基于NMR、LC-MS技术探讨解毒化瘀颗粒拮抗急性肝衰竭大鼠的代谢组学研究

Acute hepatic failure is one of the clinical emergent and severe cases, its pathogenesis is not yet entirely clear, which mainly tends to be the result of comprehensive factors. It determines treating acute hepatic failure should be considered from the whole and this is identical with the holistic concept of Traditional Chinese Medicine (TCM) perfectly. Based on previous studies, the rat model with acute hepatic failure in type of cementing toxin and stasis copied by D - GalN + LPS was made. Changes of metabolites in urine and serum in the process of making mold and intervention of formulas were observed dynamically through using the method of NMR and LC – MS. The rat model with acute hepatic failure and fingerprint of metabolites were built up. This fingerprint was record before and after giving the formula of cleaning toxin and stasis, its separated formulas including formula of cleaning toxin and formula of cleaning stasis. The information of iconic biomarkers which were closely related to acute hepatic failure and invention of compounds of Chinese herbs, were analyzed and identified by recognition technologies like PCA. And then, these key characteristic metabolites were further isolated, identified in structure and classified to be researched. The completion of project would help elucidate the pathogenesis of acute hepatic failure systematically, reveal the biological essence in the type of cementing toxin and stasis in this disease and make clear effectors, action pathway and targets of granules for cleaning toxin and stasis on antagonizing the acute hepatic failure. It would have an innovative significance on enriching theories of TCM effective carriers and syndrome differentiation.

急性肝衰竭是临床危急重症之一，其发病机制至今尚未完全明晰，主要倾向于由多种因素综合作用的结果，就决定了本病的治疗应从整体出发，这与中医学“整体观念”十分吻合。本课题组在前期研究基础上，以D-GalN+LPS复制“毒瘀胶结型”急性肝衰竭大鼠模型，运用NMR、LC-MS等方法，动态观测各组大鼠在造模和方药干预过程中尿液、血清中代谢物质的变化，构建急性肝衰竭大鼠模型和解毒化瘀方及其拆方（解毒方、化瘀方）给药前后的代谢物指纹图谱，应用PCA等识别技术，解析、辨识与急性肝衰竭发病及中药复方干预密切相关的标志性生物标记物所蕴含的信息，然后进一步对这些关键性特征代谢物行分离、结构鉴定及分类研究。本项目的完成有助于从系统层面阐明急性肝衰竭的发病机制，揭示本病“毒瘀胶结”证的生物学物质实质，明确解毒化瘀颗粒拮抗本病的体内效应物及作用途径、靶点。它对丰富中医疗效载体——中医证型理论具有创新性的意义。

急性肝衰竭是临床危急重症之一，其发病机制至今尚未完全明晰，主要倾向于由多种因素综合作用的结果，就决定了本病的治疗应从整体出发，这与中医学“整体观念”十分吻合。本课题组在前期研究基础上，以D-GalN+LPS复制“毒瘀胶结型”急性肝衰竭大鼠模型，运用NMR、LC-MS等方法，动态观测各组大鼠在造模和方药干预过程中尿液、血清中代谢物质的变化，构建急性肝衰竭大鼠模型和解毒化瘀方及其拆方（解毒方、化瘀方）给药前后的代谢物指纹图谱，应用PCA等识别技术，解析、辨识与急性肝衰竭发病及中药复方干预密切相关的标志性生物标记物所蕴含的信息，然后进一步对这些关键性特征代谢物行分离、结构鉴定及分类研究。本项目的完成有助于从系统层面阐明急性肝衰竭的发病机制，揭示本病“毒瘀胶结”证的生物学物质实质，明确解毒化瘀颗粒拮抗本病的体内效应物及作用途径、靶点。它对丰富中医疗效载体——中医证型理论具有创新性的意义。