[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.
为了探讨玉米赤霉烯酮(ZEA)与脱氧雪腐镰刀菌烯醇(DON)联合作用对免疫系统毒性影响及其毒理机制,试验以T淋巴细胞株(CTLL-2)为材料,在利用CCK-8试验确定ZEA、DON对CTLL-2细胞联合染毒浓度的基础上,研究了ZEA与DON联合染毒对CTLL-2细胞凋亡及凋亡相关蛋白Bax、Bcl-2和Caspase-3等表达的影响,试验设对照组、ZEA染毒组(5μg/m L ZEA)、DON染毒组(0.5μg/m L DON)、ZEA+DON联合染毒组(5μg/m L ZEA+0.5μg/m L DON),染毒时间为48 h,用流式细胞术检测细胞的凋亡率,Western-blot检测Bax、Bcl-2和Caspase级联反应相关蛋白的表达情况。结果表明:与对照组相比,各染毒组CTLL-2细胞的凋亡率均极显著升高(P<0.01);各染毒组细胞凋亡相关蛋白Bax/Bcl-2比值和Cleaved Cspase-3、Cleaved Caspase-8、Cleaved Caspase-9等蛋白的表达量均较对照组显著升高(P<0.05或P<0.01);联合染毒组呈现协同效应。说明ZEA与DON联合染毒在诱导免疫细胞凋亡方面发挥协同效应,它们共同存在可以发挥更强的免疫抑制效应;ZEA与DON联合染毒诱导CTLL-2细胞发生凋亡的机制与Bax/Bcl-2上调的线粒体通路密切相关。
