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 (P ＜0.05). The mRNA levels of GRP78 and GRP94 showed a time-dependent relationship with the extension of preheat shock time (P ＜0.05). 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.%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.
展开▼
