基于细胞外组蛋白-一种新的炎症分子探讨清肠利肝方治疗肝衰竭的作用机制

Liver failure is an acute severe syndrome with high mortality rate. Excessive and long-lasting inflammatory response as the second hit to liver leads to liver failure. Extracellular histones are the newly discovered endogenous damage-associated molecular patterns(DAMPs) and the important inflammatory mediators in the pathogenesis of liver failure by activating the inflammatory signals via Toll-like receptors. Qing chang li gan formula (QCLGF) is a traditional Chinese medicine (TCM) formula with function of qi-boosting, toxin-resolving and promoting blood circulation developed on the basis of Dachengqitang, a very famous traditional formula. QCLGF has been used in our hospital for a long time and is proved to be effective for patients with liver failure. However, it is still lack of data revealing the mechanism of QCLGF for treating liver failure based on the modern techniques. This research tries to take extracellular histones as the starting point to study the mechanism of QCLGF for liver failure. Through concanavalin A (ConA) or acetaminophen (APAP) induced mouse model with acute liver failure and macrophage cell lines developed from mice abdominal cavity, we will observe the effect of QCLGF on the activation of macrophage mediated by extracellular histones and on the inflammatory signaling pathway. A randomized controlled clinical trial with hepatitis B patients with acute on chronic liver failure will be performed to examine the levels of extracellular histones and the extent of activation of peripheral blood mononuclear macrophage with or without QCLGF treatment. The results from the study will help to clarify the molecular mechanism of QCLGF for treating liver failure and promote the popularization and application of TCM.

肝衰竭是临床常见的危急重症，死亡率极高。过度持久的炎症反应对肝脏的二次打击是导致肝衰竭的重要原因，细胞外组蛋白是新发现的一种内源性损伤相关分子模式（DAMPs）分子，通过Toll样受体（TLRs）活化炎症信号，是导致肝衰竭的重要炎症介质。"清肠利肝方"（QCLGF）是在小承气汤基础上创立的解毒、活血、益气方剂，作为北京佑安医院院内协定处方治疗肝衰竭取得了良好的临床疗效，但作用机制缺乏现代医学基础研究资料支持。本课题以细胞外组蛋白为切入点，通过以刀豆蛋白A（ConA）及对乙酰氨基酚（APAP）肝衰竭小鼠模型为研究对象的体内研究，以小鼠腹腔巨噬细胞系为研究对象的体外研究，以乙肝慢加急性肝衰竭患者为研究对象的临床研究，采用现代医学研究技术，探索QCLGF对细胞外组蛋白水平、其介导的巨噬细胞活化及炎症信号通路的影响，从分子水平揭示QCLGF治疗肝衰竭的作用机制，促进QCLGF的推广应用。

肝衰竭是临床常见的危急重症，死亡率极高。过度持久的炎症反应对肝脏的二次打击是导致肝衰竭的重要原因；细胞外组蛋白是新发现的一种内源性损伤相关分子模式（DAMP）分子，通过Toll 样受体（TLRs）或炎症小体等途径活化炎症信号，是导致肝衰竭的重要炎症介质。“清肠利肝方”（QCLGF）是在小承气汤基础上创立的解毒、活血、益气方剂；过去十多年来清肠利肝方作为首都医科大学附属北京佑安医院的院内协定处方治疗肝衰竭已经取得了良好的临床疗效，但其作用机制尚缺乏现代医学基础研究资料的支持。本项课题以细胞外组蛋白为视角，通过制备不同病因导致的小鼠肝衰竭模型进行体内研究，同时以小鼠巨噬细胞系为研究对象进行体外研究，并结合乙肝导致的慢加急性肝衰竭患者为研究对象的临床研究，采用现代医学研究技术，探索清肠利肝方对细胞外组蛋白的水平以及其介导的巨噬细胞活化及炎症信号通路的影响等。本课题组历经4年研究，通过动物、细胞和临床三个层面的工作，清楚阐明了清肠利肝方在治疗肝衰竭中确具重要作用，其作用机制可能与降低细胞外组蛋白水平有关；即清肠利肝方通过抑制细胞外组蛋白介导的炎症损伤反应发挥治疗肝衰竭的作用。我们的课题研究不仅从基础科研的角度验证了清肠利肝方的疗效，而且通过对其作用机制的深入探讨和挖掘，为清肠利肝方的推广应用提供了重要的科学线索，并对其他类似病理生理的疾病亦有重要的应用潜力。