补肾方通过诱导中性粒细胞释放NETs活化浆细胞样树突状细胞抗HBV的机制研究

补肾方是治疗慢乙肝有效验方，RCT证实其可提高HBeAg和HBsAg阴转率，与改善浆细胞样树突细胞(pDC)功能有关。pDC表达内源性干扰素-a启动HBV特异性免疫应答是HBV清除的关键机制，ROS依赖的中性粒细胞胞外诱捕网(NETs)可激活此应答。我们发现，中性粒细胞miR-223抑制IL-6-p47phox通路减少ROS产生；补肾方可降低患者miR-223表达，促进NETs释放。提出假说：补肾方降低中性粒细胞miR-223表达，激活IL-6-p47phox-ROS通路，促进NETs释放以激活pDC产生干扰素-a启动免疫应答抗HBV。拟观察补肾方对HBV感染免疫耐受者中性粒细胞miR-223、ROS、NETs和pDC及干扰素-a表达的影响；体外探索HBV干预中性粒细胞和pDC机制及该方作用靶点；应用HBV持续感染小鼠模型，研究补肾方促进NETs释放激活pDC表达干扰素-a抗HBV的机制。

Bushen Recipe is an effective prescription for the treatment of chronic hepatitis B. In previous RCT researches, we found that the HBeAg seroconversion rate and HBsAg seroconversion rate were increased after patients were administrated with Bushen Recipe ,which is correlated with the improvement of pDCs. The key mechanism of HBV clearance is to activate HBV specific immune response by endogenous IFN-a expressed in pDC. ROS dependent extracellular traps (NETs) released by neutrophils can activate the expression of IFN-α in pDC. We found that miR-223 expressed by PMN inhibited ROS production through IL-6-p47phox pathway. Bushen Recipe can reduce the expression of miR-223 and promote the release of NETs. The hypothesis is put forward that Bushen Recipe treated HBV infection by initiating immune response, and the relevant mechanism is that Bushen Recipe reduced the expression of miR-223 in PMN and promoted the release of NETs by activating IL-6-p47phox-ROS pathway, and then the NETs activated pDC to produce IFN-α , which resulted in the restarting of immune response to HBV . The following experiments will be done. The effects of Bushen Recipe on the expression of miR-223, ROS, NETs, pDC and IFN-α in patients with chronic HBV infection in immunotolerant stage, and then the intervention of HBV protein on the function of neutrophils and PDC in vitro and the regulatory effect of the recipe will be explored. Finally, the mechanism of the formula blocking the chronicity of hepatitis B infection through promoting the release of NETs and activating pDC to produce IFN-α by using the mouse model of HBV persistent infection.