摘要
第一部分PGG对抗MPP+孵育所致PC12细胞的氧化应激反应
摘要目的：研究1,2,3,4,6-五-氧位-没食子酰基-β-右旋葡萄糖(1,2,3,4, 6-penta-O-galloy-β-D-glucose, PGG)诱导PC12细胞中血红素加氧酶(heme oxygenase,HO-1)的产生及HO-1对抗MPP+孵育PC12所产生的氧化应激反应。方法：用不同浓度的(0、5、20、50、80、100μM) PGG处理经过神经营养因子(nerve growth factor—NGF)孵育一周的PC12细胞10小时,以及用30μM的PGG处理经过NGF孵育过一周的PC12不同时间。观察HO-1mRNA变化时所用的时间点为0、3、6、12、18、24h,观察HO-1蛋白变化时所用的时间点为-0、5、10、15、20、30h；通过免疫印迹法(Western blotting)、RT-PCR、活性氧试剂盒检测以及MTT方法观察PC12细胞中HO-1 mRNA和蛋白的合成与PGG作用浓度与作用时间的关系,PC12细胞中HO-1活力与PGG作用浓度与作用时间的关系,以及PGG对抗由于孵育MPP+所致的对PC12细胞产生的氧化应激反应。结果：PGG能促进PC12细胞中HO-1mRNA和蛋白的合成,HO-1mRNA和蛋白的合成呈浓度和时间依赖性,在PGG作用10h下,20gM的PGG便能增加HO-1mRNA和蛋白的合成,50μMPGG作用下HO-1mRNA和蛋白的合成呈现高峰,之后HO-1mRNA和蛋白合成量下降,但仍高于20μM PGG作用时HO-1mRNA和蛋白的合成量；在30μMPGG作用下,PC12细胞中HO-1mRNA和蛋白的合成均呈时间依赖性,HO-1mRNA在6h时合成增多,12h达高峰,HO-1蛋白在10h时合成增多,15时达高峰,高峰后的时间点HO-1mRNA合成量仍大于6h点时HO-1mRNA合成量,蛋白的合成量仍大于10h点时的HO-1的蛋白量。HO-1的活性变化也呈现浓度与时间依赖性,其趋势与HO-1蛋白表达的浓度与时间依赖性一致。定量与定性上均观察到PGG抑制活性氧簇(reactive oxygen species, ROS)的产生。结论：PGG促进PC12细胞中HO-1mRNA和蛋白的合成,在PGG的作用下,PC12细胞中HO-1mRNA和蛋白的合成呈现浓度和时间依赖性,HO-1的活力变化也呈现浓度和时间依赖性。PGG可通过诱导HO-1的产生对抗MPP+对PC12产生的氧化应激反应。
第二部分PGG对ERKl/2信号通路诱导HO-1的影响
目的：本部分研究细胞外信号调节酶1/2(extracellular singnal-regulated kinase, ERK1/2)信号通路在1,2,3,4,6-五-氧位-没食子酰基(1,2,3,4,6-penta-O-galloy-β-D-glucose,PGG)诱导PC12细胞中HO-1产生中的作用。方法：将细胞分成不同的组别,用PGG、MPP+,PD98059以及ZnPP处理PC12细胞,通过Western blotting和MTT方法,观察药物处理前后PC12细胞中ERK1/2磷酸化程度,Nrf2核转位、HO-1的表达以及细胞活力的变化。结果：PGG可促进PC12细胞中ERK1/2磷酸化增高,增高的ERK1/2磷酸化可促进PC12细胞细胞浆中的Nrf2向细胞核转位,从而促进了HO-1的合成增多,达到保护MPP+所损害的PC12细胞的作用；PD98059可通过抑制ERK1/2磷酸化而部分的抑制PC12细胞的中HO-1的表达,从而部分抑制了PGG对MPP+所损伤的PC12的保护作用。结论：ERK1/2信号通路在PGG保护MPP+损害PC12细胞中发挥重要的作用。
第三部分PGG对PI3K/AKT信号通路诱导HO-1的影响
目的：本部分研究磷脂酰肌醇-3激酶(P13K)／蛋白激酶AKT(phosphatidylinositol-3 kinase/protein AKT, PI3K/AKT)信号通路在PGG促进HO-1表达中的作用。方法：将细胞分成不同的组别,用PGG、MPP+, LY294002以及ZnPP处理PC12细胞,通过Western blotting和MTT方法,观察PC12细胞中AKT磷酸化程度、Nrf2核转位程度、HO-1的表达以及细胞活力的变化。结果：PGG可促进PC12细胞中AKT磷酸化增高,增高的AKT磷酸化可促进PC12细胞胞浆中的Nrf2向细胞核转位,从而促进了HO-1的合成增多,达到保护MPP+所损害的PC12细胞的作用；LY294002可通过抑制AKT磷酸化而部分的抑制PC12细胞的中HO-1的表达,从而部分抑制了PGG对MPP+所损伤的PC12的保护作用。并且LY294002与PD98059联合作用能完全阻断Nrf2的核转位与HO-1的表达,但无法完全消除PGG对与MPP+孵育PC12细胞的保护作用。结论：PI3K/AKT信号通路在PGG保护MPP+损害PC12细胞中发挥重要的作用。
Part I PGG ameliorates oxidative stress.induced byMPP+in PC 12 cells
Object:To investigate whether PGG could induce the expression of HO-1 and Whether HO-1 could ameliorate oxidative stress induced by MPP+in PC 12 cells Methods:we treated the PC 12 cells,which were incubated by NGF one week before drug treatment, for 10h with various concentrations of PGG and treated the PC 12 cells, which were incubated by NGF one week before drug treatment,for various time points with 30μM PGG;we used these various time points-0、3、6、12、18、24h when we studied the production of HO-1 mRNA, and we used these various time points-0、5、10、15、20、30h when