氟虫腈对PC12细胞凋亡及Bcl-2蛋白表达的影响

Effect of fipronil on apoptosis rate and Bcl-2 of PCI2 cells

目的 研究氟虫腈对神经细胞凋亡的影响并初步探讨其神经毒作用机制.方法 设3.13×10^-6、1.25×10^-5、5.00×10^-5mol/L 3个氟虫腈染毒组和1个对照组,用荧光显微镜检测各组PC12细胞凋亡荧光强度并用流式细胞仪对凋亡率进行测定,并对3个染毒组Bcl-2蛋白表达进行荧光强度检测.结果 细胞凋亡荧光强度检测发现,3.13×10×10^-6mol/L染毒组细胞凋亡率为17.00%±0.57%,5.00×10×10^-5mol/L染毒组细胞坏死率为13.95%±1.95%,均明显高于对照组（分别为9.80%±0.30%、3.10%±0.40%）,差异均有统计学意义（P〈0.05）.对Bcl-2蛋白表达含量进行定性检测,结果显示,3.13×10^-6mol/L染毒组Bcl-2蛋白表达明显低于对照组,其他染毒组Bcl-2蛋白表达与对照组比较无明显差异.结论 氟虫腈在较低剂量时可能是通过影响Bcl-2蛋白表达来诱导PCI2细胞凋亡,而在较高剂量时可能通过其他非细胞凋亡途径直接致细胞坏死发生.

Objecfive To study the apoptosis and mechanism of Fipmnil on PC12 Cells.Methods The effect of fipmnil on the apoptosis and necrosis of PCI2 cells of 3.13×10^-6, 1.25×10^-5,5.00×10^-5 mol/L three dose groups after 24 h treatment was detected by morphology and the apoptosis rate Was detected by flow cytometer（FCM）The expression of Bcl-2 protein in PC12 ceUs of 3.13×10^-6,1.25×10^-5）and 5.00×10^-5mol/L three dose groups after 24 h treatment Was measured by immunofluoreseence.Results The number of apop-totic cells of 3.13×10^-6mol/L group was more than the control group examined by fluorescence microscope,and the number of dead cells of 5.00×10^-5mol/L group was more than the control group.The apoptotic rates of Pc12 cells wag higher in the 3.13×10^-6mol/L group than the control group measured by FCM,and the dead rates of PC12 eels was higher in the 5.00×10^-5mol/L group than the control group（P〈0.05）.Immunonnores-cence eytochemistry experiment demonstrated that the level of Bcl-2 expression was significantly lower in the 3.13×10^-6mol/L group than the control group.Conclusion At low dosage,fipronil increases the apoptotic rates of PC12 cells possibly by decreasing the expression of Bcl-2 protein while at high dosage,fipmnil only in-creases the amount of necmtic cells.