基于代谢组学及新生树鼩HB模型的丁香苦苷抗HBV作用机制研究

Hepatitis B (HB) is a highly infectious disease that seriously threatens human health, existing anti HBV drugs have many problems such as poor specificity and adverse reactions, finding effective anti HBV drugs is the key to solve this health problem. Syringopicroside is an antiviral component obtained from the Chinese medicinal lilac,the research group has confirmed that Syringopicroside has the characteristics of natural liver enrichment, with significant effect on HBV treatment, and no toxic side effects, and has been authorized the invention patent. The drug is expected to become a new anti HBV drug, but its mechanism is not clear, which limits the development of new drug. We select new-born tree shrews of the lower primate species that are closest to humans nature to construct an animal model of HB, we use urine, blood, and cell fluid metabolomics techniques to determine this by LC-MS/MS and PCMS,Under the premise of determining the effectiveness of the drug on the neonatal tree shrews HB model,to reveal the metabolized drug biomarkers in model animals, to search for differential markers, and track all the metabolic pathways affected by the drug, and the key metabolic pathways for the effect of Syringopicroside on HBV and the treatment of HB are screened out, to discover the key metabolism enzyme/protein information of HB and explain the biochemical mechanism of its effectiveness, so that it will lay a foundation for the subsequent research on the mechanism of action of Syringopicroside on the treatment of HB based on metabolic enzyme/protein target.

乙型肝炎（HB）是严重威胁人类健康的高发性传染病，现有抗乙型肝炎病毒 (HBV)药物存在专属性差、不良反应多问题，寻找高效抗HBV药物是解决这一健康问题的关键。丁香苦苷是从中药紫丁香中获得的抗病毒成分，课题组前期证实该成分具有天然靶向肝脏属性，体内外抗HBV效果显著，无毒副作用，已获授权发明专利，但其抗病毒作用机制尚不清楚，限制了该药物深度开发。本课题选用与人类亲缘关系最近的灵长类小动物新生树鼩构建HB动物模型，利用尿液、血液及细胞液代谢组学技术，应用LC-MS/MS和PCMS等分析手段，确定该药对新生树鼩HB模型有效的前提下，揭示该药在模型动物体内代谢生物标记物群，寻找差异标记物，追踪其影响的全部代谢通路，从中筛选出丁香苦苷对HBV及治疗HB作用的关键代谢通路，发现HB关键代谢酶/蛋白信息，阐释其有效性的生化机制，为后续基于代谢酶/蛋白作用靶点的丁香苦苷治疗HB的作用机制研究奠定基础。

乙型肝炎（HB）是严重威胁人类健康的高发性传染病，现有抗乙型肝炎病毒 (HBV)药物存在专属性差、不良反应多问题，寻找高效抗HBV药物是解决这一健康问题的关键。丁香苦苷是从中药紫丁香中获得的抗病毒成分，体内外抗HBV效果显著，无毒副作用，已获授权发明专利，但其抗病毒作用机制尚不清楚，限制了该药物深度开发。本项目选用树鼩和HBV转基因小鼠构建HB动物模型，完成了丁香苦苷对树鼩乙肝模型、HBV转基因小鼠模型及HepG2.2.15细胞模型的药效学验证，结果显现3种动物模型的HBeAg、HBsAg以及HBV-DNA含量明显降低，证实了丁香苦苷具有很强的抗HBV作用。在此基础上利用尿液、血液及细胞上清液代谢组学技术和肝脏蛋白质组学技术，应用LC-MS/MS和PCMS等分析手段，血清代谢组学筛选了42个与乙肝疾病模型相关的血清生物标记物，涉及的代谢通路有鞘脂代谢、甘油磷脂代谢、亚油酸代谢、脂肪酸降解等11个；尿液代谢组学结果确定了45个尿液组相关疾病生物标记物，涉及的代谢通路有苯丙氨酸代谢、戊糖和葡萄糖醛酸酯的相互转化、酪氨酸代谢、核黄素代谢等13个；细胞外液代谢组学结果显示丁香苦苷治疗后有77个代谢物含量向空白组方向回调，共涉及8条代谢通路。肝脏蛋白质组学研究结果显示，回调的HBV相关的蛋白有12个，上调的5个，下调的7个，涉及到的生物进程(BP)有175个，细胞组分(CC)22个，分子功能(MF)34个，共涉及49条通路。本研究发现了HB关键代谢酶/蛋白信息，为后续基于代谢酶/蛋白作用靶点的丁香苦苷治疗HB的作用机制研究奠定基础。本项目发表论文6篇，其中SCI论文3篇，核心期刊3篇，培养硕士研究生3名。