Guo Hong-Wei, Chang Juan, Wang Ping, Yin Qing-Qiang, Liu Chao-Qi, Xu Xiao-Xiang, Dang Xiao-Wei, Hu Xiao-Fei, Wang Quan-Liang
College of Animal Science and Technology, Henan Agricultural University, Zhengzhou, 450046, China.
Henan Delin Biological Product Co., Ltd, Xinxiang, 453000, China.
AMB Express. 2021 Mar 1;11(1):35. doi: 10.1186/s13568-021-01196-7.
Aflatoxin B (AFB) is one of the most dangerous mycotoxins for humans and animals. This study aimed to investigate the effects of compound probiotics (CP), CP supernatant (CPS), AFB-degradation enzyme (ADE) on chicken embryo primary intestinal epithelium, liver and kidney cell viabilities, and to determine the functions of CP + ADE (CPADE) or CPS + ADE (CPSADE) for alleviating cytotoxicity induced by AFB. The results showed that AFB decreased cell viabilities in dose-dependent and time-dependent manners. The optimal AFB concentrations and reactive time for establishing cell damage models were 200 µg/L AFB and 12 h for intestinal epithelium cells, 40 µg/L and 12 h for liver and kidney cells. Cell viabilities reached 231.58% (p < 0.05) for intestinal epithelium cells with CP addition, 105.29% and 115.84% (p < 0.05) for kidney and liver cells with CPS additions. The further results showed that intestinal epithelium, liver and kidney cell viabilities were significantly decreased to 87.12%, 88.7% and 84.19% (p < 0.05) when the cells were exposed to AFB; however, they were increased to 93.49% by CPADE addition, 102.33% and 94.71% by CPSADE additions (p < 0.05). The relative mRNA abundances of IL-6, IL-8, TNF-α, iNOS, NF-κB, NOD1 (except liver cell) and TLR2 in three kinds of primary cells were significantly down-regulated by CPADE or CPSADE addition, compared with single AFB group (p < 0.05), indicating that CPADE or CPSADE addition could alleviate cell cytotoxicity and inflammation induced by AFB exposure through suppressing the activations of NF-κB, iNOS, NOD1 and TLR2 pathways.
黄曲霉毒素B(AFB)是对人类和动物最危险的霉菌毒素之一。本研究旨在探讨复合益生菌(CP)、CP上清液(CPS)、AFB降解酶(ADE)对鸡胚原代肠上皮细胞、肝和肾细胞活力的影响,并确定CP+ADE(CPADE)或CPS+ADE(CPSADE)减轻AFB诱导的细胞毒性的作用。结果表明,AFB以剂量和时间依赖性方式降低细胞活力。建立细胞损伤模型的最佳AFB浓度和反应时间为肠上皮细胞200μg/L AFB和12小时,肝和肾细胞40μg/L和12小时。添加CP后肠上皮细胞的细胞活力达到231.58%(p<0.05),添加CPS后肾和肝细胞的细胞活力分别达到105.29%和115.84%(p<0.05)。进一步的结果表明,当细胞暴露于AFB时,肠上皮细胞、肝和肾细胞活力显著降低至87.12%、88.7%和84.19%(p<0.05);然而,添加CPADE后细胞活力提高到93.49%,添加CPSADE后分别提高到102.33%和94.71%(p<0.05)。与单一AFB组相比,添加CPADE或CPSADE后,三种原代细胞中IL-6、IL-8、TNF-α、iNOS、NF-κB、NOD1(肝细胞除外)和TLR2的相对mRNA丰度显著下调(p<0.05),表明添加CPADE或CPSADE可通过抑制NF-κB、iNOS、NOD1和TLR2途径的激活来减轻AFB暴露诱导的细胞毒性和炎症。