硒蛋白S在硒减轻猪赭曲霉毒素A中毒中的作用与机制研究

赭曲霉毒素A(OTA）是广泛存在于饲料中的一种具有很强毒性的霉菌毒素，近年研究表明硒能保护细胞免遭OTA诱发的氧化损伤，改善受损的细胞免疫功能，减轻对猪的毒害作用，但机制不清。硒蛋白是硒在体内发挥作用的主要载体，由于其活性中心硒代半胱氨酸由UGA编码，因而体外表达困难，影响对其进行深入研究，尤其在猪上研究极少。新近发现的人硒蛋白S（SelS）已被证实参与氧化还原平衡的调控。本项目主要是应用RACE、Western-blot、RNAi、流式细胞术和激光共聚焦等技术，对猪SelS进行全基因序列克隆、鉴定与生物信息学分析，构建SelS基因低表达与过表达猪PK15肾细胞和原代T淋巴细胞株，从体外到体内研究猪SelS抗氧化功能、免疫调节功能、在硒减轻OTA诱发的细胞毒性中的作用和在硒改善OTA诱发的免疫抑制中的作用以及MAPK信号通路机制。本项目对揭示猪SelS功能与防控猪赭曲霉毒素中毒有重要意义。

Ochratoxin A (OTA) is a highly toxic mycotoxin and widely present in feeds. Recent studies have shown that selenium can protect cells from oxidative damage, improve damaged cellular immune function and reduce the toxic effects induced by OTA in pigs, but the mechanism is unclear. Selenoprotein is a main carrier by which selenium exerts the biological role in the body. Becase selenocysteine, the active site of selenoprotein, is encoded by the UGA, expression in vitro is very difficult and its in-depth research is influenced, particularly in pigs. Selenoproteins s (SelS), a newly discovered selenoprotein in humans, has been confirmed to participate in the regulation of redox balance. The main work of this project is as follows: whole genome sequence cloning, identification and bioinformatics analysis of SelS in pigs; construction of pig PK15 kidney cell lines and primary T lymphocyte cell lines with low expression and overexpression of SelS gene; in vitro and in vivo studies on swine SelS antioxidant function, SelS immune function, the role of SelS on selenium alleviation of the OTA -induced cytotoxicity, the role of SelS on selenium improvement of OTA -induce immunosuppression and the mechanisms of MAPK signaling pathway by RACE, Western-Blot, RNAi, flow cytometry and laser confocal microscopy etc. technology. The project will be helpful to reveal the function of SelS and control of ochratoxin poisoning in pigs.