α-突触核蛋白的表达与氧化应激水平的相互影响

摘要

Objective To investigate the interaction between the oxidative stress and the expression of α- synuclein protein in PC12 cells. Methods The levels of reactive oxygen species (ROS) in various groups of PC12 cells, including non-transfected, transfected with empty vector, transfected with wild-type α- synuclein and transfected with mutant-type α- 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 α-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＜0.05). 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 α-synuclein in the WT-type and A53T-type PC12 cells.%目的 探讨氧化应激与在稳定转染α-突触核蛋白(α-synuelein)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 测定末转染、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PCI2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、1000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.