适配体靶向干扰体内玉米赤霉烯酮作用机制

Zearalenone in feed and feedstuff is ubiquitous and difficult to remove. Once enrichment of animals, it causes serious damage to the body. Methods of removal zearalenone in vivo need to be improved. Aptamers are DNA or RNA oligonucleotides capable of binding different classes of targets with high affinity and selectivity. It has good prospects for development as a feed mycotoxin adsorbent. In this proposal, apatmers, combined with zearalenone, will be screened using SELEX method aptamer in vitro. By cell culture, aptamers will be screened, which are inhibiting bind of zearalenone and estrogen receptor. Molecular simulation will help to confirm interference mechanism of aptamer. After modified by polyethylene glycol, apatmer will be investigated the adsorption effects of zearalenone by detecting zearalenone residues using High Performance Liquid Chromatography, determinating gene expression and protein expression of estrogen receptor levels. Our results will apply the mechanisms of removal mycotoxin in vivo, and provide the basis for the development of efficient and stable new adsorbents.

饲料及原料中玉米赤霉烯酮污染普遍存在且难以清除，富集至动物体内，对机体产生严重危害。目前体内去除玉米赤霉烯酮方法的高效性和稳定性还有待提高。核酸适配体（aptamer）是一类能识别特定靶分子的单链DNA或RNA，其高亲和力、强专一性的特点，可作为饲料霉菌毒素吸附剂进行开发。本项目采用SELEX方法体外筛选能与玉米赤霉烯酮结合的适配体；通过细胞培养筛选出能够抑制玉米赤霉烯酮与雌激素受体结合的适配体；进行生物信息学的分子模拟，确定适配体的干扰机制；经聚乙二醇修饰后稳定性增加的适配体进行应用，通过高效液相色谱测定玉米赤霉烯酮残留量、基因表达和蛋白表达测定雌激素受体水平，考察其在体内吸附玉米赤霉烯酮的效果。研究结果将不仅为霉菌毒素体内去除机制拓展思路，同时为高效、稳定的新型吸附剂的开发奠定基础。

饲料及原料中玉米赤霉烯酮污染普遍存在且难以清除，富集至动物体内，对机体产生严重危害。目前体内去除玉米赤霉烯酮方法的高效性和稳定性还有待提高。核酸适配体（aptamer）是一类能识别特定靶分子的单链DNA或RNA，其高亲和力、强专一性的特点，可作为饲料霉菌毒素吸附剂进行开发。本项目采用非固定靶标法的SELEX筛选技术，得到了与玉米赤霉烯酮高亲和力（15.2±3.4 nmol/L）和特异性的适配体，并进行了生物信息学分析，确定了适配体与玉米赤霉烯酮的关键结合位点；制备纳米金溶液，结合此适配体，初步开发出饲料中玉米赤霉烯酮含量的检测方法，检测的线性范围为4~128 ng/mL，检出限为4 ng/mL，并能够有效检测污染饲料中玉米赤霉烯酮的含量；玉米赤霉烯酮处理小鼠原代子宫细胞和小鼠的实验表明，适配体能够对体外细胞和体内子宫组织起到抗氧化和抗凋亡的保护作用。研究结果为玉米赤霉烯酮的防控监测提供了支撑，为霉菌毒素体内去除机制的探讨拓展了思路，为高效、稳定的新型吸附剂的开发奠定了基础。