we studied the expression of HO-1 protein.Then we examined the relationship between various concentrations and various time points of PGG and the production of HO-1 mRNA and protein by Western blotting, RT-PCR, DCFH-DA and MTT assay. the relationship between activity of HO-1 and various concentrations and various time points of PGG; and whether PGG could ameliorate oxidative stress.induced by MPP+in PC12 cells.Results:The production of HO-1 mRNA and protein is in dose-dependent concentration-dependent manners. HO-1 mRNA levels showed an initial rise at 20μM,a peak induction at 50μM,and a decrease by 80μM. Induction of the HO-1 mRNA in PC12 cells is a time-dependent manner, HO-1 mRNA levels showed an initial rise at 6h,a peak induction at 12h,followed by a decrease at 18h. Exposure of PC 12 cells to various concentrations of PGG for 10h resulted in a concentration-dependent increase in HO-1 protein expression,and the increase trend is similar to production of HO-1 mRNA.And the trend of HO-1 protein levels showed an initial rise atlOh,a peak induction at 15h,followed by a decrease at 20h. HO-1 activity was also increased in dose-dependent and time-dependent manners which is similar to the dose-dependent and time-dependent manners of HO-1 protein expression. And PGG could ameliorate induced byMPP+in PC 12 cells.
Part II Effect of PGG on the expression of HO-1 via ERK1/2 signaling pathway
Object:Here, we study the role of ERK1/2 signaling pathway in induction of HO-1 by PGG in the PC 12 cells.
. Methods:The PC12 cells were divided into different groups,and then were incubated by PGG, PD98059 and ZnPP according to different groups. And then Western blotting and MTT assay were used to investigate the difference of p-ERK1/2,Nrf2、HO-1 expression,and activity of the PC 12 cells after the treatment with different drugs.Results: PGG could promote the phosphorylation of ERK1/2 which promoted Nrf2 nuclear translocation which promoted the expression of HO-1 and thus protected PC 12 from damage by MPP+.But PD98059 produced opposite role in the process that PGG protected PC 12 cells-PD98059 suppressed the protection of PC 12 cells by PGG partly.
Conclusion:ERK1/2 signaling pathway played a role in the protection of PC12 cells by PGG.
PartⅢEffect of PGG on the expression of HO-1 via PI3K/AKT signaling pathway
Object:Here, we study the role of PI3K/AKT signaling pathway in induction of HO-1 by PGG in the PC 12 cells.
Methods:The PC12 cells were divided into different groups,and then were incubated by PGG, PD98059 and ZnPP according to different groups. And then Western blotting and MTT assay were used to investigate the difference of p-AKt,Nrf2、HO-1 xpression,and activity of the PC 12 cells after the treatment with different drugs.
