热应激下肉鸡固有免疫应答及白藜芦醇调控的分子机制

At present, differential genes reported were not consistent in the different tissues and organs of chicken under heat stress, so the immune modulation mechanism of chicken under high temperature still need to be explored. Our previous study found that resveratrol could reduce the intestinal inflammatory damage of broiler under heat stress, but the mechanism of resveratrol in alleviating chicken heat stress is still unclear. Therefore, heat stress model with chicken will be established in this study, the differential genes, proteins and signal pathway in innate immune responses in the spleen of chicken with heat stress will be explored by fluorescence quantitative PCR, Western blot, gel migration, flow cytometry, et al., and the regulation effect of resveratrol on the innate immune response of chicken under heat stress will be analyzed and deduced. Thereafter, the target genes, proteins and signal pathways of resveratrol regulating on the innate immune responses in chicken under heat stress will be explored, and the regulation mechanism of resveratrol in chicken under heat stress will be deduced in this study from the view of target gene, protein and cell level and signal pathway, the above results will provide a theoretical basis for the self immune regulation mechanism of poultry under the high temperature environment and the biological function study of resveratrol.

目前鸡对热应激反应不同组织器官中报道的差异基因不一致，高温下鸡自我免疫调控机制需不断完善。我们前期研究发现白藜芦醇能降低热应激下肉鸡肠道炎症损伤，但白藜芦醇缓解鸡热应激的作用机制仍不清楚。本研究建立仔鸡热应激模型，运用荧光定量PCR、免疫印迹、流式细胞术等技术，重点探讨热应激下鸡脾脏内涉及免疫反应的差异基因、蛋白与信号通路，阐明白藜芦醇对热应激下鸡固有免疫应答的调控效果，解析白藜芦醇调控热应激下鸡免疫反应的靶基因、蛋白及信号通路。本研究从基因、蛋白、细胞水平与信号通路角度阐释热应激下鸡固有免疫应答及白藜芦醇调控的分子机制，将为家禽高温环境下自我免疫调控机制及白藜芦醇生物学功能研究提供理论依据。

白藜芦醇是具有抗炎症 抗氧化 抗癌等多种重要功能的天然生物活性物质,亦可作为畜禽抗热应激添加剂在生产中发挥重要作用。本研究建立黄羽肉鸡热应激模型，运用酶联免疫吸附、RT-PCR，免疫印迹、iTRAQ等技术，探讨热应激下鸡脾脏内涉及的差异表达蛋白与信号通路，阐明白藜芦醇对热应激下鸡固有免疫应答的调控效果，解析白藜芦醇调控热应激下鸡免疫反应的靶基因、蛋白及信号通路。本研究从宏观、基因、蛋白水平与信号通路角度阐释热应激下鸡固有免疫应答及白藜芦醇调控的分子机制，将为家禽高温环境下自我免疫调控机制及白藜芦醇生物学功能研究提供理论依据。结果表明：添加适宜白藜芦醇显著提高了热应激黄羽肉鸡平均日增重，从d3开始显著抑制直肠温度的上升；通过调控热应激肉鸡血清抗氧化、内分泌激素、代谢酶等维持机体血清内环境趋于正常水平。白藜芦醇能够显著降低热应激下空肠粘膜HSP70/90和NF-κB mRNA的高度表达，显著提高热应激下空肠mucin-2, sIgA, claudin-1 和ZO-1 mRNA的表达，抑制mucin-4 和claudin-2的表达，降低肠道通透性，修复屏 障功能。ITRAQ数据发现，热应激引起鸡脾脏内差异表达蛋白62个，其中上调6个，下调56个，发挥着不同的生物学功能参与代谢调控。白藜芦醇可以显著抑制热应激诱导的固有免疫模式识别受体的活化，调控NF-κB和MAPK信号通路，抑制炎症反应，有效的维持热应激下黄羽肉鸡脾脏免疫功能。以上结果表明：白藜芦醇可以有效调控血清生理生化指标，修复肠道屏障损伤，抑制固有免疫模式识别受体过度活化，并通过NF-κB和MAPK信号通路，抑制炎症反应，改善生长性能。