].Development,2004,131(14):3433-3444.
56.Eunhae Kim,Amy L.Clark,et al.Belcheva.μ-and κ-Opioids Induce the Differentiation of Embryonic Stem Cells to Neural Progenitors[J].Biological Chemistry,2006,281(44):33749-33760.
57.端木德强,李中海,王敬泽.MAPK信号转导通路研究进展[J].细胞生物学杂志,2002,24(3):151-155.
58.姜勇,龚小卫.MAPK信号转导通路对炎症反应的调控[J].生理学报,2002,52(4):267-271.
59.牟金叶,陈晓光.促分裂原激活的蛋白激酶(MAPK)信号转导通路的研究进展[J].生命科学,2002,14(4):208-21.
60.张璐,姜勇,张琳.丝裂原活化蛋白激酶信号转导通路研究进展[J].国外医 学(生理、病理科学与临床分册),1999,19(2):84-87.
61.Adachi T,Kar S,Wang M,et al.Transient and sustained ERK phosphorylation and nuclear translocation in growth control[J].J Cell Physiol,2002,192(2):151-159.
62.孙小妹,毛萌,周晖等.脑源性神经营养因子对缺氧胚脑皮质神经元发挥保护作用的细胞内信号传导机制[J].华西医学,2006,21(3):454-457.
63.王彪,邢伟,许金华等.铝对大鼠海马ERK蛋白及mRNA表达影响[J].中国公共卫生,2006,22(12):1464-1466.
64.Vartiainen N,Goldsteins G,Keksa-Goldsteine V,et al.Aspirin Inhibits p44/42Mitogen-Activated Protein Kinase and Is Protective Against Hypoxia/Reoxygenation Neuronal Damage[J].Stroke,2003,34(3):752-757.
65.Madan P,Calder MD,Watson AJ.Mitogen-activated protein kinase(MAPK)blockade of bovine preimplantation embryogenesis requires inhibition of both p38 and extracellular signal-regulated kinase(ERK)pathways[J].Reproduction,2005,130(1):41-51.
66.Song HJ,Stevens CF,Gage FH.Neural stem cells from adult hippocampus develop essential properties of functional CNS neurons[J].Nat Neurosci,2002,5(5):438-445.
67.York RD,Yao H,Dillon T,et al.Rapl mediates sustained MAP kinase activation induced by nerve growth factor[J].Nature,1998,392(6676):622-626.
68.Zhifeng Xiao,Yaxian Kong,Shufa Yang,et al.Upregulation of Flk-1 by bFGF via the ERK pathway is essential for VEGF-mediated promotion of neural stem cell proliferation[J].Cell Research,2007,17(1):73-79.
69.Miguel A.Lopez-Toledano,Carolina Redondo,et al.Tyrosine Hydroxylase Induction by Basic Fibroblast Growth Factor and Cyclic AMP Analogs in Striatal Neural Stem Cells:Role of ERK1/ERK2 Mitogen-activated Protein Kinase and Protein Kinase C[J].Histochemistry & Cytochemistry,2004,52(9):1177-1189.
70.Borsello T,Clarke PG,Hirt L,et al.A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia[J].Nat Med,2003,9(9):1180-1186.
71.Ferrer I,Friguls B,Dalfo E,et al.Early modifications in the expression of mitogen-activated protein kinase(MAPK/ERK), stress-activated kinases SAPK/JNK and p38, and their phosphorylated substrates following focal cerebral ischemia[J]. Acta Neuropathol(Bed), 2003,105(5):425-437.
72. HuB, LiuC, Bramlett H, et al. Changes in trkB-ERK1/2-Creb/Elk-l pathways in hippocampal mossy fiber organization after traumatic brain injury[J]. J Cereb Blood Flow Metab, 2004,24(8):934-943.
73. Swatton JE, Sellers LA, Faun RL, 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.
74. Raghupathi R, Muir J K, Fulp C T, et al. Acute activation of mitogen-activated protein kinases following traumatic brain injury in the rat: implications for posttraumatic cell death[J]. Exp Neurol, 2003,183(2):438-448.
75. Raghupathi R, Graham DI, Mclntosh TK. Apoptosis after traumatic brain injury[J]. J Neurotrauma, 2000,17(10):927-938.
76. Marie-Therese Stockhausen, Jonas Sjolund and Hakan Axelson. Regulation of the Notch target gene Hes-1 by TGFa induced Ras/MAPK signaling in human neuroblastoma cells [J]. Experimental Cell Research, 2005,310 (1): 218-228.
1. Artavanis-Tsakonas S,Rand MD, Lake RJ. Notch signaling: cell fate control and signal integration in development[J]. Science, 1999,284:770-776.
2. Allman D, Aster JC, Pear W S. Notch signaling in hematopoiesis and early lymphocyte development[J]. Immunol Rev, 2002,187:75-86.
