有机磷农药辛硫磷对家蚕卵巢培养细胞DNA损伤的单细胞凝胶电泳检测

Single Cell Gel Electrophoretic Assay on DNA Dam age of Silkworm BmN Cells Caused by Organophosphorus Pesticide Phoxim

采用单细胞凝胶电泳技术检测农药对家蚕卵巢培养细胞(Bm N)DNA的损伤作用,从细胞毒理学方面确证农药对家蚕的残留毒性,并探讨家蚕培养细胞作为农药危害及残留检测生物标志物的可行性。以有机磷杀虫剂辛硫磷以及Bm N细胞供试,用0.05 mg/L辛硫磷药液(约为家蚕5龄第1天幼虫24 h LC50的1/100)处理正常Bm N细胞,24 h后观察到处理组Bm N细胞出现明显的凋亡现象,通过单细胞凝胶电泳检测到Bm N细胞的DNA条带有95.43%出现拖尾现象。试验结果表明Bm N细胞对辛硫磷非常敏感,低浓度辛硫磷即会对细胞DNA造成严重损伤,该项技术可用于农药对家蚕的危害及残留毒性鉴定,同时也说明Bm N细胞可以作为农药的细胞毒性研究材料。

In present study, the damage of pesticide to silkworm （Bombyx mori） ovarian culture cell （BmN） DNA was detected by single cell gel electrophoresis to confirm residual toxicity of pesticide to silkworm from the aspect of cytotoxi- cology. Besides, we discussed the feasibility of using silkworm culture cell as biomarker for the detection of pesticide damage and its residue. The organophosphorus pesticide phoxim and BmN were used as materials. After being treated with 0.05 mg/L phoxim （the concentration was about 1/100 of the 24 h LCso of phoxim to day 1 silkworm larvae of the 5th instar） for 24 h, cell apoptosis was observed obviously in the treatment group. Single cell gel electrophoresis showed that 95.43% of bands of BmN cell DNA exhibited tailing phenomenon. The results indicated that BmN cells are very sen- sitive to phoxim, and low concentration phoxim will cause serious damage to cell DNA. The established technology can be used for identification of damage and residual toxicity of pesticide to silkworm, and BmN cells can be used as material for studying pesticide cytotoxicity.