IL-1β在溴氰菊酯诱发谷氨酸释放中的作用

Function of interleukin 1β in the glutaminic acid releasing from neural unicells exposed to DM

目的探讨IL-1β与Glu在溴氰菊酯(DM)神经毒性中的关系。方法分离雄性SD大鼠皮层、海马和下丘脑,消化成单细胞悬液置37℃培养,3个脑区的细胞均分为对照组:给予体积分数为0.1%DMSO;DM组:给予2×10-6mol/L DM;DM1组:给予2×10-6mol/L DM同时加10μmol/L IL-1β转化酶抑制剂Ac-YVAD-CHO;DM2组:给予2×10-6mol/L DM后15 min加10μmol/L Ac-YVAD-CHO;DM3组:给予2×10-6mol/L DM同时加15μmol/L Ac-YVAD-CHO;DM4组:给予2×10-6mol/L DM后15 min加15μmol/L Ac-YVAD-CHO,每组3个平行样,DM作用1 h后收集培养上清,高效液相色谱(HPLC)法测上清中Glu释放量。结果DM可显著促进皮层和海马神经细胞的Glu释放,但抑制下丘脑细胞Glu释放;皮层和海马各抑制剂组Glu的释放均显著低于DM组但仍高于对照组,DM作用后15 min给予10μmol/L,Ac-YVAD-CHO对皮层细胞的Glu释放的抑制作用较先给予同等剂量的抑制剂作用强(P<0.01);对海马神经细胞,15μmol/L抑制剂对DM诱导Glu释放的抑制作用较10μmol/L抑制剂作用强,DM作用后15 min再给予15μmol/L较先给予15μmol/L抑制剂的抑制作用强;而Ac-YVAD-CHO对DM诱导下丘脑细胞谷氨酸(Glu)释放的抑制作用各抑制组间无差别。结论IL-1β促进了DM对皮层和海马神经细胞Glu的诱导作用。

Objective To explore the relationship between IL-1β and glutaminic acid （Glu） involved in the neurotoxieity of dehamethrin. Methods Cortex, hippocampus and hypothalamus were isolated from male SD rats and were digested under 37 ℃ for unicell suspension, then the cell suspensions of three cerebral regions were separated into 6 groups respectively ：eontrol group （V ： V = 0.1% DMSO）, DM group （2 × 10^-6 mol/L DM）, DM1 group （added 10 μmol/L IL-1β converting enzyme inhibitor Ac-YVAD-CHO and 2 × 10^-6 mol/L DM simultaneously）, DM2 group （added 10 μmol/L Ac-YVAD-CHO 15 min after DM treatment）, DM3 group （added 15 /μmol/L Ac-YVAD-CHO and 2 × 10^-6 mol/L DM simultaneously）, DM4 group （added 15 μmol/L Ac-YVAD-CHO 15 min after DM treatment）, each group contained 3 parallel samples. 1 h after DM treatment, supernatants were collected to detected the GLU by HPLC. Results DM promoted GLU releasing from cortical and hippocampal cells but not hypothalamic cells; Ae-YVAD-CHO could reduced GLU releasing of cortex and hippoeampus dramatiely compared with the two DM groups; 10 μmol/L Ac-YVAD-CHO 15 min after DM treatment in DM2 group inhibited GLU releasing more efficiently than DM1 group of cortical cells; in hippocampal cell groups, 15 μmol/L Ac-YVAD-CHO inhibited GLU releasing more efficiently than 10μmol/L Ae-YVAD-CHO, and the DM4 group inhibited more efficiently than the DM3 group. Conclusions IL-1β involved in GLU releasing of cortical and hippocampal cells exposed to DM.