丁氟螨酯对SH-SY5Y细胞的毒性作用及其机制

发布时间：2018-06-01 05:33

  本文选题：丁氟螨酯 + SH-SYY细胞　； 参考：《中国药理学与毒理学杂志》2017年04期

【摘要】：目的研究丁氟螨酯对人经神经母细胞瘤细胞SH-SY5Y细胞的毒性作用及其机制。方法加入丁氟螨酯0.03,0.06,0.125,0.25,0.5,1,2,2.6,4,6,8和16 mmol·L~(-1)处理SH-SY5Y细胞48 h,MTT法测定细胞存活;DCFH-DA荧光探针标记法检测细胞内活性氧(ROS)水平;JC-1标记法检测细胞线粒体膜电位;Hoechst 33258染色观察细胞核形态;碘化丙啶(PI)染色和流式细胞仪检测细胞周期和细胞凋亡;Western蛋白印迹法检测磷酸化P38蛋白(p-P38)和磷酸化Jun激酶(p-JNK)表达水平。结果与溶剂(DMSO)对照组相比,共孵育48 h后,丁氟螨酯≥0.06 mmol·L~(-1)时可降低细胞存活率(P0.05),且随浓度增高细胞存活率有降低趋势,IC_(50)为2.6 mmol·L~(-1);丁氟螨酯1,2,4和6 mmol·L~(-1)组细胞ROS水平升高(P0.01),线粒体膜电位下降(P0.01)。Hoechst 33258染色结果显示,丁氟螨酯2,4和6 mmol·L~(-1)组SH-SY5Y细胞出现颗粒状荧光,细胞核固缩和崩解;流式细胞仪检测结果显示,丁氟螨酯2,4和6 mmol·L~(-1)组细胞凋亡率由DMSO对照组的(0.7±0.1)%分别上升至(6.7±0.1)%,(72.4±8.6)%和(90.7±3.2)%(P0.01);丁氟螨酯4和6 mmol·L~(-1)组G_1期细胞较DMSO对照组明显增加(P0.01);Western蛋白印迹结果表明,丁氟螨酯4和6 mmol·L~(-1)组p-JNK表达均升高(P0.01),丁氟螨酯6 mmol·L~(-1)组p-P38表达水平增加(P0.01)。结论丁氟螨酯可能通过氧化损伤、激活P38蛋白和JNK蛋白诱导SH-SY5Y细胞G_1期阻滞和凋亡。
[Abstract]:Objective to study the toxic effect of bufluride on human transneuroblastoma (SH-SY5Y) cells and its mechanism. Methods SH-SY5Y cells were treated with 0.03 ~ 0.06 ~ (0.125) ~ (0.25) ~ (0.125) ~ 0.25 ~ 0.25 ~ (5) and 16 mmol 路L ~ (-1) ~ (-1) respectively. The viability of SH-SY5Y cells was detected by fluorescence probe labeling method. The level of reactive oxygen species (Ros) was detected by JC-1 labeling method. The cell mitochondria membrane potential was detected by Hoechst 33258 staining and the nuclear morphology was observed by Hoechst 33258 staining. The expression of phosphorylated P38 protein (p-P38) and phosphorylated Jun kinase (p-JNKK) were detected by Western blot and flow cytometry. Results compared with the control group, the control group was incubated for 48 hours. The cell survival rate decreased with the increase of cell concentration, and the cell survival rate decreased with the increase of concentration. The cell ROS level in the groups of bufluroate 鈮，

本文编号：1963027