氰戊菊酯对Tn细胞系P450及蛋白质谱表达的影响

Effects of Fenvalerate on Cytochrome P450 and the Protein Map of Tn Cells

研究氰戊菊酯对粉蚊夜蛾(Trichoplusia ni,Tn)细胞系细胞色素P450(P450s)的诱导作用及蛋白质谱表达的影响．用MTT法和台盼兰法观察不同浓度氰戊菊酯处理Tn细胞12 h后细胞的存活率；光谱测定法测定P450s含量；以12．5μmol/L氰戊菊酯处理Tn细胞12 h,通过二维液相色谱分离技术分析处理前后Tn细胞系蛋白质谱的变化．结果显示：当氰戊菊酯处理浓度高于17．5μmol/L时,Tn细胞形态发生明显变化,且细胞死亡率显著高于对照组(P＜0．01)．P450s的含量有随氰戊菊酯浓度增加而升高的趋势,并在氰戊菊酯浓度为12．5μmol/L时达到最高(8．562nmol/mg protein)；P450s的含量亦有随处理时间延长而升高的趋势,且在12 h时达到最高(5．28±1．14 nmol/mg protein)．通过二维液相色谱分离技术得到124个差异组分,其中42个组分上调,82个组分下调．结果表明：氰戊菊酯对P450s确有诱导作用,且氰戊菊酯诱导前后Tn细胞中蛋白的变化可能与P450s诱导途径中某些受体的变化有关．

To learn the effects of Fenvalerate on P450s and protein expression in Tn （Trichoplusia hi） cells, the cells were treated with different concentrations of fenvalerate for 12 h to observe the INability of cells by Trypan Blue and MTT assay. P450s Contends were determined by spectroscopic analysis. Protein maps of controlled and treated groups were analyzed by two-dimensional liquid phase chromatographic fractionation （PF-2D） after treated Tn cells with 12.5 μM fenvalerate for 12 h. Significant differences in characteristic and mortality of cells were found while the concentration of fenvalerate was over 17.5 μM. P450s activity trended to be enhanced with the increase of time or dose of fenvalerate. The activity of P450s reached a maximum at the dose of 12.5μM and the time of 12 h, and the value of maximum is （8. 562 ± 3.46） nmol/ mg and （5.28±1. 14） nmol/mg protein. 124 differences points were detected between the 2D maps of treated and controlled groups. 42 points were up-regulated and 82 were down-regulated. Fenvalerate can induce P450s in Tn cells and those proteins changed after treated With fenvalerate might be related to some receptors in the P450s inducing pathway.