Ren Zhihua, Deng Huidan, Deng Youtian, Liang Zhen, Deng Junliang, Zuo Zhicai, Hu Yanchun, Shen Liuhong, Yu Shumin, Cao Suizhong
College of Veterinary Medicine, Sichuan Agricultural University, Sichuan Province Key Laboratory of Animal Disease & Human Health, Key Laboratory of Environmental Hazard and Human Health of Sichuan Province, Chengdu, 611130, China;.
College of Veterinary Medicine, China Agricultural University; Beijing, 100094, China.
Exp Toxicol Pathol. 2017 Oct 2;69(8):612-617. doi: 10.1016/j.etp.2017.05.008. Epub 2017 Jun 3.
Deoxynivalenol (DON) and zearalenone (ZEA) are the two most common mycotoxins in animal feed. In this study, we examined oxidative injury and apoptosis of porcine splenic lymphocytes induced by DON or ZEA and their combination in vitro. Based on IC values, porcine splenic lymphocytes were treated with 0.06, 0.3, 1.5, and 7.5μg/mL DON, 0.08, 0.4, 2, and 10μg/mL ZEA, or both DON and ZEA at 0.06 and 0.08μg/mL, 0.3 and 0.4μg/mL, and 1.5 and 2μg/mL, respectively. After 48h of DON and/or ZEA exposure, the cells were analyzed for antioxidant functions, apoptosis, and mRNA and protein expression of apoptosis-related genes p53, Bcl-2, Bax, caspase-3, and caspase-8 to determine their apoptosis and oxidative damage effects and mechanisms. The results showed that, compared with the control group, SOD, CAT, GPx, GSH, and Bcl-2 mRNA and protein expression levels were significantly reduced in exposed groups (P<0.05 or P<0.01). Furthermore, MDA contents, apoptosis rates, and p53, Bax, caspase-3 and caspase-8 protein and mRNA expression levels were increased significantly (P<0.01). The effects of DON and ZEA were dose dependent and synergistic in combination. These data suggest that DON and ZEA induce oxidative damage and apoptosis of porcine splenic lymphocytes.
脱氧雪腐镰刀菌烯醇(DON)和玉米赤霉烯酮(ZEA)是动物饲料中两种最常见的霉菌毒素。在本研究中,我们检测了DON或ZEA及其组合在体外诱导猪脾淋巴细胞的氧化损伤和凋亡情况。根据半数抑制浓度(IC)值,用0.06、0.3、1.5和7.5μg/mL的DON、0.08、0.4、2和10μg/mL的ZEA,或分别用0.06和0.08μg/mL、0.3和0.4μg/mL以及1.5和2μg/mL的DON和ZEA组合处理猪脾淋巴细胞。在DON和/或ZEA暴露48小时后,分析细胞的抗氧化功能、凋亡情况以及凋亡相关基因p53、Bcl-2、Bax、caspase-3和caspase-8的mRNA和蛋白表达,以确定它们的凋亡和氧化损伤作用及机制。结果表明,与对照组相比,暴露组的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)、谷胱甘肽(GSH)以及Bcl-2的mRNA和蛋白表达水平显著降低(P<0.05或P<0.01)。此外,丙二醛(MDA)含量、凋亡率以及p53、Bax、caspase-3和caspase-8的蛋白和mRNA表达水平显著升高(P<0.01)。DON和ZEA的作用呈剂量依赖性且联合时具有协同作用。这些数据表明,DON和ZEA可诱导猪脾淋巴细胞的氧化损伤和凋亡。