精氨酸与谷氨酸对呕吐毒素所致猪肠上皮细胞损伤的保护机制研究

呕吐毒素广泛污染饲料，是制约猪生长的主要问题，能引起猪肠道上皮细胞形态结构与功能的变化，造成细胞凋亡。精氨酸与谷氨酸是应激条件下猪必需的功能性氨基酸，可促进猪肠上皮细胞蛋白质合成而调控细胞存活。我们前期的动物试验表明，精氨酸与谷氨酸能减轻呕吐毒素对猪肠道的损伤且促进猪的生长。但目前精氨酸与谷氨酸对呕吐毒素所致猪肠道损伤的保护机制尚不清楚。据此，本项目拟以猪肠上皮细胞为模型，分析精氨酸与谷氨酸对呕吐毒素所致猪肠上皮细胞损伤前后细胞的形态结构、增殖与凋亡、细胞内外蛋白质合成、紧密连接蛋白与氨基酸转运载体表达、NF-κB、p38 MAPKs与mTOR通路调控其下游细胞因子表达的影响，并通过磷酸化蛋白质组学、生物信息学分析与试验验证来深入揭示精氨酸与谷氨酸对呕吐毒素所致猪肠上皮细胞损伤的保护机制，为探索猪肠道霉菌毒素损伤的营养调控技术与饲料配方调整技术提供理论依据，对提高猪的生产性能具有重要意义。

Vomitoxin(VT) contamination, which is widespread among swine feed ingredients is known to cause several adverse effects in pigs, including negative structural and functional changes in the intestinal porcine epithelial cells, resulting in cell apoptosis. Arginine and glutamic acid are two functional amino acids whose requirements are increased for pigs under stressful conditions, and are known to promote protein synthesis in intestinal epithelial cells and to regulate cell survival. Our previous animal studies showed that arginine and glutamic acid can relief VT-induced damage in the intestines and promote pig growth. However, the mechanisms by which arginine and glutamic acid ameliorates VT-induced intestinal injury in pigs still remains unclear. This project will focus on the mechanisms through which arginine and glutamic acid relievesthe injurious effects in intestinal pig epithelial cells following VT exposure. Response criteria to be measured includes, cell morphological changes, cell proliferation and apoptosis, cell protein synthesis, cell tight junction proteins, expression of amino acids transporters, and NF-κB, p38 MAPKs and mTOR signaling pathway or inhibition before or after downstream cytokine expression, using phosphoproteomic methods, bioinformatics analysis and experimental verification to investigate the protective mechanisms for arginine and glutamic acid regulation of VT-induced intestinal porcine epithelial cells damage on the system level. It is anticipated that our results will provide a novel theoretical basis for exploring intestinal damage related to nutrition and possible control technologies, including the adjustment of feed formulation for pigs exposed to mycotoxins contamination, thus improving pig performance.