桑园常用杀菌剂多菌灵诱导鼠源神经细胞PC12凋亡的研究

A Study on Mouse Neural Cell PC12 Apoptosis Induced with Carbendazim Commonly Used in Mulberry Field

多菌灵是桑园病虫害防治的常用农药,并用于家蚕微粒子病的防治。研究多菌灵对哺乳动物神经系统的毒性损伤作用及可能的作用机制,为避免蚕区桑园用药污染环境的危害提供理论依据。采用MTT法、流式细胞计数法测定用质量浓度为50、100和200μg/m L的多菌灵药液分别与鼠源神经细胞PC12共同孵育48 h后,可极显著抑制PC12细胞的增殖（P〈0.01）,诱导PC12细胞凋亡,特别是早期凋亡（P〈0.01）。实时荧光定量PCR检测经50μg/m L多菌灵药液处理后培养48 h的PC12细胞内,促凋亡蛋白基因Bax和凋亡相关基因Cyt-C、Fas、Caspase-8、Caspase-9的表达水平极显著上调（P〈0.01）;同时检测50μg/m L多菌灵药液处理组PC12细胞中Caspase-3的酶活性也极显著提高（P〈0.01）。研究结果显示50~200μg/m L多菌灵药液对哺乳动物神经细胞具有一定的毒性作用,推测其可能通过启动死亡受体介导途径和线粒体介导途径诱导神经细胞凋亡,因此在桑园施用多菌灵时,应注意防护以及远离居住区。

Carbendazim is a common pesticide used in mulberry field and was used to control pebrine disease. To provide theoretical basis for avoiding environmental pollution in mulberry fields,this paper studied the toxic injury of carbendazim on nervous system of mammals and its underlying mechanism（ s）. MTT and flow cytometry analysis results showed that addition of carbendazim into PC12 cells with mass concentrations of 50,100 and 200 μg / mL respectively could extremely significantly inhibit the proliferation of PC12 cells（ P 0. 01）,induce the apoptosis especially the early apoptosis of PC12 cells（ P 0. 01）. Real-time fluorescent quantitative（ qRT-PCR） results showed that the mRNA transcriptional levels of pro-apoptotic protein gene Bax and apoptosis-associated genes Cyt-C, Fas, Caspase-8 and Caspase-9 were extremely significantly increased in PC12 cells at 48 h after treatment with 50 μg / mL carbendazim（ P 0. 01）. In addition,the activity of Caspase-3 in 50 μg / mL carbendazim-treated PC12 cells was also extremely significantly increased（ P 0. 01）. These results indicated that carbendazim of 50 to 200 μg / mL has certain cytotoxic effect on mammal neural cells. It could induce apoptosis through activate the death receptor-mediated and mitochondria-mediated signal transduction pathways. Its application should be far away from residential areas and proper protection should be taken when used in mulberry fields.