Results:PGG could promote the phosphorylation of AKt which promoted Nrf2 nuclear translocation which promoted the expression of HO-1 and thus protected PC 12 from damage by MPP+。But LY294002 produced opposite role in the process that PGG protected PC 12 cells-LY294002 suppressed the protection of PC 12 cells by PGG partly.LY294002 and PD98059 together could blocked Nrf2 nuclear translocation and expression of HO-1 completely, but could not abolish completely the protection of PC 12 cells by PGG.
Conclusion:PI3K/AKT signaling pathway played a role in the protection of PC 12 cells by PGG.
引文
[1]Katori M, Busuttil RW, Kw JW. Heme oxygenase-1 system in organ transplantation. Transplantation 2002; 74:905-912.
[2]Fu WP, Zhao ZH, Fang LZ, Sun C, Liu L, Zhang JQ, et al. Heme oxygenase-1 polymorphism associated with severity of chronic obstructive pulmonary disease. Chin Med J 2007; 120:12-16.
[3]Li P, Elrayess MA, Gomma AH, Palmen J, Hawe E, Fox KM, et al. The microsatellite polymorphism of heme oxygenase-lis associated with baseline plasma IL-6 level but not with restenosis after coronary in-stenting. Chin Med J 2005;118:
[4]Zhang J,Li L, Kim SH, et al.Anti-cancer, anti-diabetic and other pharmacologic and biological activities of penta-galloyl-glucose.Pharm Res 2009,26(9):2033-80.
[5]Byung-Min Choia, Hyung-Jin,et al.1,2,3,4,6-Penta-O-galloyl-beta-d-glucose protects rat neuronal cells(Neuro 2A) from hydrogen peroxide-mediated cell death via the induction of heme oxygenase-1.Neuroscience Letters 2002.328(185-189)
[6]Tenhunen, R, Marver,H.S. and Schmid,R., The enzymatic catabolism of hemoglobin stimulation of microsomal heme oxygenase by hemin, J. Lab. Clin. Med,75(1970) 410-421.
[7]Shoskes DA, Halloran PF. Delayed graft function in renal transplantation:etiology, management and long-term significance. J Urol 1996; 155:1831-1840.
[8]Katori M, Busuttil RW, Kw JW. Heme oxygenase-1 system in organ transplantation. Transplantation 2002; 74:905-912.
[9]Fu WP, Zhao ZH, Fang LZ, Sun C, Liu L, Zhang JQ, et al. Heme oxygenase-1 polymorphism associated with severity of chronic obstructive pulmonary disease. Chin Med J 2007; 120:12-16.
[10]Li P, Elrayess MA, Gomma AH, Palmen J, Hawe E, Fox KM, et al. The microsatellite polymorphism of heme oxygenase-lis associated with baseline plasma IL-6 level but not with restenosis after coronary in-stenting. Chin Med J 2005; 118:1525-1532
[11]Neto JS, Nakao A, Kimizuka K, Romanosky AJ, Stolz DB, Uchiyama T, et al. Protection of transplant-induced renal ischaemic reperfusion injury with carbon monoxide. Am J Physiol Renal Physiol 2004; 287:979-989.)
[12]Nishizawa M, Yamagishi T, Nonaka G, Nishioka I, Bando H. Novel Hydrolyzable Tannins from Nuphar-Japonicum Dc. Chemical & Pharmaceutical Bulletin 1982;30(3):1094-7.)。
[13]Oh GS, Pae HO, Oh H, et al. In vitro anti-proliferative effect of 1,2,3,4,6-penta-O-galloyl-beta-D-glucose on human hepatocellular carcinoma cell line, SK-HEP-1 cells. Cancer Lett 2001;174(1):17-24. [PubMed:11675148]
[14]Huh JE, Lee EO, Kim MS, et al. Penta-O-galloyl-beta-D-glucose suppresses tumor growth via inhibition of angiogenesis and stimulation of apoptosis:roles of cyclooxygenase-2 and mitogen-activated protein kinase pathways. Carcinogenesis 2005;26(8):1436-45. [PubMed:15845650]
[15]Ren YM, Chen XZ. Distribution, Bioactivities and Therapeutical Potentials of Pentagalloylglucopyranose. Current Bioactive Compounds 2007;3:81-9.)
