IL-27调控Th17细胞分化抑制原发性胆汁性肝硬化进展的研究

Primary biliary cirrhosis (PBC) is a chronic intrahepatic cholestatic autoimmune liver desease that can evolve into hepatic fibrosis, hepatic cirrhosis and hepatic failure. But the progression of PBC is heterogeneous. Nowadays,there is no good method to monitor disease progression. Our previous studies demonstrates that Th17 cells are the key immunal effector cells of PBC . The increased Th17 cells can lead to the occurrence of disease and liver fibrosis. IL-27 can inhibited fibrosis disease through regulating the Th17 cells differentiation. In this research, we use flow cytometric analysis to examined the lever of IL-27、 Th17 cells and related cytokines in peripheral blood and hepatic tissue.To clarify the expression,location,cell origin of IL-27 in PBC, we perform Immunohistochemical staining and RT-PCR.To observe the role of the IL-27 regulating Th17 differentiation and secretory function,spleen lymphocytes will be co-cultured with IL-27and anti-IL-27 in vitro. We use Western Blotting to explore the role of JAK / STAT signaling pathway in IL -27 regulating function. To further investigate the role of IL-27 in the pathological processes of PBC,the correlation between IL-27 expression and disease activity, including biologychemystry and Mayo score will be analyzed. The research will further elucidate the molecular mechanism of IL-27 inhibit liver fibrosis in PBC and provide a new biomarker and target for monitoring、prevention and treatment of PBC.

原发性胆汁性肝硬化(PBC)是一种胆汁淤积性自身免疫性肝病，易渐进发展为肝纤维化、肝硬化和肝衰竭。PBC进展异质性大，病情监测困难。我们前期研究证实PBC自身免疫反应主要由Th17细胞介导，IL-27是Th17细胞的主要调控因子。研究肺纤维化小鼠模型证实IL-27可以抑制成纤维细胞功能。为进一步阐明IL-27对Th17调控在PBC进程中的作用和机制，本项目拟检测PBC患者和小鼠外周血/肝中IL-27、Th17细胞、相关细胞因子水平以及IL-27、IL-27受体在肝组织的定位及表达，观察IL-27、IL-27抗体干预前后对Th17细胞分化及JAK/STAT、TGF-β/Smad信号通路的影响，从炎症和免疫调节的角度分析IL-27调控Th17与临床指标、病理分期及疗效的关系，阐明IL-27调节PBC免疫功能失衡抑制PBC进程的分子机制，为PBC病情评估、病程监测及疗效监控发现潜在的分子标志物。

原发性胆汁性胆管炎（PBC）又称原发性胆汁性肝硬化，是器官特异性的慢性胆汁淤积性自身免疫性肝病。本课题组前期研究发现在PBC中IL-27 p28亚基和IL-27受体WSX-1S和gp130亚基以及IL-17、IL-6、IL-20、IL-22、IL-23、TNF-α表达提高。推测IL-27水平可能会影响PBC进程。研究发现外源升高或降低IL-27都对PBC各个阶段肝病理无影响，p28、WSX-1S和gp130亚基无显著影响，但是它们的表达会随着PBC发展呈上升趋势。随着PBC的发展炎症因子分泌水平逐渐升高，注射IL-27后炎症因子表达水平较PBC组升高，但是降低IL-27后抑制炎症因子表达，但仍高于正常组。IL-27增加肝组织TH17细胞分化，Anti-IL-27处理降低肝组织中TH17细胞分化。L-27对Th17细胞中JAK2/STAT信号通路的影响较小，但与IL-27处理相比，Anti-IL-27处理显著抑制JAK2/STAT信号通路。IL-27和Anti-IL-27对PBC发展过程中SOCS1表达无影响，但Anti-IL-27缓解PBC发展过程中SOCS3表达的增加。IL-27和Anti-IL-27对PBC发展过程中Smad和p-Smad表达无显著影响，但相比IL-27，Anti-IL-27轻微缓解PBC发展过程中Smad和p-Smad表达。IL-27处理增加TGF-ꞵR2和TGF-ꞵR3表达。结果表明，外源升高或降低IL-27都对PBC各个阶段肝病理无影响，但是对炎症因子、Th17、JAK2/STAT和TGFβ/Smad信号通路有影响。