玉米赤霉烯酮对猪睾丸间质细胞DNA损伤作用的彗星试验

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20μmol/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(Tail length)分别为57.60±4.78、57.75±6.25、78.97±5.83、100.50±6.94和146.83±12.31μm,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。

Single Cell Gel Electrophoresis Assay of Porcine Leydig Cell DNA Damage Induced by Zearalenone

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 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.