辣蓼黄酮对病毒感染免疫细胞Nrf2氧化应激信号通路相关LncRNA及组蛋白乙酰化修饰调控的分子机制

本项目拟选择小鼠、猪的免疫细胞为研究对象，以免疫药理学和自由基生物学理论结合分子生物学技术，探讨辣蓼黄酮对病毒感染免疫细胞Nrf2氧化应激信号通路相关LncRNA和组蛋白乙酰化修饰调控的分子机制。分离提取及纯化辣蓼黄酮；选择猪伪狂犬病毒，通过体内外实验观察病毒感染对免疫细胞内活性氧水平影响及辣蓼黄酮的调节作用；从Nrf2氧化应激信号通路相关LncRNA差异表达方面，观察LncRNA在病毒感染免疫细胞氧化应激中的作用及其与组蛋白乙酰化修饰相关性；从LncRNA表达、组蛋白及乙酰化相关酶和活性氧等的相关基因和蛋白表达方面，观察辣蓼黄酮对病毒感染免疫细胞Nrf2氧化应激信号通路相关LncRNA和组蛋白乙酰化修饰的调控机制；从LncRNA-Y定位调控的靶基因表达方面，探讨辣蓼黄酮对Nrf2氧化应激信号通路相关LncRNA调控机制，可为发现黄酮类药物作用靶点提供依据，具有重要学术意义和潜在应用价值。

The objective of this project is to investigate molecular regulatory mechanisms of active flavonoids isolated from Polygonum hydropiper L. on LncRNA related with Nrf2 oxidative stress signal pathway and histone acetylation modification in PRV infected immunocytes of mice and pigs in vivo and in vitro. The study involves an intersection field of new technology of molecular biology and basic theories of immunopharmacology and free radical biology. In this project, regulatory effect of flavonoids of Polygonum hydropiper L on levels of reactive oxygen species will be observed in vivo and in vitro in immunocytes infected by PRV. The actions of LncRNA on the oxidative stress related with Nrf2 signal pathway and relationship with histone acetylation modification will be investigated by measuring differential expression of LncRNA in immunocytes infected by PRV. The regulatory mechanism of the flavonoid on LncRNA related with Nrf2 signal pathway and histone acetylation modification in PRV infected immunocytes in vivo and in vitro will be observed by investigating gene expression and protein expression of LncRNA, histones and histone related enzymes as well as of reactive oxygen species. The regulatory mechanism of the flavonoid on LncRNA related with Nrf2 oxidative stress signal pathway will be elucidated by observing expression of target genes of LncRNA-Y in the immunocytes infected by the virus. The achievements will be of benefit to find a new drug target of flavonoids and control virus infections in animals.