[16]Hu H, Lee HJ, Jiang C, et al. Penta-1,2,3,4,6-O-galloyl-beta-D-glucose induces p53 and inhibits STAT3 in prostate cancer cells in vitro and suppresses prostate xenograft tumor growth in vivo. Mol Cancer Ther 2008;7(9):2681-91. [PubMed:18790750]
[17]Nakagawa H, Hasumi K, Woo JT, Nagai K, Wachi M. Generation of hydrogen peroxide primarily contributes to the induction of Fe(Ⅱ)-dependent apoptosis in Jurkat cells by (-)-epigallocatechin gallate. Carcinogenesis 2004;25(9):1567-74. [PubMed: 15090467]
[18]Lee KW, Hur HJ, Lee HJ, Lee CY. Antiproliferative effects of dietary phenolic substances and hydrogen peroxide. Journal of agricultural and food chemistry 2005;53(6):1990-5. [PubMed:15769125]
[19]Okubo T, Nagai F, Seto T, Satoh K, Ushiyama K, Kano I. The inhibition of phenylhydroquinone-induced oxidative DNA cleavage by constituents of Moutan Cortex and Paeoniae Radix. Biol Pharm Bull 2000;23(2):199-203. [PubMed: 10706384]
[20]Park EJ, Zhao YZ, An RB, Kim YC, Sohn DH.1,2,3,4,6-penta-O-galloyl-beta-D-glucose from Galla Rhois protects primary rat hepatocytes from necrosis and apoptosis. Planta Med 2008;74(11):1380-3. [PubMed:18622905]
[21]Dinkova-Kostova AT, Talalay P. Direct and indirect antioxidant properties of inducers of cytoprotective proteins. Mol Nutr Food Res 2008;52(Suppl1):S128-38. [PubMed:18327872]
[22]Choi BM, Kim HJ, Oh GS, et al.1,2,3,4,6-Penta-O-galloyl-beta-D-glucose protects rat neuronal cells (Neuro 2A) from hydrogen peroxide-mediated cell death via the induction of heme oxygenase-1. Neurosci Lett 2002;328(2):185-9. [PubMed: 12133584])
[1]Chiu PY, Chen N, Leong PK et al.Schisandrin B elicits a glutathione antioxidant response and protects against apoptosis via the redox-sensitive ERK/Nrf2 pathway in H9c2 cells. Mol Cell biochem 2011 April;350(1-2); 237-50.
[2]Farombi EO, Shrotriya S, Na HK, et al.Curcumin attenuates dimethylnitrosamine-induced liver injury in rats through Nrf2-mediated induction of heme oxygenase-1 Food Chem Toxicol.2008 Apr;46(4):1279-87. Epub 2007 Sep 26
[3]镇澜,刘义,吕立群,等。17p-雌二醇对子宫内膜异位症患者在位子宫内膜间质细胞ERK1/2信号转导通路活化的影响。[J]华中科技大学学报(医学版),2010,8,39(4)： 446
[4]Chan J. Y., Kwong M., Biochim. Impaired expression of glutathione synthetic enzyme genes in mice with targeted deletion of the Nrf2 basic-leucine zipper protein. Biochim Biophys Acta.2000 Dec 15; 1517(1):19-26.
[5]Ishii T., Itoh K., Takahashi S., Sato H., Yanagawa T., Katoh Y., Bannai S., Yamamoto M., Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages. J. Biol. Chem.,275,16023—16029 (2000).
[6]Shih A. Y, Li P., Murphy T. H., A small-molecule-inducible Nrf2-mediated antioxidant response provides effective prophylaxis against cerebral ischemia in vivo. J. Neurosci.,25,10321—10335 (2005).
