应激和分子伴侣对鱼藤酮诱导的SH-SY5Y细胞毒性的保护作用

Protective effect of STCH on rotenone induced neurotoxicity of human SH-SY5Y cells

目的:本文探讨应激和分子伴侣(stress and chaperone,STCH)对鱼藤酮的神经毒性之于SH-SY5Y细胞的保护作用及其可能的作用机制。方法:采用不同浓度鱼藤酮处理SH-SY5Y-pcDNA 3.1和SH-SY5Y-pcDNA3.1-STCH稳定细胞株,并观察细胞形态;采用MTT法和Western蛋白印迹法检测MAPK通路相关的信号分子和凋亡相关蛋白变化。结果:与未处理组相比,鱼藤酮均能引起细胞活力显著下降(P<0.05);STCH对鱼藤酮诱导的SH-SY5Y细胞毒性具有明显的保护作用(0.1μmol/L-5μmol/L),但10μmol/L浓度组与转染空载体组无显著差异。激活型caspase-3蛋白在鱼藤酮毒素处理后表达明显,p38磷酸化增强,过表达STCH在鱼藤酮处理后p38磷酸化受到明显抑制,裂解的caspase-3活性形式减少。结论:STCH对鱼藤酮诱导神经毒性的保护作用可能通过抑制p38磷酸化而实现。

Objective： We explored the protective effects and possible mechanism of stress and chaperone （STCH） on rotenone-induced neurotoxicity in SH-SYSY. Methods： Human SH-SY5Y-pcDNA3.1 and SH-SYSY-pcDNA3.1-STCH cells were treated with rotenone of different concentration, respectively; and the cell morphology was observed. The cell viability was evaluated by MTr assay and molecules related to MAPK signaling pathway and cell apoptosis were examined by Western Blot. Results： Our data showed that rotenone induced a decrease of cell viability in SH-SYSY （P 〈0.05） compared to control. Compared to empty vector transfected group, STCH showed a protective role with concentrations ran- ging from 0.1 μmol/L to 5 μmol/L of rotenone （P 〈0.05） while there was no significant difference at 10 μmol/L of ro- tenone as demonstrated by SH-SYSY-pcDNA3.1 and SH-SYSY-pcDNA3.1-STCH cells groups. Compared to non-treat- ment group, increase of cleaved caspase-3 form and phosphorylation of p38 after treatment with rotenone in SH-SYSY- pcDNA3.1 were detected respectively. While in SI-I-SYSY-pcDNA3.1-STCH group （ over-expression of STCH following rotenone treatment）, activation of cleavage of caspase-3 form and phosphorylation of p38 were suppressed. Conclusion： These data indicate that the protective effects of STCH against rotenone-induced neurotoxicity may be mediated by sup- pressing activation of the p38 signaling-induced apoptosis.