补肾方通过DCs的干扰素诱导蛋白16信号通路发挥抗病毒和免疫调节作用机制研究

The DNA sensor, interferon-inducible protein 16(IFI16) in the nuclear of Dendritic cells (DCs) can identify the virus dsDNA, active the IFI16 - cGAS - STING (IFI16 pathway) and increase expression of type I interferon IL - 12, promote Th1 and Tc1 polarization. Early research has shown that Bushe recipe can improve DCs function in patients with chronic hepatitis b, promote Th1 and Tc1 polarization, suppress HBV replication and improve the result the expression of IFI16 in DCs ,which is transfection with HBV DNA. So there is a hypothesis that Bushe recipe may activate IFI16 pathway to improve the function of DCs and then promote Th1 and Tc1 polarization ,playing a role of HBV resistance. In this study, it is observed that the expression of IFI16 ,cGAS, STING, type I interferon, IL - 12 ,the ratio of Th1/2, Tc1/2 in liver of HBV transgenic mices and the intervention role of Bushe recipe. Then, SiRNA will be used to interfering DCs’cGAS, STING gene, which will co-culture with initial T cell to discusses how Bushe recipe DCs works on IFI16 pathways to mediate secretion of type I interferon, IL - 12 and the polarization of Th1, Tc1 . Based on the above research, it will clarify the immunological mechanisms of Bushe recipe in treatment of chronic hepatitis b.

树突状细胞（DCs）核内的DNA识别受体干扰素诱导蛋白16(IFI16)识别病毒dsDNA，激活IFI16-cGAS-STING通路（IFI16通路）上调I型干扰素、IL-12表达，促进Th1和Tc1极化。前期研究表明，补肾方可改善慢乙肝患者DCs功能，促进Th1和Tc1极化，抑制HBV复制；补肾方可改善HBV DNA转染导致的DCs内IFI16低表达。提出假说“补肾方活化IFI16通路改善DCs功能促进Th1和Tc1极化以发挥抗HBV作用”。本研究拟：观察HBV转基因小鼠肝内DCs的 IFI16、cGAS、STING 、I型干扰素、IL-12表达，肝内Th1/2、Tc1/2比率及补肾方的干预作用；siRNA干扰DCs的cGAS、STING基因，与初始T细胞共培养，探讨补肾方对DCs的IFI16通路介导I型干扰素、IL-12分泌和Th1、Tc1极化的影响。明确补肾方治疗慢乙肝免疫学机制。

树突状细胞(DCs)核内的DNA识别受体干扰素诱导蛋白16(IFI16)识别病毒dsDNA，激活IFI16-cGAS-STING通路(IFI16通路)上调I型干扰素、IL-12表达，促进Th1和Tc1极化。本课题研究内容为证实补肾方可通过活化 IFI16 介导的信号通路，改善DCs功能，促进I-IFN的表达，可通过旁路活化途径促进IL-12等细胞因子的表达，促进T细胞向 Th1和Tc1极化以发挥抗HBV作用。研究结果证实：补肾方可以显著改善ConA诱导的HBV转基因小鼠血清ALT、AST水平、减轻肝组织内炎性细胞的浸润，改善肝细胞的变性坏死，促进肝组织内CD86+细胞的增值；可显著降低小鼠血清 HBsAg 水平,降低肝组织内 HBcAg 的表达;可以上调小鼠血清中IFN-β、IFI-γ、IL-4、IL-12的表达水平，上调小鼠肝内IFI16、IRF3、STING、TBK-1蛋白的表达。在WDR5基因敲除慢乙肝转染小鼠模型中，补肾方可抑制rcccDNA小鼠外周血HBsAg的表达、改善肝组织内炎症损伤，上调肝脏内及DCs的IFI16、IFR3、STING等蛋白的表达；通过激活DCs内IFI16信号通路，从而进一步发挥APC作用，减缓T细胞活化时间，并促进T细胞极化，发挥抗病毒作用。