六氯苯对PC-12细胞凋亡的影响

Effects of Hexachlorobenzene on the Apoptosis of PC-12 Cells

目的:探讨环境内分泌干扰物六氯苯对PC-12细胞凋亡的影响及其可能机制。方法:采用体外细胞培养方法,5%CO2 37℃恒温条件下,将PC-12细胞培养于含10%灭活胎牛血清的高糖DMEM培养基中。0、1、10、20、100、200μmol.L-1六氯苯处理组,每组设3组复孔,培养24 h后应用Cell Counting Kit-8试剂盒进行六氯苯对PC-12细胞增殖和毒性分析;流式细胞术FITC-An-nexinV/PI双染检测细胞凋亡率;Hoechst33258染色及倒置荧光显微镜拍摄细胞平片,观察细胞形态学改变;免疫印记法(Western blot)检测相关蛋白PLCγ及ERK蛋白磷酸化表达变化。结果:随着六氯苯浓度增加,细胞凋亡率升高,呈剂量依赖关系;Hoechst33258染色可见细胞核膨大、染色质边集浓染等凋亡特征;与对照组相比p-PLCγ及p-ERK1/2表达均降低,呈剂量依赖效用。结论:六氯苯能够诱导PC-12细胞凋亡,p-PLCγ及p-ERK1/2信号通路可能在此过程中发挥作用。

Objective：To investigate the effects of Hexachlorobenzene on the apoptosis of PC-12 cells.Methods：The PC-12 cells were cultured in high glucose Dulbecco＇s modified Eagle＇s medium（DMEM） and supplemented with 10 % fetal bovine serum,100 μg.ml-1 penicillin,and 100 μg.ml-1 streptomycin at 37℃ in a 5% CO2 atmosphere.Treated with Hexachlorobenzene（0,1,10,20,100,200 μmol.L-1） for 24 hours.Cell Counting Kit-8 was used to detected the proliferation and to do toxicity analysis.The morphological changes of PC-12 cells were observed by microscope,and cell apoptosis rates were detected by flow cytometry.The expression of p-PLCγ and p-ERK1/2 were determined by Western blot.Results：After treated with Hexachlorobenzene,PC-12 cells had morphologi-cal changes and higher apoptosis rates.The changes was evident with the increasing of the dose of Hexachlorobenzene.The 50% inhibit-ing concentration was 72 μmol.L-1.The apoptosis rate of 200 μmol.L-1 was 87.12 ± 3.21%.Conclusions：Hexachlorobenzene could in-duce apoptosis of PC-12 cells,and the expression of p-PLCγ and p-ERK1/2 proteins may play a role in the process of PC-12 cells apop-tosis induced by Hexachlorobenzene.