[7]The Neuroprotection of Oxymatrine in Cerebral Ischemia/Reperfusion Is Related to Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2)-Mediated Antioxidant Response: Role of Nrf2 and Hemeoxygenase-1 Expression)
[8]Dinkova-Kostova AT, Talalay P. Direct and indirect antioxidant properties of inducers of cytoprotective proteins. Mol Nutr Food Res 2008;52(Suppl 1):S128-38. [PubMed:18327872]
[9]Choi BM, Kim HJ, Oh GS, et al.1,2,3,4,6-Penta-O-galloyl-beta-D-glucose protects rat neuronal cells (Neuro 2A) from hydrogen peroxide-mediated cell death via the induction of heme oxygenase-1. Neurosci Lett 2002;328(2):185-9. [PubMed: 12133584])
[10]Ishii T., Itoh K., Takahashi S., Sato H., Yanagawa T., Katoh Y., Bannai S., Yamamoto M., J. Biol. Chem.,275,16023—16029 (2000)
[11]Choi BM, Kim HJ, Oh GS, et al.1,2,3,4,6-Penta-O-galloyl-beta-D-glucose protects rat neuronal cells (Neuro 2A) from hydrogen peroxide-mediated cell death via the induction of heme oxygenase-1. Neurosci Lett 2002;328(2):185-9. [PubMed: 12133584])
[12]Tobiume K, Matsuzawa A, Takahashi T, Nishitoh H, Morita K, Takeda K, Minowa O, Miyazono K, Noda T, Ichijo H (2001) ASK1 is required for sustained activations of JNK/p38 MAP kinases and apoptosis. EMBO Rep 2:222-228。
[13]Chang F, Steelman LS, Shelton JG, Lee JT, Navolanic PM, Blalock WL, Franklin R, McCubrey JA (2003) Regulation of cell cycle progression and apoptosis by the Ras/Raf/MEK/ERK pathway (review). Int J Oncol 22:469-480).
[14]Yuan X, Xu C, Pan Z, Keum YS, Kim JH, Shen G, Yu S, Oo KT, Ma J, Kong AN (2006) Butylated hydroxyanisole regulates ARE-mediated gene expression via Nrf2 coupled with ERK and JNK signaling pathway in HepG2 cells. Mol Carcinog 45:841-85040.
[15]Wu CC, Hsu MC, Hsieh CW, Lin JB, Lai PH, Wung BS (2006) Upregulation of heme oxygenase-1 by epigallocatechin-3-gallate via the phosphatidylinositol 3-kinase/Akt and ERK pathways. Life Sci 78:2889-2897
[16]Na HK, Kim EH, Jung JH, Lee HH, Hyun JW, Surh YJ (2008) (-)-Epigallocatechin ERK in human mammary epithelial cells. Arch Biochem Biophys 476:171-177),
[17]Chiu PY, Chen N, Leong PK, et al 。 Schisandrin B elicits a glutathione antioxidant response and protects against apoptosis via the redox-sensitive ERK/Nrf2 pathway in H9c2 cells. Mol Cell Biochem.2011 Apr;350(1-2):237-50. Epub 2010 Dec 31
[1]镇澜,刘义,吕立群,等。17-β雌二醇对子宫内膜异位症患者在位子宫内膜间质细胞PI3K/Akt信号转导通路活化的影响。华中科技大学学报(医学版),2010,39(3)： 321
[2]Chan J. Y., Kwong M., Impaired expression of glutathione synthetic enzyme genes in mice with targeted deletion of the Nrf2 basic-leucine zipper protein. Biochim. Biophys. Acta,1517,19—26 (2000).
[3]Ishii T., Itoh K., Takahashi S., et al., Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages J. Biol. Chem.275, 16023—16029(2000).
[4]Shih A. Y., Li P., Murphy T. H., A small-molecule-inducible Nrf2-mediated antioxidant response provides effective prophylaxis against cerebral ischemia in vivo. J. Neurosci.,25,10321—10335 (2005)
[5]The Neuroprotection of Oxymatrine in Cerebral Ischemia/Reperfusion Is Related to Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2)-Mediated Antioxidant Response: Role of Nrf2 and Hemeoxygenase-1 Expression。
[6]Dinkova-Kostova AT, Talalay P. Direct and indirect antioxidant properties of inducers of cytoprotective proteins. Mol Nutr Food Res 2008;52(Suppl 1):S128-38.
[7]Choi BM, Kim HJ, Oh GS, et al.1,2,3,4,6-Penta-O-galloyl-beta-D-glucose protects rat neuronal cells (Neuro 2A) from hydrogen peroxide-mediated cell death via the induction of heme oxygenase-1. Neurosci Lett 2002;328(2):185-9.
