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目的探讨Fas/Fas L信号在铅诱导大鼠肾上腺嗜铬细胞瘤细胞(PC12细胞)发生细胞凋亡中的作用。方法醋酸铅的暴露剂量设为0、0.1、1.0与10.0μmol/L,暴露时间为48 h。免疫荧光技术观察细胞内Fas蛋白的表达,Westernblot法检测Fas与Fas L蛋白表达水平,比色法测定细胞内caspase-8与caspase-3的活性,流式细胞术检测细胞凋亡率。结果免疫荧光技术与Western blot法均表明,与对照组比较,1.0与10.0μmol/L醋酸铅暴露可致PC12细胞内Fas表达明显增加(P<0.05),10.0μmol/L醋酸铅可诱导Fas L蛋白表达显著增加(P<0.01)。此外,与对照组相比,caspase-8活性在10.0μmol/L醋酸铅暴露后显著升高(P<0.01),而caspase-3活性在1.0与10.0μmol/L醋酸铅染毒后均显著上升(P<0.05)。流式细胞术结果显示,0.1、1.0与10.0μmol/L醋酸铅染毒均增加PC12细胞凋亡率(P<0.05),与对照组比较,差异有统计学意义。结论铅暴露可诱导PC12细胞凋亡,其机制可能与死亡受体途径的激活有关。
Objective To investigate the role of Fas / Fas L signaling in the apoptosis of lead-induced adrenal pheochromocytoma cells (PC12 cells). Methods Exposure doses of lead acetate were set at 0, 0.1, 1.0 and 10.0 μmol / L for 48 h. The expression of Fas protein in cells was observed by immunofluorescence. The expression of Fas and Fas L protein was detected by Western blotting. The activity of caspase-8 and caspase-3 was detected by colorimetric assay. The apoptosis rate was detected by flow cytometry. Results Both immunofluorescence and Western blot showed that the expression of Fas protein in PC12 cells was significantly increased (P <0.05) when exposed to 1.0 and 10.0 μmol / L lead acetate compared with the control group, and 10.0 μmol / L lead acetate induced Fas L protein expression Significantly increased (P <0.01). In addition, caspase-8 activity was significantly increased (P <0.01) after exposure to 10.0 μmol / L lead acetate compared to the control group, while caspase-3 activity was significantly increased after exposure to 1.0 and 10.0 μmol / L lead acetate <0.05). Flow cytometry results showed that 0.1,1.0 and 10.0μmol / L lead acetate exposure both increased the apoptosis rate of PC12 cells (P <0.05), compared with the control group, the difference was statistically significant. Conclusion Lead exposure can induce the apoptosis of PC12 cells, which may be related to the activation of death receptor pathway.