矮地茶抗病毒和免疫调控协同治疗乙肝的药效物质基础及作用机制研究

Chronic hepatitis B is a widespread and great harmful refractory disease, there is still no ideal method and drug to cure it. Our previous studies showed that traditional Chinese medicine Ardisia japonica has significant curative effect on chronic hepatitis B. However, the chemical constituents of Ardisia japonica, the effective substances and the mechanism remain unclear. Based on the traditional Chinese medicine characteristics of multi-component, multi-target and overall synergy, we hypothesized that the anti-viral components of Ardisia japonica can directly inhibit hepatitis B virus (HBV) replication, meanwhile, indirectly remove HBV and repair liver injury by modulating the body immune system. This project intent to study the therapeutic substances of Ardisia japonica from two aspects of antivirus and immunomodulatory. We will isolate and identify the chemical constituents of Ardisia japonica by molecular imprinting, chromatographic and spectroscopic techniques. Screening and identifying the anti-HBV active components extracted from Ardisia japonica by HBV-expressing HepG2.2.15 cells, HBV transgenic mice and duck hepatitis B virus model. Investigating the effect of Ardisia japonica on endogenous metabolites in immunological hepatic injury rats by metabonomics, looking for potential biomarkers and related metabolic pathways, and elucidating the mechanism of Ardisia japonica antivirus and immunomodulatory in the synergistic treatment of chronic hepatitis B. This project provides an important scientific basis for the clinical application of Ardisia japonica, and also provide new ideas and methods for the research and development of anti-HBV drugs.

乙肝是流行广泛、危害极大的难治性疾病，目前尚无理想的根治方法和药物。前期研究发现中药矮地茶治疗乙肝效果显著，但其化学成分、药效物质与作用机理不明确。根据中药多成分、多靶点和整体协同作用的特点，我们推测：一方面矮地茶抗病毒成分直接抑制乙肝病毒（HBV）复制；另一方面，通过调动机体免疫系统间接清除HBV并修复肝损伤。本项目将从抗病毒和调免疫二个方面对矮地茶药效物质展开研究，采用色谱波谱、分子印迹等多种技术对矮地茶化学成分进行分离表征，通过HBV-DNA转染HepG2.2.15细胞、HBV转基因小鼠、鸭乙肝模型筛选矮地茶中抗HBV活性成分，运用代谢组学技术考察矮地茶对免疫性肝损伤大鼠内源性物质代谢的影响，寻找潜在生物标志物及其相关代谢通路，阐明矮地茶免疫调节和抗病毒协同治疗乙肝的作用机制。本项目将为矮地茶的临床应用提供重要科学依据，为抗乙肝药物的研究提供新的研究思路。

研究背景：乙肝是流行广泛、危害极大的难治性疾病，目前尚无理想的根治方法和药物。我们前期研究发现中药矮地茶对乙肝病毒（HBV）有显著抑制作用，同时也能调节机体免疫系统发挥抗炎保肝功效，但由于化学成分较复杂，药效物质不清楚，作用机理不明确，使其临床应用受到很大的限制。主要研究内容和结果：对矮地茶中主要的化学成分进行分离鉴定和定性定量分析检测，筛选出矮地茶中主要化学活性成分为岩白菜素、芦丁、山柰酚、槲皮素、没食子酸等；利用RAW264.7、LO2、HepG-2、免疫性肝损伤等细胞或动物模型，通过ALT、AST、ALP、SOD、MDA、HYP、GSH-Px、CD4、CD8等生化指标的测定，结果显示矮地茶具有体内外抗炎和保肝降酶、免疫调节等功效；利用网络药理、分子对接及代谢网络等技术从分子水平寻找抗炎保肝潜在生物标志物，结果表明矮地茶抗炎保肝作用可能跟AKT1、IL6、VEGFA、CASP3几个关键靶点相关。相关研究的详细数据以论文见刊或发表于后续的研究报道中。科学意义：以中医药整体观、辩证观为指导，结合现代分析、生物和药理等多学科实验方法和手段，揭示矮地茶通过多成分、多靶点共同作用的特点，具有一定的科学价值，将为矮地茶治疗肝炎的临床应用提供一定参考。