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突变型α-核突触蛋白的自噬性降解途径及可能机制 被引量:7

Autophagic pathway and probable mechanism in degradation of mutant α-synuclein in PC12 cells
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摘要 目的观察突变型α-核突触蛋白对PCI2细胞增殖的影响和可能的降解途径,探讨其在帕金森病发病机制中的作用。方法对转染了α-核突触蛋白(A30P)的PCI2细胞进行药物干预,检测细胞的增殖活性,并采用透射电镜观察细胞超微结构改变以及自噬的特征性改变,同时检测α-核突触蛋白的表达和超氧化物歧化酶(SOD)的水平。结果(1)Western Blot法检测α-核突触蛋白的表达:A30P+渥曼青霉素组(A30P+W组)、A30P+1-甲基-±苯基吡啶组(A30P+MPP^+组)较A30P组明显增高,以A30P+W组最为明显;而A30P+雷帕霉素组(A30P+R组)条带较A30P组减低(P〈0.01);(2)不同时间点细胞培养液中SOD水平(U/ml)的测定:用MPP^+处理转染了突变型α-核突触蛋白的PC12细胞后,培养液中SOD水平(A30P+MPP^+组:3h:97.49±13.8;12 h:102.7±12.7;24 h:101.5±11.8;48 h:104.3±12.4)较A30P组在各时间点显著下调(t=3.7721,P=0.0017);A30P+R组在给药12h以后,培养液中SOD水平逐渐升高,其中在24h(121.2±13.0)、48h(124.3±14.1)和72h(127.7±13.7)时与A30P+W组比较差异有统计学意义(t=2.9746,P=0.0083);突变型α-核突触蛋白激活了自噬途径,并介导了MPP^+的毒性作用,自噬抑制剂渥曼青霉素可通过抑制自噬而加剧α-核突触蛋白积聚,导致细胞死亡;而自噬诱导剂雷帕霉素则可以通过诱导自噬的发生而促进α-核突触蛋白的降解和细胞生长。结论α-核突触蛋白的异常积聚导致PC12细胞的自噬性细胞死亡,促进自噬有助于突变型α-核突触蛋白降解,对细胞具有保护作用。 Objective To observe the effect of mutant α-synuclein (A30P) in autophagic programmed cell death by transfected PC12 cells and explore its probable role and pathway in PD. Methods The definite PC12 cells which were transfected mutant α-synuclein (A30P) were constructed at first and MPP^+, Rapamycin and Wortmanin were administrated to transfected PC12 cells with mutant α-synuclein. Not only the proliferative activity of cells was detected with MTF method but also the uhrastructure changes of cells and expression of α-synuclein in different circumstance were observed by transmission electron microscopy (TEM), Western Blot and the level of SOD. Results ( 1 ) The expression of α-synuclein in groups A30P +Wortmannin and A30P +MPP^+was higher than that in group A30P ( P 〈 0. 01 ), particularly, there was more significant expression of α-synuclein in group A30P +Wortmannin. The expression of α-synuclein in group A30P +Rapamycin was weaker than that in group A30P ( P 〈 0. 01 ) ; (2) The results showed that the SOD level ( group A30P +MPP^+:3 h: 97.49 ± 13.8; 12 h: 102. 7 ± 12.7 ; 24 h: 101.5 ± 11.8; 48 h: 104. 3 ± 12. 4) was significantly decreased at various time points after MPP^+treatment compared that of group A30P (t =3. 7721, P =0. 0017). SOD level gradually increased in A30P + Rapamycin 12 h and showed significant difference at 24 h ( 121.2 ± 13.0), 48 h ( 124. 3 ± 14. 1 ) and 72 h (127. 7 ± 13. 7) after drug treatment compared with that in group A30P + Wortmannin (t = 2. 9746, P =0. 0083) ; (3) Mutant α-synuclein (A30P) leading to PC12 cells death by means of autophagy involved α-synuclein accumulation, membrane lipid oxidation, and loss of plasma membrane integrity. Mutant α- synuclein (A30P) mediated the toxicity of MPP^+. Rapamycin, an inducer of autophagy, reduced the aggregation of α-synuclein in transfected cells. Meanwhile, Wortmanin, an inhibitor of autophagy, promoted the aggregation of α-synuclein in transfected cells and induced cells to die. Conclusions The abnormal aggregation of α-synuclein induces autophagic programmed cell death in PC12 ceils and mutant α-synuclein (A30P) mediates the toxicity of MPP^+ Meanwhile, Rapamycin may reduce the aggregation of α-synuclein in transfected cells by activation of autophagic pathway.
出处 《中华神经科杂志》 CAS CSCD 北大核心 2008年第1期51-55,共5页 Chinese Journal of Neurology
基金 卫生部科学研究基金资助项目(WKJ2005-2-030)
关键词 帕金森病 PC12细胞 自吞噬作用 α-核突触蛋白 Parkinson disease PC12 cells Autophagocytosis α-synuclein
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参考文献29

