摘要
目的探讨铅暴露对星形胶质细胞（AS）中GLAST和GLT-1蛋白表达及转运体功能的影响。方法取出生1-3dWistar大鼠的仔鼠星形胶质细胞进行原代培养，以胶质纤维酸性蛋白（GFAP）染色鉴定细胞纯度后，进行0（对照组）、50、100、200及400μmol／L乙酸铅染毒，染毒时间为72h。倒置显微镜下观察细胞形态并采集图像；以AlamarBlue法检测细胞活力；以Westernblot法检测GLAST和GLT-1蛋白的表达；以同位素标记法测定谷氨酸转运体功能。结果50和100μmol／L铅暴露组的细胞形态与对照组相比，未见明显改变；200μmol／L铅暴露72h后，少量细胞开始脱壁，细胞间隙增大；400μmol／L铅暴露组细胞变大，突起变短或消失，脱壁细胞数量明显增多。与对照组相比，各浓度铅暴露组的AS活力均降低（P〈0．05），100、200和400μmol／L铅暴露组的GLT-1蛋白含量下降（P〈0．05），但各浓度铅暴露组的GLAST蛋白含量无显著改变。100、200和400μmol／L铅暴露组的3FI—Glu放射性低于对照组，差异均有统计学意义（P〈0．05）。结论铅暴露可明显抑制AS中GLT-1蛋白的表达，并导致AS的Glu转运体功能下降。
Objective To investigate the impact of lead exposure on the protein expression of GLAST and GLT-1 and the function of glutamate transporters in primary cultured astrocytes of rats. Methods The astrocytes were separated by mechanical method from Wistar rats aged one to three days and immunocytochemically stained with anti-GFAP antibody to identify. The cultured cells were randomly divided into five groups, in which astrocytes were exposed to 0, 50, 100, 200 and 400 μmol/L PbAc2 respectively for 72 h. Phase contrast microscope was used to observe the morphological changes. Alamar blue was used to assess the effects of PbAc2 on the cell proliferation capacity. Protein expression level of GLAST and GLT-1 were detected with Western blot. 3H radiolabeled method was used to determinate the function of glutamate transporter. Results Compared with the control group, in 50 and 100 μmol/L lead exposure groups, the cells had no significant morphological change. In 200 μmol/L lead exposure group, a small number of cells fell off the wall and the cell-gap increased. In 400 p, mol/L lead exposure group, the cells became larger, protrusion was shorter or disappeared. The cell viability of all group were apparently declined with the increasing dose of lead compared with that in control group. The contents of GLT-1 protein in 100, 200 and 400 μmol/L lead exposure groups were also remarkably diminished. However, there were no significant changes in GLAST protein content of all groups. The uptake of radioactive glutamate in 100, 200 and 400μmol/L lead exposure groups revealed a significant reduction compared with the control group. Conclusion Lead exposure may inhibit the GLT-1 protein expression and decrease the activity of Glu transporter in astroeytes.
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2014年第3期193-196,F0003,共5页
Journal of Environment and Health
基金
国家自然科学基金(31070992)
辽宁省自然科学基金(20102263
201102276)
辽宁省教育厅科研基金项目(L2010559)
