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目的研究三邻甲苯磷酸酯(TOCP)对人成神经瘤细胞(SH-SY5Y)calpain活性的影响。方法用全反式维甲酸诱导SH-SY5Y细胞分化,用不同浓度的TOCP(0、0.25、0.5、1.0mM)染毒未分化或分化的SH-SY5Y细胞不同时间(12、24、48h),用20μM盐酸维拉帕米(verapamil)和100μM二苯基硼酸-2-氨基乙酯(2-APB)预处理细胞15min后染毒TOCP 1.0mM 48h,采用荧光分光光度法测calpain活性。结果随着染毒浓度和时间的增加,TOCP活化未分化与分化的SH-SY5Y细胞calpain活性(P<0.05),钙通道抑制剂verapamil和2-APB预处理细胞能抑制TOCP引起的calpain活性升高(P<0.05)。结论 TOCP通过SH-SY5Y细胞内钙紊乱而活化calpain活性。
Objective To investigate the effect of tri-o-tolyl phosphate (TOCP) on calpain activity in human neuroblastoma cells (SH-SY5Y). Methods SH-SY5Y cells were induced by all-trans retinoic acid. Undifferentiated or differentiated SH-SY5Y cells were treated with different concentrations of TOCP (0,0.25,0.5,1.0mM) for different times (12,24,48h) The cells were pre-treated with 20μM verapamil and 100μM 2- (2-APB) for 15min and then TOCP 1.0mM 48h. The calpain activity was measured by fluorescence spectrophotometry. Results TOCP activated the calpain activity of SH-SY5Y cells (P <0.05) with the increase of exposure concentration and time. Pretreatment with calcium channel inhibitor verapamil and 2-APB inhibited the increase of calpain activity induced by TOCP High (P <0.05). Conclusion TOCP activates calpain activity through an intracellular calcium disorder in SH-SY5Y cells.