Abstract

The current study has focused on the effects of enniatin B (ENN B, a major mycotoxin produced by Fusarium fungi) on early embryonic development. In in vitro analysis, mouse blastocysts were incubated in medium with ENN B (0–40 μM) or 0.5% DMSO (control group) for 24 hours. In an animal study, blastocysts were collected from mice which were intravenously injected with ENN B (1, 3, 5, and 7mg/kg body weight/day) for 4 days in order to analyze apoptosis and necrosis via Annexin V/PI staining assay; and proliferation using dual differential staining. Exposure to low ENN B concentration (10 μM in vitro and 3 mg/kg/day in vivo) promoted Reactive Oxygen Species (ROS) generation and apoptosis in the Inner Cell Mass (ICM), the mass of cells inside the blastocyst, impairing post-implantation development alone. On the other hand, exposure to a higher ENN B concentration (40 μM in vitro and 7 mg/kg/day in vivo) induced ROS generation and decreased in intracellular ATP which encouraged necrotic processes in both trophectoderm (TE) and ICM of blastocysts leading to impaired implantation and post-implantation development. Moreover, 5 and 7 mg/kg/day ENN B intraperitoneal injection to female mice for 4 days has caused downregulation of CXCL1, IL-1β and IL-8 expressions and increased ROS generation in the liver of newborn mice. Over all, ENN B can induce apoptosis and/or necrosis depending on the treatment dosage in mouse blastocysts. ENN B-induced necrosis in blastocysts may exert long-term harmful effects on next-generation newborns.

摘要

目前的研究集中在enniatin B（ENN B，一种由镰刀菌产生的主要霉菌毒素）对早期胚胎发育的影响。在体外分析中，小鼠胚泡在含有 ENN B（0-40 μM）或 0.5% DMSO（对照组）的培养基中培养 24 小时。在一项动物研究中，从小鼠中收集囊胚，这些小鼠静脉注射 ENN B（1、3、5 和 7mg/kg 体重/天）4 天，以通过膜联蛋白 V/PI 染色分析分析细胞凋亡和坏死; 和增殖使用双重差异染色。暴露于低 ENN B 浓度（体外10 μM 和体内3 mg/kg/天）) 促进内细胞团 (ICM) 中的活性氧 (ROS) 生成和细胞凋亡，内细胞团是囊胚内的细胞团，仅损害植入后发育。另一方面，暴露于较高的 ENN B 浓度（体外40 μM 和体内7 mg/kg/天）会诱导 ROS 产生并减少细胞内 ATP，这会促进滋养外胚层 (TE) 和囊胚 ICM 的坏死过程，导致损害植入和植入后的发育。此外，向雌性小鼠腹腔注射 5 和 7 mg/kg/天 ENN B 4 天已引起CXCL1、IL-1β和IL-8的下调新生小鼠肝脏中 ROS 的表达和增加的产生。总之，ENN B 可以根据小鼠胚泡的治疗剂量诱导细胞凋亡和/或坏死。ENN B 诱导的囊胚坏死可能对下一代新生儿产生长期的有害影响。