[8]Feng J, Zhang P, Chen X, He G.PI3K and ERK/Nrf2 Pathways Are Involved in Oleanolic Acid-Induced Heme Oxygenase-1 Expression in Rat Vascular Smooth Muscle Cells.J Cell Biochem.2011 Jun; 112(6):1524-31.
[1]Levy G,Tang MX,Louis ED, et al. The association of incident dementia with mortality in PD[J]. Neurology,2002,59 (11):1708-1713.
[2]汪锡金、张煜、陈生弟,帕金森病发病机制与治疗研究十年进展[J],2010,10(1)：36-42.中国现代神经病学杂志。
[3]叶晓来、王瑛、陈生弟,帕金森病的相关因素Jou rnal of Internat ion al Neu rology and N eurosu rgery,2006,33 (5):484-487.
[4]郝斌,周晓平,帕金森病发病机制的研究,中国药物与临床[J],2004,4(7),525-527
[5]陈健,PD的病因,山东医药[J],2009,40(31),105-107。
[6]Dawson TM,Dawson VL. Molecular pathways of neurodegeneration in Parkinson's disease[J]. Science,2003,302(5646):819-822.
[7]Pankratz N, Foroud T. Genetics of Parkinson Disease [J]. NeuroRx,2004,1 (2):235-242.
[8]Giasson B I, Duda JE, Qu inn SM, et al Neuron al a lpha-synucleinopathy with severe m ovem ent d isorder in m ice expressing A53T hum an alpha-synuclein[J]Neuron,2002,34 (4:521-33.
[9]Feany MB, B ender WW.A D rosoph ila m odel of Park inson&s d isease [J] Natu re, 2000;404 (6776):394-8
[10]赵永波、陈耀明,帕金森病遗传学研究,上海交通大学学报医学版[J],2006,6(7)：708-710.
[11]张磊、张思瑾、徐晓恒,帕金森病的遗传研究进展,神经损伤与功能重建[J],2009,6(4)：444-449.
[12]Poiymeropoulos MH,Lavedan C,Leroy E, et al. Mutation in thea-synuclein gene identified in families with Parkinson's disease [J]. Science,1997,276(5321): 2045-2047.
[13]熊中奎、 胡雅儿,α—突触核蛋白的生物学功能及其在帕金森病中的作用,中国病理生理杂志[J],2010,26(9)：1855-1858.
[14]Kitada T,Asakawa S, Hattori N, et al. Mutation in the parkin gene cause autosomal recessive juvenile parkinsonism[J] Nature,1998,392(6676):605-608.
[15]ShermanMY,Goldberg AL. Cellular defenses against unfolded proteins:a cell biologist thinks about neurodegenerative diseases [J]. Neuron,2001,29 (1):15-32.
[16]陈鑫,牛平,泛素-蛋白酶体系统在帕金森病中的作用,解剖科学进展[J]2010,16(4)：370-373.
[17]Mcnaught KS,Olanow CW. Proteolytic stress:a unifying concept for the etiopathogenesis of Parkinson's disease[J]. AnnNeurol,2003,53 (Suppl 3):S73-84.
[18]王雪晶、郭纪锋、江泓帕金森病相关蛋白Parkin与PINK1的相互作用研究,生物化学与生物物理进展[J]2010,37(9)：983～987.
[19]Leroy E,Boyer R,Auburger G, et al. The ubiquitin pathway in Parkinson's disease[J]. Nature,1998,395(6701):451-457.
[20]沈海平,抗氧化应激对帕金森病的神经保护,现代中西医结合杂志,2010.19(6)：2096-2070
[21]REITER R J. Oxidative processes and ant ioxidat ive defense mechanisms in the aging brain[J]. FASEB J,1995,9(7):526.
[22]Junn E,Mouradian MM. Apoptotic signaling in dopamine-induced cell death:the role of oxidative stress,p38mitogen-activated protein kinase,cytochrome c and caspases[J]. J Neurochem,2001,78(2):374-383.
