呕吐毒素、伏马毒素B1单一及联合暴露引起猪肠道炎症的细胞焦亡机制研究

有研究表明，猪饲料中呕吐毒素（DON）和伏马毒素B1（FB1）的联合污染非常严重。DON和FB1进入体内后首先作用于消化道导致炎症，但关于这两种毒素单一及联合暴露引起肠道炎症的作用及其机制还不清楚。本课题拟进行动物试验，通过荧光定量PCR、免疫印迹、病理组织切片、ELISA等方法，研究DON、FB1单一及联合暴露对猪肠道炎症产生的影响及其可能的细胞焦亡机制；然后以猪小肠上皮细胞IPEC-J2为模型，研究DON、FB1单一及联合暴露对细胞炎性因子前体和炎性因子的影响，通过CRISPR/Cas9敲除、RNA干扰、荧光定量PCR、免疫印迹和电镜等方法研究NLRP3、Caspase-1、GSDMD等相关焦亡蛋白在其中的作用，以阐明DON、FB1单一及联合暴露引起猪肠道炎症的细胞焦亡机制。本课题的开展可揭示DON、FB1单一及联合暴露引起肠道炎症的致病机理，并为饲料中毒素限量标准的制定提供科学参考。

It has been reported that the combined contamination of vomiting toxin DON and fumonisin FB1 is very serious in pig feeds currently. It is known that DON and FB1 firstly acts on the digestive tract after entering into the body, and cause intestinal inflammation, but the effects of the two toxins alone or in combination on intestinal inflammation and its mechanisms remain unclear. This study is intended to conduct piglet experiments, using real-time PCR, immunoblotting, pathological tissue section, ELISA and other methods to evaluate the effects of DON, FB1 alone or combined exposure on porcine intestinal inflammation and its possible mechanism related to pyroptosis. Porcine intestinal epithelial cell IPEC-J2 was also used as a model in vitro to study the effects of DON, FB1 alone or combined exposure on inflammatory factors and pro-inflammatory factors. The role of pyroptosis in porcine intestinal inflammation caused by DON and FB1 alone or combined exposure and its mechanisms will be further clarified by determining the production and activation of NLRP3, caspase 1, GSDMD and other pyroptosis-related proteins through CRISPR/Cas9 knockout, RNA interference, real-time PCR, immunoblotting and electron microscopy, respectively. This study can reveal the pathogenic mechanism of intestinal inflammation caused by single and combined exposure of DON and FB1, and provide data reference for toxin limit standards in pig feed.