预热激对6-OHDA诱导SH-SY5Y细胞神经毒性损伤的保护作用

Neuroprotective effect of pre-heat shock on 6-OHDA-induced neurotoxicity in SH-SY5Y cells

目的探讨预热激对6-羟多巴胺(6-OHDA)诱导人神经母细胞瘤细胞(SH-SY5Y)神经毒性损伤的保护作用及其机制。方法对实验组SH-SY5Y细胞进行预热激(42±0.5)℃处理,再通过6-OHDA对其进行神经毒性诱导,未行预热激处理细胞作为对照组。逆转录酶-聚合酶链式反应(RT-PCR)检测不同热激时间(15 min、30 min、60 min)SH-SY5Y细胞内葡萄糖调节蛋白(GRP78、GRP94)的表达水平;显微镜下观察细胞形态学变化,并用四甲基偶氮唑蓝(MTT)比色法检测SH-SY5Y细胞活性情况。结果 6-OHDA对SH-SY5Y细胞的神经毒性损伤呈剂量依赖性及时间依赖性关系,实验组SH-SY5Y细胞较对照组所受的神经毒性损伤轻(P<0.05),且实验组间差异也具有统计学意义(P<0.05);SH-SY5Y细胞中GRP78和GRP94 mRNA表达水平与热应激时间正相关(P<0.05)。结论预热激可降低6-OHDA诱导SH-SY5Y细胞的神经毒性损伤作用,且热激过程中SH-SY5Y细胞中GRP78和GRP94表达上调。

Objective To investigate the neuroprotective effect and mechanism of pre-heat shock treatment on 6-hydroxydopamine（6-OHDA）-induced neurotoxicity in SH-SY5Y cells.Methods Following the pre-heat shock at 42℃±0.5℃,6-OHDA-induced neurotoxicity was conducted in experimental group.In control group only 6-OHDA-induced neurotoxicity was carried out.Expressions of glucose regulated protein（GRP78 and GRP94） in SH-SY5Y cells were detected at different time points（15 min,30 min,60 min） by reverse transcription-polymerase chain reaction（RT-PCR）.The morphological changes of the cells were observed under microscope and the cell viability was identified by MTT（3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） assay.Results 6-OHDA induced the neurotoxicity of SH-SY5Y cells in a dose-and time-dependent manner.6-OHDA-induced neurotoxicity on SH-SY5Y cells was less in experiment group compared with that in control group and the statistic difference among different time points in experimental group was significant（P0.05）.The mRNA levels of GRP78 and GRP94 showed a time-dependent relationship with the extension of pre-heat shock time（P0.05）.Conclusion Pre-heat shock treatment can reduce 6-OHDA-induced neurotoxicity of SH-SY5Y cells and the expressions of GRP78 and GRP94 are increased during the heat shock treatment.