三邻甲苯磷酸酯对人成神经瘤SH-SY5Y细胞calpain活性的影响

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(P0.05),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(P0.05)。结论 TOCP通过SH-SY5Y细胞内钙紊乱而活化calpain活性。

Effect of tri-o-cresyl phosphate on calpain activity in human neuroblastoma SH-SY5Y cells

ObJective To study the impact of tri -O- cresyl phosphate （TOCP） on the activity of calpain in human neuroblas- toma SH- SY5Y cells. Methods Retinoic acid was used to induce differentiation of SH- SY5Y cells. The undifferentiated or differentiated SH - SY5Y cells were treated with TOCP at different concentratiorks （0, 0.25, 0, 5 and 1,0 mM） for 12, 24 or 48 hours. In another experiment subset, the cells were pretreated with 20 μM verapamil, 100 μM 2 - aminoethoxydiphenyl bo- rate （2 - APB） or both for 15 minutes and then treated with 1.0 mM TOCP for 48 hours. The activity of calpain was detected by fluoroscence spectrophotometry. Results The activity of calpain in undifferentiated and differentiated SH - SY5Y cells were increased gradually as the dose and time increased （P〈 0.05）. TOCP- induced calpain activity increments in SH- SY5Y cells could be significantly inhibited by pretreatment with calcium channel blockers, verapamil and 2 - APB （P 〈 0.05）. Con- elusiotl, s TOCP can increase the activity of calpain in SH- SY5Y cells through intracellular calcium disorder.