白藜芦醇对伪狂犬病毒感染妊娠母猪的作用效果及其机制研究

Pseudorabies virus (Pseudorabies, virus, PRV) can infect a variety of animals. As its natural host, high morbidity and mortality often occurs after infection in pigs. PRV infection can result in severe reproductive failure in pregnant sows, characterized by abortion, still birth, mummification and infertility. Studies indicate that PRV virus can invade the placenta and break the placental barrier, causing fatal infection with clinical consequences of PRV infection. At present, it is lack of effective preventive and cure ways to this pathogen, especially its variant strain, and the PRV infection is still prevalent in our country. Results from in vivo and in vitro studies have shown that plant extract resveratrol has a very good antiviral activity. Our previous studies have also shown that resveratrol exhibits obvious antiviral effect in mice with PRV infection induced reproductive failure. To further explore its effect on PRV infection in pregnant sows and its possible mechanism, firstly, an animal model with PRV infection will be developed, and the effect of resveratrol will be investigated in vivo. To mimic the local infection of PRV in endometrium or placenta tissues, a three-dimensional tissue culture system will be established, which is used to explore the antiviral effects of resveratrol in local. Moreover, three types of porcine primary cells including endometrial cells, placenta trophoblastic cells and umbilical vein endothelial cells will be isolated and cultured. Resveratrol will be administrated in PRV infected cells, cell functions will be detected to check its effects in vitro. The results will provide a reference for the prevention and treatment of diseases caused by viral diseases in livestock reproduction, and also provide basic data for the development of resveratrol or related plant extracts as antiviral agents.

伪狂犬病毒（PRV）可引起多种动物感染，猪为其自然宿主，感染后常发生高发病率和死亡率，妊娠母猪感染会导致严重的繁殖障碍，引起流产、死胎、胎儿干尸化及不育等。PRV感染妊娠母猪后，可侵袭胎盘并突破胎盘屏障使胎儿感染而导致临床后果。目前对于该病原特别是其变异株感染的流行还缺乏较理想的防治效果。体内外研究证实白藜芦醇具有很好的抗病毒作用，前期研究表明白藜芦醇对PRV致小鼠繁殖障碍的作用效果明显。本项目旨在进一步研究白藜芦醇对PRV感染妊娠母猪的作用效果，首先将通过动物模型研究白藜芦醇体内对PRV感染的干预作用；并利用三维组织培养模拟PRV局部感染子宫内膜和胎盘组织，从组织层面研究RES对PRV的作用；最后结合原代细胞模型从体外细胞水平研究其作用及其可能作用机制。其结果将为研究病毒性疾病引起的家畜繁殖障碍疾病的防治提供参考，并为开发白藜芦醇或相关植物提取物作为抗病毒中药制剂提供依据。

伪狂犬病毒（PRV）感染是导致猪高发病率和死亡率的重大传染病，妊娠母猪感染会导致严重的繁殖障碍，引起流产、死胎、胎儿干尸化及不育等。PRV感染妊娠母猪后，可侵袭胎盘并突破胎盘屏障使胎儿感染而导致临床后果。目前对于该病原特别是其变异株感染的流行还缺乏较理想的防治效果。体内外研究证实白藜芦醇具有抗病毒作用，且课题组前期研究表明白藜芦醇对PRV导致的小鼠繁殖障碍具有明显的效果。本项目首先通过建立PRV感染的体外细胞模型，并以此为基础探究了PRV感染对宿主细胞增殖和热休克蛋白（HSP）等感染和应激相关蛋白表达的影响，以及白藜芦醇对相关蛋白的调控作用。其次通过母猪饲养试验探究了白藜芦醇对母猪繁殖性能的影响。最后结合母猪饲养试验和小鼠模型，利用组学技术探究了白藜芦醇改善母猪繁殖性能的潜在作用机制。研究结果表明：（1）HSP27、70和90的快速应答可保护细胞免受PRV病毒感染引起的损伤，而白藜芦醇可诱导HSP70上游调控因子SIRT1表达缓解细胞损伤；（2）母猪饲养试验表明饲粮中添加白藜芦醇可以改善母猪胰岛素抵抗，增强抗氧化能力，减少炎症，进而改善母猪的繁殖性能；（3）白藜芦醇可下调Christensenellaceae R-7 group和Ruminococcaceae UCG-005在肠道菌群中的占比，促进SLC2A1、SCL2A3等血管密度和营养转运相关基因的表达，抑制白介素IL-1β和IL-6的产生，进而改善母猪胰岛素抵抗和繁殖性能。上述研究结果揭示了白藜芦醇对PRV病毒感染引起的细胞损伤的潜在保护作用机制，以及白藜芦醇调节肠道菌群，改善母猪代谢，缓解氧化应激、炎症反应及胰岛素抵抗，促进胎盘血液循环，改善母猪繁殖性能的作用。相关结果可为研究病毒性疾病引起的母猪繁殖障碍疾病的防治提供参考，同时也为开发白藜芦醇或相关植物提取物作为改善母猪繁殖性能的中药制剂提供依据。