解毒凉血健脾方通过调控应急造血定向分化治疗急性肝衰竭的机制研究

The increase of neutrophil-lymphocyte ratio (NLR) in peripheral blood was demonstrated to reflect circular amplification of immune injury, which is associated with the prognosis of patients with acute liver failure. In our clinical studies, therapy of Jiedu-Liangxue-Jianpi Decoction was effective to improve survival of patients with acute liver failure by tempering NLR. Previous studies indicated that "the mobilization of granulocytes and lymphocytes exhaustion" might play a principal role in elevated peripheral NLR. However, it does not totally explain this phenomenon. In our previous work, the acute liver failure mouse model established with administration of ConA performed an increase of NLR. In addition, the number of hematopoietic stem and progenitor cells significant elevated, suggesting that hematopoiesis contributes to regulation of NLR. Accordingly, we proposed that Jiedu-Liangxue-Jianpi Decoction could treat acute liver failure by mediating lineage commitment of hematopoiesis and the consequent alteration of NLR. We intends to: 1) in individual level, comprehensively evaluate the efficacy of the decoction for liver damage and identify the major components for mediating NLR through ConA acute liver failure model; 2) in cytological level, explore the regulation of the drugs to the quantity and function of hematopoietic stem cells; 3) in molecular level, investigate the mechanism of the drugs on interfering with differentiation of hematopoietic stem cells. Clarify the key target of the decoction will shed lights on new therapeutic targets and strategies for treatment of liver failure.

外周血粒细胞淋巴细胞比例（NLR）升高反映了机体免疫损伤循环放大，与急性肝衰竭患者的病情和预后密切相关。我们临床研究发现：解毒凉血健脾方能够调节肝衰竭患者的NLR，改善生存率。既往研究认为"粒细胞动员和淋巴细胞耗竭"是导致外周血NLR升高的重要原因，但仍存在其他机制。我们前期应用ConA急性肝衰竭小鼠模型，发现NLR升高，同时造血干祖细胞数目增加，提示造血生成参与 NLR的调节。据此我们提出:"解毒凉血健脾方通过调控应急造血分化方向，影响NLR治疗急性肝衰竭"。本课题拟1）在整体水平采用ConA急性肝衰竭模型，验证全方及拆方的疗效，筛选出调节NLR治疗急性肝衰竭的关键药物；2）在细胞水平应用流式分选技术，明确药物对造血干祖细胞数量及功能的调节作用；3）在分子水平分析药物干预造血干祖细胞定向分化的机制，探讨解毒凉血健脾方及关键药物的作用靶点，为临床中医药治疗急性肝衰竭提供新的治疗策略和手段。

急性肝衰竭是由病毒、药物、毒物及酒精等多种因素引起的急性大面积肝细胞坏死 ,肝脏功能发生严重障碍。随着乙肝疫苗接种的普及以及抗病毒药物的应用,重症急性肝损伤的发病率逐年下降。但是由于缺乏阻断急性、大面积肝细胞坏死的有效药物,重症急性肝损伤的死亡率仍居高不下(70-80%)，我国每年因此而死亡的人数也高达10-2 0万。因此，研制靶点明确、有效的抗急性肝坏死药物是治疗急性肝衰竭、降低其高死亡率的关键。外周血粒细胞淋巴细胞比例(NLR)升高反映了机体免疫损伤循环放大，与急性肝衰竭患者的病情和预后密切相关。我们研究发现：造血干/祖细胞应急造血定向分化不是影响急性肝衰竭小鼠外周血NLR升高的主要原因，而粒细胞大量动员和淋巴细胞过度耗竭是肝脏损伤加重的可能机制。升麻是解毒凉血方调控造血发育治疗急性肝衰竭的关键药物，升麻能够通过抑制粒细胞向肝脏的动员和胸腺T淋巴细胞的凋亡，从而使降低外周NLR水平，抑制机体免疫损伤的循环放大，发挥肝保护作用。进一步阐明了中医药如何在治疗慢加急性肝衰竭中发挥作用。