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% under 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 various 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Leydigcell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20μmol/L浓度的ZEN体外作用于猪Leydigcell,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义(P<0.01),且存在明显的剂量-效应关系;各剂量组对应的拖尾细胞尾长(Taillength)分别为57.60±4.78、57.75±6.25、78.97±5.83、100.50±6.94和146.83±12.31μm,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,5μmol/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。
基金
江苏省普通高校自然科学基金(08KJD230002)资助
扬州大学科技创新培育基金资助~~