玉米赤霉烯酮对猪睾丸间质细胞DNA损伤作用的彗星试验

摘要

[Objective] This study aimed to investigate the effect of zearalenone (ZEN) on DNA damage of porcine leydig cells. [Method] Porcine leydig cells cultured in vitro were collected to determine the median lethal dose (LD50 ) of ZEN with tetrazolium-based colorimetric assay (MTT assay). Comet assay was carried out to detect the DNA damage of porcine leydig cells exposed to at 0 (negative group) ,1,5, 10, 20, 40 μmol/L of ZEN. [Result] The percentage of cell tail was 16.67% , 34.00% , 40.67% , 52.00% and 64.67% after exposed to 0, 1 , 5, 10 and 20 μmol/L of ZEN, respectively; the differences between the percentages of cell tail in various experimental groups had extremely significant statistical significance compared with the negative group (P<0. 01) , showing a significant dose-effect relationship; Tail length in various groups was 57.60 ±4.78, 57.75 ±6.25, 78.97 ±5.83, 100.50 ±6. 94 and 146.83 ±12.31 μm, respectively; Tail DNA % in vari-ous groups was 21.29 ±2.25% , 22.24+2.43%, 31,21 ±6.27% , 37.45 ±4.33% and 60. 68 ±9.83%, respectively; Tail length and Tail DNA % in experimental groups with ZEN concentration above 5 μmol/L showed significant differences (P <0. 05) compared with the negative group, which showed an upward trend with the increase of ZEN concentration. [Conclusion ] ZEN has genotoxic effect on porcine leydig cells, which can cause DNA damage, with a significant dose-effect relationship.%[目的]观察玉米赤霉烯酮(ZEN)对猪睾丸间质细胞( Leydig cell)的DNA损伤效应.[方法]以体外培养的猪Leydig cell为材料,用四氮唑蓝比色分析法(MTT法)测定ZEN对离体培养的猪睾丸间质细胞的半数致死浓度,选用0(对照组)、1、5、10和20 μmol/L浓度的ZEN体外作用于猪Leydig cell,通过彗星试验观察了ZEN对猪Leydig cell DNA的损伤效应.[结果]ZEN浓度为0、1、5、10和20μmol/L时,所造成的细胞拖尾率分别为16.67％ 、34.00％、40.67％、52.00％和64.67％,各染毒组细胞拖尾率与对照组比较,差异均有极显著统计学意史(P＜0.01),且存在明显的剂量-效应关系；各剂量组对应的施尾细胞尾长(Tail length)分别为57 60±4.78、57.75±6.25、78.97±5.83、100.50±6.94和146.83±12.31 μm,尾部DNA含量(Tail DNA％)分别为21.29±2.25％、22.24±2.43％、31.21±6.27％、37.45±4.33％和60.68±9.83％,与对照组比较,5μ mol/L及以上ZEN浓度染毒组的Tail length和Tail DNA％均有显著性差异(P＜0.05),且随ZEN浓度增加呈上升趋势.[结论]ZEN对猪Leydig cell存在遗传毒性,可以损伤Leydig cell的DNA,且有明显的剂量-效应关系.