[23]Kost rzew a RM, Kost rzew a JP, Bru sR. Neu rop rotective and neu rotox ic roles of levodopa (L-DOPA) inneurodegenerative disord ers relat ing to Parkinsons d isease[J]. Am ino Acids,2002,23 (1/3):57-63
[24]Schapir a AHV, Copp er JM. Mi tochon drial comp 1I d ef iciency in Parkin son's di sease[J]. J Neu roch em,1990,54:823
[25]曹三勇、陈彪,泛素-蛋白酶体系统在帕金森病研究中的进展,中华神经科杂志[J],2009,39(4)：281-282
[26]贾晓晶,张秦玲,侯吉光,谷氨酸对小鼠胚胎中脑原代细胞培养中多巴胺能神经元损伤的时程效应,吉林大学学报医学版,2010.,36(6),1075-1078。
[27])宋明旭、杨吉成、王蔚平,谷氨酸诱导的PC12细胞损伤的研究,苏州科技学院学报自然科学版[J],2006.1(23)：62-65。
[28]费娜、 许丽珍,钙离子拮抗剂对帕金森病的神经保护作用,中国误诊学杂志[J],2006,6(7)：1244-1246。
[29]Langston JW. Epidemiology versus genetics in Parkinson's disease:progress in resolving an age-old debate [J]. Ann Neurol,1998,44 (3Suppl 1):S45-52.
[30]Iacopino AM, Christakos S. Specific reduction of calcium-binding protein (28-kilodalton calbindin-D) gene expression in aging and neurodegenerative diseases[J]. Proc Natl Acad Sci USA,1990,87 (11):4078-4082.
[31]Cho BP, Song DY, Sugam a S, et a.l Pa tho log ical dynam ics of ac tivated m ic rog lia fo llow ing m edial forebra in bund le transection [J]. G lia,2006,53(1):92-102
[32]张艳玲、苏炳银,帕金森发病免疫机制的研究进展,实用老年医学[J],2009,23(2) ： 150-152
[33]Tansey MG, Frank-Cannon TC, M c-Coy MK, e t a.l Neuro in flamma tion in Pa rkinsons disease is there sufficient ev idence for m echan ism based interventional therapy? [J]. Front B iosc,i 2008,13:709-717.
[34]Hunot, S, Hirsch, EC, Neuroinflammatory in Pakins'disease, Ann Neural,2003, 53 (3 supple 1):s49-s60。
[35]Mirza B, Hadberg H, Thom sen P, et al. The absence of reactive astrocytosis is indicative of a unique inflammatory process in Parkinson's disease. Neuroscience, 2000,95:425-432.
[36]Fiskum G, S tarkov A, Polster BM, et al. Mitochondrial mechanisms of neural cell death and neuroprotective interventions in Parkinson's disease. Ann N Y Acd Sci, 2003,991:111-119.
[37]安利佳、李志刚、姜波,帕金森病与细胞凋亡,中国现代医学杂志2006,16(7)：1032-1036
[38]Rao RV, Bredesen DE. Misfolded proreins, endoplasmic reticulum stress and neurodegeneration. CurrOpin in Cell.Biol,2004,16:653-662.
[39]M izuno Y, M oriH, Kondo T, Pract ical gu idelin es for the drug treatm ent of Park in son (?) sd isease, CNS Durgs 1994 1; 410-426.
[40]何得宾、尹寿玉,帕金森病的药物研究现状及进展,牡丹江医学院学报,2008,29(3) ： 88-90
[41]孟慧、孙静慧,帕金森病的药物治疗第四军医大学学报,2005,26(11),1053-1054
[42]Parkinson study Group. Evaluation of dyskinesias in a pilotrandomized, placebo controlled trial of remacemide in advanced Parkinson disease [J]. Arch Neurol,2001,58 (10):1 660～1 668
[43]林泉峰、李俊燕, 帕金森病治疗的进展,中国实用神经疾病杂志,2010.13(5)：88-90
[44]李勇杰,、 庄平、石长青等。 帕金森病患者丘脑底核的微电极定位技术[J].中华神经外科杂志,2005,21(1)：25-28.
[45]苏维海,帕金森病综合治疗的研究进展,实用心脑肺血管病杂志,2010,18(11)：1728-1730
[46]邵明,帕金森病的现代治疗与进展,神经损伤与功能重建,2007,2(6)：359-361)
[47]姚谦明,徐如祥,姜晓丹.转基因治疗帕金森病的研究进展.中华神经医学杂志,2006,5(2)：207-208.