基于代谢组学经时谱效关系融合网络药理学的血必净抗脓毒症药效物质发现及分子机制研究

Xuebijing (XBJ) injection is the only new Chinese medcine approved by China Food and Drug Administration (CFDA) for the treatment of sepsis and the associated multiple organ dysfunction syndrome. Although the efficacy of XBJ has been supported by clinical practice and experimental data, its bioactive components and mechanism of action remain largely unknown. Therefore，it is difficult to ensure the safety and rationality in clinic and it often cause adverse drug reactions. In our previous work, high resolution mass spectrometry was applied to analyze complex ingredients of XBJ injection and 162 components were identified. In this research, we will focus on the in vivo process of multi-component XBJ injection. The correlation between characterization spectrum of exogenous compounds in vivo and global efficacy reflected by endogenous metabolites will be studied with integration of metabolomics and bioinformatics. The components with strong correlation with endogenous biomarkers are considered as potential effective constituents of XBJ injection. In addition, the targets of these effective constituents will be further explored based on the network pharmacology and molecular docking technology, and it is aimed to establish a relationship network of components, targets and the disease. The potential effective components, related targets and metabolic pathways will be verified by subsequent biological experiments. Finally, the study is expected to elucidate the underlying mechanism that how the synergic essence of XBJ achieve therapeutic efficacy via multiple components interacting with multiple cellular targets. The results of this research will reveal the scientific connotation of XBJ antisepsis efficacy, and the new research thought will improve efficiency to discover the bioactive components of Chinese herbal formula and the underlying active mechanisms.

血必净注射液是唯一经CFDA批准的用于治疗脓毒症的二类新药，临床疗效确切。然而，药效物质不清和作用机制不详严重影响了其临床使用的安全合理，导致不良反应频发。在前期研究中，我们基于高分辨质谱检测建立了血必净体外复杂成分解析策略，基本明确了其化学组成。本项目拟将前期体外研究深入到体内过程，通过代谢组学技术和生物信息学手段将给药后体内原型和代谢产物经时变化的“谱”与内源性代谢标志物整体动态变化的“效”关联分析，筛选相关系数较大的成分作为血必净抗脓毒症的潜在药效物质；利用网络药理学和反向分子对接技术寻找这些药效成分的作用靶点，全面构建“活性成分群-作用靶点-疾病”网络，进一步采用分子生物学手段对潜在药效成分（群）及作用靶点和通路进行验证，阐释血必净抗脓毒症多成分、多靶点的整体作用机制。研究成果将为揭示血必净抗脓毒症的科学内涵提供理论依据，研究思路将为高效研究中药复方药效物质和作用机制提供方法借鉴。

血必净注射液临床应用广泛且治疗脓毒症疗效确切，然而其药效物质基础及作用机制尚不明确。本项目围绕“哪些成分有效”及“如何起效”的科学问题，对血必净注射液开展了系统研究。化学成分研究结果显示，在血必净中共鉴定得到162种化合物，在大鼠血浆、尿液和粪便样本中共鉴定得到102种化合物，包括26种原型成分和76种代谢物，同时描绘了血必净主要成分在大鼠体内的药代动力学过程和组织分布情况，基本明确了其体内外化学物质组成。药效学结果表明，血必净能够显著改善脓毒症大鼠行为学、提高大鼠生存率、改善组织损伤、抑制炎症因子表达。代谢组学研究结果表明，血必净干预后，脓毒症大鼠血浆中有32个代谢物得到显著回调，主要涉及牛磺酸和亚牛磺酸代谢，缬氨酸、亮氨酸和异亮氨酸生物合成，烟碱和烟酰胺代谢，鞘脂代谢等；肺组织中有15种代谢物表现出显著回调趋势，涉及谷氨酰胺和谷氨酸代谢、丙氨酸、天冬氨酸和谷氨酰胺代谢及鞘脂代谢等。进一步，以内源性代谢物变化来反映整体药“效”，并结合血必净化学成分“谱”，通过采用数据挖掘技术和网络药理学研究方法，整合关联分析全面构建血必净“活性成分群-作用靶点-疾病”网络，获得血必净抗脓毒症肺损伤的关键核心靶点37个。最后，基于建立的多成分多靶点系统网络药理模型，采用细胞分子生物学技术，在体内组织水平和体外细胞水平上对血必净4个潜在活性成分开展效应成分（群）的关键靶点及通路验证，结果表明，芍药苷、氧化芍药苷、羟基红花黄色素A和洋川芎内酯I能够调控PI3K/Akt/mTOR信号通路改善炎症反应，减轻脓毒症肺损伤。以上研究不仅揭示了血必净的关键药效物质及作用机制，为其抗脓毒症的科学内涵提供了理论依据，同时也为中药复方药效物质基础及作用机制研究探索了新方法。研究相关内容已发表学术论文3篇，其中SCI文章2篇，另有2篇SCI及2篇中文核心正在投稿中；获得科研奖励2项；参加国内外学术会议3次，举办学术交流会议6次；培养博士后1名，硕士研究生3名。