α-突触核蛋白的表达与氧化应激水平的相互影响  

The interaction between the oxidative stress level and α-synuclein protein expression

作　　者：沈原[1] 赵永波[1] 刘功禄[1] 赵圣杰[1] 

机构地区：[1]上海交通大学附属第一人民医院神经科,200080

出　　处：《中国神经免疫学和神经病学杂志》2011年第3期174-177,共4页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家自然科学基金资助项目(30270493);上海市引进海外高层次留学人员专项基金资助项目(20040115);上海自然科学基金资助项目(01ZB14050);上海市科学技术发展基金重点资助项目(024119037);上海交通大学优秀中青年科研基金资助项目(2005-3-8)

摘　　要：目的探讨氧化应激与在稳定转染α-突触核蛋白(-αsynuclein)的大鼠嗜铬细胞瘤细胞(PC12细胞)中α-突触核蛋白表达的相互影响。方法测定未转染、转染空质粒、已稳定转染野生型(WT)及突变型(A53T)α-突触核蛋白的PC12细胞的活性氧(ROS)水平,并以实时定量PCR法和免疫印迹(Western blot)法检测分析经0、50、100和250 nmol/L H2O2处理的WT型和A53T型PC12细胞α-突触核蛋白表达。结果 (1)α-突触核蛋白转染PC12细胞后,细胞内ROS水平增加,按未转染的PC12细胞、转染空质粒的PC12细胞、WT型PC12细胞、A53T型PC12细胞顺序ROS水平依次增加(P<0.05)。(2)经不同浓度H2O2处理后,WT型和A53T型PC12细胞α-突触核蛋白mRNA表达水平均无明显改变,WT型及A53T型PC12细胞不同浓度组组间差异均无统计学意义(均P>0.05);在Western blot检测中,50、100和250 nmol/L H2O2组WT型PC12细胞α-突触核蛋白表达较0 nmol/L H2O2组减少,且不同浓度组组间差异均有统计学意义(P<0.05);50、100和250nmol/L H2O2组A53T型PC12细胞α-突触核蛋白表达较0 nmol/L H2O2组增加,且不同浓度组两两比较差异亦均有统计学意义(P<0.05)。结论α-突触核蛋白可引起PC12细胞氧化应激损伤,且不同氧化应激水平对WT型和A53T型PC12细胞中的α-突触核蛋白表达有不同程度的影响。Objective To investigate the interaction between the oxidative stress and the expression of α- synuelein protein in PC12 cells. Methods The levels of reactive oxygen species （ROS） in various groups of PC12 cells, including nomtransfected, transfected with empty vector, transfected with wild-type α- synuclein and transfected with mutant-type m synuclein, were detected. The expression of α- synuclein was analyzed by realtime PCR and Western blot in the WT-type and A53T-type PC12 cells after the treatment of different concentrations of H2O2 （0, 50, 100 and 250 nmol/L）. Results （1） α- synuclein increased the levels of intracellular ROS. Based on the increased amount of intracellular ROS, the PC12 cell groups were lined up as following： PC12 cells 〈 empty-vector-type PC12 cells 〈 WT-type PC12 cells 〈 A53T type PC12 cells （P〈 0.05）. （2） Different concentrations of H2O2 did not cause statistical significance in the mRNA expression of α- synuclein in the WT-type PC12 cells （P〉0.05）, neither in the A53T-type PC12 cells （P〉0.05）. In Western blotting test, the expression of a synuclein in the WT-type PC12 cells decreased in 50, 100 and 250 nmol / L H2O2 group as compared with 0 nmol / L H2O2 concentration group, and significant difference was also present among the different H2O2 concentration groups （P〈C0.05）. The expression of α-synuclein in the A53T-type PC12 cells increased in 50, 100 and 250 nmol / L H2O2 group as compared with 0 nmol / L H2O2 concentration group, and the increase was significant among the different concentration groups （P〈0.05）. Conclusions α- synuclein caused oxidative stress on the PC12 cells, and the different levels of oxidative stress had different influences on the expression of a synuclein in the WT-type and A53T type PC12 cells.

关 键 词：Α-突触核蛋白 活性氧 氧化应激 PC12细胞 

分 类 号：R742.5[医药卫生—神经病学与精神病学]