3. Arias A, Zecchini V, Brennan K. CSL-independent Notch signalling:a checkpoint in cell fate decisions during development[J]? Curr Opin Genet Dev. 2002,12:524-533.
4. Kojika S, Grifin JD. Notch receptors and hematopoiesis[J]. Exp Hematol, 2001, 29:1041-1052.
5. Lai EC. Notch cleavage:Nieastrin helps Presenilin make the final cut[J]. Curr Biol. 2002,12: R200-R202.
6. Milner LA, Bigas A. Notch as a mediator of cell fate determination in hema-Topoiesis: evidence and speculation[J]. Blood, 1999, 93: 2431-2448.
7. Bray S, Furriols M. Notch pathway: making sense of suppressor of hairless[J]. Curt Biol, 2001, 11 :R217-R221.
8. Okajima T, Irvine KD. Regulation of notch signaling by olinked fucose[J], Cell. 2002,111:893-904.
9. Shi S, Stanley P. Protein O-fueosyhransferasel is an essential component of Notch signaling pathways[J]. Proc Natl Acad Sci USA. 2003, 100: 5234-5239.
10. Lin SE, Oyama T, Nagase T, et al. Identification of new human mastermind proteins defines a family that consists of positive regulators for Notch signaling[J]. J Biol Chem, 2002,277: 50612-50620.
11. Jafar-Nejad H, Norga K, Bellen H. Numb:"Adapting" Notch for Endocytosis[J]. Dev Cell. 2002,3:155-156.
12. Yamamoto N, Yamamoto S, Inagaki F, et al .Role of Deltex-1 as a transcriptional regu lator downstream of the Notch receptor[J]. J Biol Chem, 2001, 276:45031-45040.
13. Matsuno K, Diederich RJ, Go MJ, et al. Dehex acts as a positive regulator of Notch signaling through interactions with the Notch ankyrin repeats[J]. Development,1995,121:2633-2644.
14.Matsuno K,Ito M,Hori K,et al.Involvement of a praline-rich motif and RING-H2 finger of Dehex in the regulation of Notch signaling[J].Development,2002,129:1049-1059.
15.Sestan N,Artavanis-Tsakonas S,Rakic P.Contact-de-pendent inhibition of cortical neurite growth mediated by Notch signaling[J].Science,1999,286:741-746.
16.Izon DJ,Aster JC,He Y,et al.Dehexl redirects lymphoid progenitors to the B cell lineage by antagonizing Notchl[J].Immunity,2002,16:231-243.
17.Ravaglia G,Forti P,Maioli F,Martelli M,Servadei L,Brunetti N,Porcellini E,Licastro F.Homocysteine and folate as risk factors for dementia and Alzheimer disease[J].Am J Clin Nutr,2005,82(3):636-643.
18.Mattson MP and Haberman F.Folate and Homocysteine Metabolism:Therapeutic Targets in Cardiovascular and Neurodegenerative Disorders[J].Curr Medl Chem,2003,10(19):1923-1929.
19.黄国伟,刘欢.叶酸、维生素B_6、B_(12)对脑缺血大鼠学习记忆的影响[J].中华预防医学杂志,2007,41(3):212-214.
20.Hitoshi S,Alexson T,Tropepe V,Donoviel D,Elia AJ,Nye JS,Conlon RA,Mak TW,Bernstein A,van der Kooy D.Neural stem cell lineages are regionally specified,but not committed,within distinct compartments of the developing brain[J].Development,2002,129(1):233-244.
21.王海燕,徐如祥,姜晓丹.Notch信号转导通路对神经干细胞的调控[J].生命的化学,2003,23(3):167-169.
22.Tsai RY,McKay RD.Cell contact regulates fate choice by cortical stem cells[J].J Neurosci,2000,20(10):3725-3735.
23.Alexson TO,Hitoshi S,Coles BL,et al.Notch signaling is required to maintain all neural stem cell populations irrespective of spatial or temporal niche[J].Dev Neurosci,2006,28(1-2):34-48.
24.张琦,郑志竑.Notch信号通路与神经干细胞的增殖分化[J].国外医学遗传学分册,2004,27(2):68-71.
25.刘素芳.Notch信号与神经干细胞分化[J].河南医学研究,2004,13(4),366-368.
26. Johnson JE, Biren SJ, Anderson DJ. Two rat homologues of drosophila achaete-scute specifically expressed in neuronal precursors[J]. Nature, 1990, 346(6287):858-861.
27. Douziech N, Calvo E, Laine J, et al. Activation of MAP kinases in growth[J]. Cell Signal, 1999,11:591-602.
28. Rampoldi, L, Zimbello R, Bortoluzzi S, et al. Chromosomal localization of four MAPK signaling cascade genes: MEK1, MEK3, MEK4 and MEKK5[J]. 1997, Cytogenetics and Cell Genetics[J], 78:301-303.