  • 1Terman A, Gustafsson B, Brunk UT. Autophagy, organelles and ageing. J Pathol, 2007, 211: 134-143.
  • 2Kuma A, Hatano M, Matsui M, et al. The role of autophagy during the early neonatal starvation period. Nature, 2004, 432: 1032-1036.
  • 3Anglade P, Vyas S, Javoy-Agid F, et al. Apoptosis and autophagy in nigral neurons of patients with Parkinson' s disease. Histol Histopathol, 1997, 12: 25-31.
  • 4Rubinsztein DC. The roles of intracellular protein-degradation pathways in neurodegeneration. Nature, 2006, 443: 780-786.
  • 5Stefanis L, Larsen KE, Rideout HJ, et al. Expression of A53T mutant but not wild-type alpha-synuclein in PC12 cells induces alterations of the ubiquitin-dependent degradation system, loss of dopamine release, and autophagic cell death. J Neurosci, 2001, 21 : 9549-9560.
  • 6Spillantini MG, Schmidt ML, Lee VM, et al. Alpha-synuclein in Lewy bodies. Nature, 1997, 388: 839-840.
  • 7Masliah E, Rockenstein E, Veinbergs I, et al. Dopaminergic loss and inclusion body formation in alpha-synuclein mice: implications for neurodegenerative disorders. Science, 2000, 287: 1265-1269.
  • 8Giasson BI, Duda JE, Murray IV, et al. Oxidative damage linked to neurodegeneration by selective alpha-synuclein nitration in synucleinopathy lesions. Science, 2000, 290 : 985-989.
  • 9钱进军,程言博,刘春风,胡伟东.人SNCA及其致病突变基因的逆转录病毒pEGZ/MCSHA载体的构建[J].中国临床神经科学,2006,14(6):561-565. 被引量:3
  • 10Kruger R, Kuhn W, MUller T, et aL Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson' s disease. Nat Genet, 1998, 18: 106-108.

二级参考文献15

  • 1Polymeropoulos MH,Lavedan C,Leroy E,et al.Mutation in the alpha-synuclein gene identified in families with Parkinson's disease[J].Science,1997,276:2045-2047
  • 2Kruger R,Kuhn W,Muller T,et al.Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson's disease[J].Nat Genet,1998,18:106-108
  • 3Clayton DF,George JM.The synucleins:a family of proteins involved in synaptic function,plasticity,neurodegeneration and disease[J].Trends Neurosci,1998,21:249-254
  • 4Murphy DD,Rueter SM,Trojanowski JQ,et al.Synucleins are developmentally expressed,and alpha-synuclein regulates the size of the presynaptic vesicular pool in primary hippocampal neurons[J].J Neurosci,2000,20:3214-3220
  • 5Fortin DL,Troyer MD,Nakamura K,et al.Lipid rafts mediate the synaptic localization of a-synuclein[J].J Neurosci,2004,24:6715-6723
  • 6Perez RG,Waymire JC,Lin E,et al.A role for α-synuclein in the regulation of dopamine biosynthesis[J].J.Neurosci,2002,22:3090-3099
  • 7Fujiwara H,Hasegawa M,Dohmae N,et al.α-Synuclein is phosphorylated in synucleinopathies[J].Nat Cell Biol,2002,4:160-164
  • 8Masliah E,Rockenstein E,Veinbergs I,et al.Dopaminergic loss and inclusion body formation in alpha-synuclein mice:implications for neurodegenerative disorders[J].Science,2000,287:1265-1269
  • 9Hsu LJ,Sagara Y,Arroyo A,et al.alpha-synuclein promotes mitochondrial deficit and oxidative stress[J].Am J Pathol,2000,157:401-410
  • 10Lotharius J,Barg S,Wiekop P,et al.Effect of mutant α-synuclein on dopamine homeostasis in a new human mesencephalic line[J].J.Biol.Chem,2002,277:38884-38894

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同被引文献103

  • 1洪雁,张本恕,马温良.Parkin基因和帕金森病[J].国外医学(神经病学.神经外科学分册),2004,31(3):283-286. 被引量:2
  • 2Ya-ping YANG,Zhong-qin LIANG,Zhen-lun GU,Zheng-hong QIN.Molecular mechanism and regulation of autophagy[J].Acta Pharmacologica Sinica,2005,26(12):1421-1434. 被引量:51
  • 3张振涛,曹学兵,孙圣刚.帕金森病多巴胺神经元死亡的分子机制[J].中国康复,2006,21(6):402-404. 被引量:17
  • 4何延波,陈彪.α-synuclein基因多态性与帕金森病[J].中华神经科杂志,2007,40(2):133-135. 被引量:5
  • 5Klionsky DJ, Abeliovich H, Agostinis P, et al. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy,2008,4 : 151 - 175.
  • 6Mizushima N, Klionsky DJ. Protein turnover via autophagy: implications for metabolism. Annu Rev Nutr, 2007,27 : 19 - 40.
  • 7Levine B, Kroemer G. Autophagy in the pathogenesis of disease. Cell,2008,132 : 27 -42.
  • 8Martinez-vicente M, Cuervo AM. Autophagy and neurode- generation: when the cleaning crew goes on strike. Lancet Neurol,2007,6 : 352 - 361.
  • 9Rubinsztein DC, Gestwicki JE, Murphy LO, et al. Potential therapeutic applications of autophagy. Nat Rev Drug Discov, 2007,6 : 304 -312.
  • 10Bredesen DE. Programmed cell death mechanisms in neurological disease. Curr Mol Med,2008,8 : 173 - 186.

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