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目的探讨存活素(Survivin)能否抑制(1-甲基-4-苯基-吡啶离子(1-methyl-4-phenylpyridinium,MPP~+)诱导的神经元(SH-SY5Y细胞)的凋亡过程。方法运用1 mmol/L MPP~+处理SH-SY5Y细胞,诱导其发生凋亡,建立帕金森病的细胞模型,采用不同浓度的Survivin进行预处理,使用倒置显微镜观察细胞形态,采用MTT法、Real-time PCR、Western blot检测相关指标,进而评价Survivin对SH-SY5Y凋亡的作用。结果 1 mmol/L MPP~+作用24 h可诱导SH-SY5Y的凋亡,显著降低细胞存活率(F=13.32,P=0.000)。MPP~+作用前,预先经Survivin处理2 h的SH-SY5Y细胞的存活率显著提高(F=13.32,P=0.000,P=0.000,P=0.000),并呈剂量依赖性关系。Real-time PCR结果显示Survivin能够显著抑制SH-SY5Y细胞中凋亡蛋白Caspase 3和Caspase 9的表达水平(F=61.57,P=0.000,P=0.000,P=0.000;F=54.26,P=0.001,P=0.000,P=0.000);Western blot结果亦显示Survivin能够显著抑制SH-SY5Y细胞中凋亡蛋白Caspase 3和Caspase 9的表达水平。结论 Survivin能够剂量依赖性地抑制MPP~+诱导的SH-SY5Y细胞凋亡,其机制可能与阻断Caspase信号通路相关。
Objective To investigate whether Survivin can inhibit the apoptosis of neurons (SH-SY5Y cells) induced by 1-methyl-4-phenylpyridinium (MPP ~ +) .Methods SH-SY5Y cells were treated with 1 mmol / L MPP ~ + to induce apoptosis. Cell model of Parkinson’s disease was established. Survivin was used to pretreat SH-SY5Y cells. The morphological changes of the cells were observed by MTT method. Real-time PCR and Western blot were used to evaluate the effect of Survivin on the apoptosis of SH-SY5Y cells.Results The apoptosis of SH-SY5Y cells induced by 1 mmol / L MPP ~ + for 24 h significantly decreased the cell viability (F = 13.32, P = 0.000). The survival rate of SH-SY5Y cells pretreated with Survivin for 2 h was significantly increased (P = 0.000, P = 0.000, P = 0.000) The results of Real-time PCR showed that Survivin could significantly inhibit the expression of Caspase 3 and Caspase 9 in SH-SY5Y cells (F = 61.57, P = 0.000, P = 0.000, P = 0.000; 54.26, P = 0.001, P = 0.000, P = 0.000). Western blot also showed that Survivin could significantly inhibit the expression of Caspase 3 and Caspase 9 in SH-SY5Y cells Level. Conclusion Survivin can be dose-dependently inhibited MPP ~ induced apoptosis of SH-SY5Y cells +, which may be related to the blocking Caspase signal pathway.