维甲酸相关孤核受体γ t在慢性HBV感染中的作用及机制研究

Th17 cells has been reported to play an important role in the onset and development of chronic hepatitis B.Retinoic acid-related nuclear orphan receptor γt（RORγt） was required for the differentiation of naive CD4+ T helper cells into Th17 cells , our previous study demonstrated for the first time that RORγt maybe act important role in the pathogenesis of CHB,but the exact mechanism is obscure. This study was aimed to evaluate the gene expression of RORγt and its potential role in IL-17 gene knock-out mice with infection of HBV,the relationship of RORγt and disease progression of CHB,and the regulation of RORγt to the associated inflammation cytokines.The mRNA level of RORγt, TNF-α，IFN-γ，IL-6，IL-10 were determined by quantitative real-time polymerase chain reaction (RT-PCR).The protein expression in liver tissue of TNF-α，IFN-γ，IL-6，IL-10 and RORγt were determined by immunohistechemistry.We reduce the expression of RORγt by antisense oligonucleotide interference,to study the change of inflammation of liver tissue.We inferred that the expression of RORγt in HBV infected mice has positive relationship with the inflammation degree and disease progression, suggestiong that RORγt maybe act important role in CHB.The study supply a new clue in the reseach of the pathogenesis of CHB.

本课题运用免疫印迹、逆转录扩增、流式细胞术等手段，重点研究慢性HBV感染状态下IL-17基因敲除小鼠外周血及肝组织中维甲酸相关孤核受体γt（RORγt）和相关炎性细胞因子TNF-α、IFN-IL-6、IL-10等的表达变化规律，阐明RORγt对慢性HBV感染状态下炎症反应的调控机制；通过IL-17基因敲除小鼠尾静脉高压注射pAAV-HBV1.2，建立HBV感染动物模型，研究HBV感染状态下RORγt对相关炎性因子及HBV表达载量的影响，分析RORγt对HBV感染小鼠在IL-17基因缺失状态下肝脏炎症的直接调控机制；通过RORγt反义寡核苷酸干扰技术减少RORγt基因表达，研究HBV感染状态下RORγt对肝组织炎症反应的调节作用；通过RORγt与反应疾病严重程度的各临床指标的相关分析，阐明RORγt随疾病严重程度进展的变化规律，揭示RORγt在慢性HBV感染的表达变化。