Hu Xiong-Fei, Li Jing, Zhuang Wan, Li Long
Department of Health Toxicology and MOE Key Lab of Environment and Health, School of Public Health, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2010 Apr;28(4):264-7.
To study the apoptosis and mechanism of Fipronil on PC12 Cells.
The effect of fipronil on the apoptosis and necrosis of PC12 cells of 3.13 x 10(-6), 1.25 x 10(-5) and 5.00 x 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 cells of 3.13 x 10(-6), 1.25 x 10(-5) and 5.00 x 10(-5) mol/L three dose groups after 24 h treatment was measured by immunofluorescence.
The number of apoptotic cells of 3.13 x 10(-6) mol/L group was more than the control group examined by fluorescence microscope, and the number of dead cells of 5.00 x 10(-5) mol/L group was more than the control group. The apoptotic rates of PC12 cells was higher in the 3.13 x 10(-6) mol/L group than the control group measured by FCM, and the dead rates of PC12 cells was higher in the 5.00 x 10(-5) mol/L group than the control group (P < 0.05). Immunofluorescence cytochemistry experiment demonstrated that the level of Bcl-2 expression was significantly lower in the 3.13 x 10(-6) mol/L group than the control group.
At low dosage, fipronil increases the apoptotic rates of PC12 cells possibly by decreasing the expression of Bcl-2 protein while at high dosage, fipronil only increases the amount of necrotic cells.
研究氟虫腈对PC12细胞的凋亡作用及其机制。
采用形态学方法检测氟虫腈3.13×10(-6)、1.25×10(-5)和5.00×10(-5)mol/L三个剂量组作用24 h后对PC12细胞凋亡和坏死的影响，并用流式细胞仪（FCM）检测凋亡率。采用免疫荧光法检测氟虫腈3.13×10(-6)、1.25×10(-5)和5.00×10(-5)mol/L三个剂量组作用24 h后PC12细胞中Bcl-2蛋白的表达。
荧光显微镜下观察，3.13×10(-6)mol/L组凋亡细胞数多于对照组，5.00×10(-5)mol/L组死亡细胞数多于对照组。FCM检测结果显示，3.13×10(-6)mol/L组PC12细胞凋亡率高于对照组，5.00×10(-5)mol/L组PC12细胞死亡率高于对照组（P<0.05）。免疫荧光细胞化学实验表明，3.13×10(-6)mol/L组Bcl-2表达水平明显低于对照组。
低剂量时，氟虫腈可能通过降低Bcl-2蛋白表达增加PC12细胞凋亡率；高剂量时，氟虫腈仅增加坏死细胞数量。
