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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、200 μmol.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-AnnexinV/ 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。

英文摘要:

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 morphological changes and higher apoptosis rates. The changes was evident with the increasing of the dose of Hexachlorobenzene. The 50% inhibiting concentration was 72 μmol.L-1. The apoptosis rate of 200 μmol.L-1 was 87.12 ± 3.21%. Conclusions: Hexachlorobenzene could induce 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 apoptosis induced by Hexachlorobenzene.

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