29. VitNe, G. Fagligt program - parallelle sesjoner[J].1999, Journal of Applied Microbiology, 87:288-293.
30. Canagarajah, B. J, et al. Activation mechanism of the MAP kinase ERK2 by dual phosphorylation[J]. 1997, Cell, 90:859-869.
31. Andrei, V. Khokhlatchev, Bertram Canagarajah, Julie Wilsbacher, et al. Activation Mechanism of the MAP Kinase ERK2 by Dual Phosphorylation[J]. 1998, Cell, 93:605-615.
32. Maria Deak, Andrew D, Clifton, John M, Lucocq et al. Mitogen- and stress-activated protein kinase-1 (MSK1) is directly activated by MAPK and SAPK2/p38, and may mediate activation of CREB[J]. 1998, EfBO, 17(15):4426-4441.
33. Reiko Sugiura, Takashi Toda, Susheela Dhut, et al. The MAPK kinase Pekl acts as a phosphorylation-dependent molecular switch[J]. 1999, Letters to nature, 399:479- 482.
34. SHUICHAN Xu, David Robbins, Jeff Frost, et al. MEKK1 phosphorylates MEK1 and MEK2 but does not cause activation of mitogen-activated protein kinase[J]. 1995, PNAS, 92: 6808-6812.
35. Borsello T, Clarke PG, Hirt L, et al. A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia[J]. Nat Med. 2003, 9(9):1180-1186.
36. Ferrer I, Friguls B, Dalfo E, et al. Early modifications in the expression of mitogen-activated protein kinase(MAPK/ERK), stress-activated kinases SAPK/JNK and p38, and their phosphorylated substrates following focal cerebral ischemia[J].Acta Neuropathol(Bed),2003,105(5):425-437.
37.HuB,LiuC,Bramlett H,et al.Changes in trkB-ERK1/2-cREB/Elk-1 pathways in hippocampal mossy fiber organization after traumatic brain injury[J].J Cereb Blood Flow Metab,2004,24(8):934-943.
38.Swatton JE,Sellers LA,Faun RL,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.
39.Raghupathi R,Muir J K,Fulp CT,et al.Acute activation of mitogen-activated protein kinases following traumatic brain injury in the rat:implications for posttraumatic cell death[J].Exp Neurol,2003,183(2):438-448.
40.Raghupathi R,Graham DI,Mclntosh TK.Apoptosis after traumatic brain injury[J].J Neurotrauma,2000,17(10):927-938.
41.孙小妹,毛萌,周晖等.脑源性神经营养因子对缺氧胚脑皮质神经元发挥保护作用的细胞内信号传导机制[J].华西医学,2006,21(3):454-457.
42.顾平,王彦永,马芹颖等.ERK1/2信号转导通路对骨髓基质细胞条件培养液诱导中脑神经干细胞分化作用的影响[J].2007,28(6):589-592.
43.Almeida RD,Manadas BJ,Melo CV,et al.Neuroprotection by BDNF against glutamate-induced apoptotic cell death is mediated by ERK and PI3-kinase pathways[J].Cell Death Difer,2005,12(10):1329-43.
44.Sakamoto T,Kawasoe Y,Shen JS,et al.Adenoviral gene transfer of GDNF,BDNF an d TGF beta 2,but not CNTF,cardiotrophin-1 or IGF1,protects injured adult motoneurons after facial nerve avulsion[,J].J Neuroeci Res,2003,72(1):54-64.
45.蒋常文,陈森洲等.BDNF对星形胶质细胞的促增殖效应及其ERK信号转导机制[J].中国老年学杂志,2007,7(27):1358-1360.
46.Song HJ,Stevens CF,Gage FH.Neural stem cells from adult hippocampus develop essential properties of functional CNS neurons[J].Nat Neurosci,2002,5(5):438-445.
47.York RD,Yao H,Dillon T,et al.Rapl mediates sustained MAP kinase activation induced by nerve growth factor[J].Nature,1998,392(6676):622-626.
48.Zhifeng Xiao,Yaxian Kong,Shufa Yang,et al.Upregulation of Flk-1 by bFGF via the ERK pathway is essential for VEGF-mediated promotion of neural stem cell proliferation [J]. Cell Research, 2007,17(1): 73-79.
49. Miguel A. Lopez-Toledano, Carolina Redondo, Maria V.T, et al. Paino and Eulalia Bazan.Tyrosine Hydroxylase Induction by Basic Fibroblast Growth Factor and Cyclic AMP Analogs in Striatal Neural Stem Cells: Role of ERK1/ERK2 Mitogen- activated Protein Kinase and Protein Kinase C [J].Histochemistry & Cytochemistry, 2004, 52(9): 1177-1189.