Qiu Taotao, Shen Xing, Tian Zhen, Huang Riming, Li Xiangmei, Wang Juan, Wang Rong, Sun Yuanming, Jiang Yiguo, Lei Hongtao, Zhang Huidong
Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University , Guangzhou 510642, China.
Public Health Laboratory Sciences and Toxicology, West China School of Public Health, Sichuan University , No. 17 People's South Road, Chengdu 610041, China.
J Agric Food Chem. 2018 Feb 14;66(6):1543-1550. doi: 10.1021/acs.jafc.7b05385. Epub 2018 Jan 31.
Aflatoxin B (AFB) causes hepatotoxic, genotoxic, and immunotoxic effects in a variety of species. Although various neutralizing agents of AFB toxicity have been studied, the egg yolk immunoglobulin (IgY) detoxification of small molecular toxins and the mechanisms underlying such effects have not yet been reported. In this investigation, anti-AFB IgY against AFB was successfully raised, and a competitive indirect enzyme-linked immunosorbent assay was established with a sensitive half-maximal inhibitory concentration (IC 2.4 ng/mL) and dynamic working range (0.13-43.0 ng/mL). The anti-AFB IgY obtained reduced AFB-induced cytotoxicity, cellular dysfunction, and genotoxicity by protecting cells against apoptotic body formation and DNA strand breaks, preventing G2/M phase cell cycle arrest, reducing AFB-DNA adduct and reactive oxygen species production and maintaining cell migration and invasion and the mitochondrial membrane potential. Anti-AFB IgY significantly inhibited the AFB-induced expression of proteins related to antioxidative, pro-apoptotic, and antiapoptotic processes in a strong dose-dependent manner. These experiments demonstrated that the anti-AFB IgY-bound AFB could not enter cells. This is the first time that IgY has been found to reduce the effects of small molecular toxins, which will be beneficial for the development of antibodies as detoxication agents.
黄曲霉毒素B（AFB）在多种物种中会引起肝毒性、遗传毒性和免疫毒性作用。尽管已经研究了多种AFB毒性的中和剂，但蛋黄免疫球蛋白（IgY）对小分子毒素的解毒作用及其潜在机制尚未见报道。在本研究中，成功制备了抗AFB的IgY，并建立了一种竞争性间接酶联免疫吸附测定法，其具有灵敏的半数抑制浓度（IC 2.4 ng/mL）和动态工作范围（0.13 - 43.0 ng/mL）。所获得的抗AFB IgY通过保护细胞免受凋亡小体形成和DNA链断裂的影响、防止G2/M期细胞周期阻滞、减少AFB - DNA加合物和活性氧的产生以及维持细胞迁移、侵袭和线粒体膜电位，降低了AFB诱导的细胞毒性、细胞功能障碍和遗传毒性。抗AFB IgY以强烈的剂量依赖性方式显著抑制了AFB诱导的与抗氧化、促凋亡和抗凋亡过程相关的蛋白质表达。这些实验表明，与抗AFB IgY结合的AFB无法进入细胞。这是首次发现IgY可减轻小分子毒素的影响，这将有利于开发作为解毒剂的抗体。
