补肾方调节MAVS介导的信号通路发挥抗炎、抗病毒作用机制及其主体疗效中药组

The primary of tonifying kidney, the secondary of clarity and removability is an effective prescription treatment of tonifying kidney to treat chronic hepatitis B, with regulating immunity, suppression of HBV replication and improve inflammation. Mitochondrial antiviral signaling protein (MAVS) is the key one of regulating innate immune signaling pathways that not only stimulates the interferon pathway named activation of type 1 such as TBK1, viperin and so on to inhibite the viral replication, but also to stimulate the prescription of tonifying kidney downstream's activation of NF-κB pathway such as TRAF6, TRADD to promote inflammation. Preliminary studies confirmed that the prescription of tonifying kidney may raise MAVS.Thus we proposed a hypothesis that the prescription of tonifying kidney plays an antiinflammatory and antiviral part by regulating MAVS-mediated signaling pathway .This project uses immune liver injury model of HBV transgenic mice to study the prescription of tonifying kidney and its unsealed prescription of tonifying kidney and clarity and removability to promote the generation of interferon and the inhibition of viral replication through phosphorylation and ubiquitin-like activation of TBK1 complexion namely IKK,TBK1,DDX3,STING and to reduce inflammation through inhibition of FADD compound activation namely TRAF6,FADD,TRADD,RIP1 which clears the respective roles of the prescription of tonifying kidney that provides immunological basisfor the treatment based on the prescription of tonifying kidney.

"补肾为主，清化为辅"的补肾方是治疗慢乙肝的有效方剂，具有调节免疫、抑制乙肝病毒复制、改善炎症的作用。线粒体抗病毒信号蛋白(MAVS)是调节先天免疫信号通路的关键蛋白，可刺激TBK1、viperin等活化Ⅰ型干扰素通路抑制病毒复制，又可刺激下游的TRAF6、TRADD等活化NF-κB 通路促进炎症。前期研究证实，补肾方可以上调MAVS，由此提出假说"补肾方调节MAVS介导的信号通路发挥抗病毒、抗炎作用"。本项目采用HBV转基因小鼠免疫性肝损伤模型，研究补肾方及拆方(补肾、清化)通过活化TBK1复合体（IKK、TBK1、DDX3、STING）促进干扰素生成抑制病毒复制；通过抑制FADD复合体（TRAF6、FADD、TRADD、RIP1）活化减轻炎症。以明确补肾方组方各自作用环节，为补肾方治疗慢乙肝的疗效提供免疫学依据。

"补肾为主，清化为辅”的补肾方是治疗慢乙肝的有效方剂，具有调节免疫、 抑制乙肝病毒复制、改善炎症的作用。线粒体抗病毒信号蛋白(MAVS)是调节先天免疫信号通 路的关键蛋白，可刺激 TBK1、viperin 等活化I型干扰素通路抑制病毒复制，又可刺激下游 的 TRAF6、TRADD 等活化 NF-κB 通路促进炎症。前期研究证实，补肾方可以上调 MAVS，由此提出假说“补肾方调节 MAVS 介导的信号通路发挥抗病毒、抗炎作用”。本项目采用 HBV 转 基因小鼠免疫性肝损伤模型，研究补肾方通过活化 TBK1 复合体促进干扰素生成抑制病毒复制;通过抑制 FADD 复合体活化减轻炎症。研究结果表明：补肾方可显著降低ConA诱导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，改善肝组织纤维化程度，上调肝组织内CD68+细胞的表达；可显著降低小鼠血清HBsAg水平,降低肝组织内HBcAg的表达；可提高小鼠外周血清IFN-alpha及IFN-β水平,上调IRF-3、TBK-1、Viperin、STING、cGAS、WDR5、TAK-1等蛋白的表达,下调TRADD的表达，对pTAK-1（t187）及FADD（s194）、FADD（s191）表达无影响。在WDR5基因敲除慢乙肝感染小鼠模型中，补肾方也可改善肝组织损伤，下调外周HBsAg的表达，上调WDR5、sting、cGAS等蛋白的表达。提示补肾方具有减轻肝组织炎症、抗HBV作用,其作用机制与MVAS介导的信号通路有关。