朝鲜大蓟中紫杉叶素基于HMGB1-NLRP3炎症小体信号轴调控酒精性脂肪性肝炎中炎性损伤的机制研究

Alcoholic steatohepatitis (ASH) is liver-damaging disease due to long-term overdrinking accompanied by inflammation, and no effective drug has been found yet. In the microenvironment of ASH, hepatocytes-Kupffer cells interaction system is presented with hepatocytes inflammatory injuries as a core part, activated Kupffer cells releasing inflammatory mediators. HMGB1 acts as an alarmin when hepatocytes inflammatory injuries occur, linking hepatocytes and Kupffer cells, based on the crosstalk mediated by NLRP3 between them. It is proposed that HMGB1-NLRP3 axis is the key signaling axis that can modulate ASH, on account of TLR and P2X7R experiment. Cirsium japonicum var. ussuriense (Regel) Kitam. (CK) is one of the most important medicines for Taiyin people according to the Chinese Korean medicine theory, for the treatment of liver disease characterized by “turbid blood and astringent Qi”. Previous research found that, taxifolin, content in the total flavonoids of CK, can ameliorate alcoholic streatosis, and inhibit the release of HMGB1. Base on this point, this research will combine In vitro and in vivo experiments, employ gene silencing technology. Regarding HMGB1 as the cutting point, in order to demonstrate the mechanism that taxifolin can modulate the activation of NLRP3 inflammasome by TLR4/RAGE and P2x7R signaling pathways, providing new targets and theoretical basis for the protection of ASH for Chinese Korean medicine.

酒精性脂肪性肝炎（ASH）是长期过量饮酒所致伴随炎症的肝脏损害性病变，目前尚无特效治疗药物。ASH的肝脏微环境中，以肝细胞炎性损伤为中心环节，被激活枯否细胞释放炎性介质，构成肝细胞-枯否细胞相互作用体系。HMGB1作为肝细胞发生炎性损伤的警报素，链接肝细胞与枯否细胞，基于NLRP3介导肝细胞和枯否细胞间交叉对话。由此提出，HMGB1-NLRP3是调控ASH的关键信号轴，基于TLR4和P2X7R实现。朝鲜大蓟属于朝医太阴人要药，常用于“血浊气涩“的肝脏疾病治疗。前期研究发现，朝鲜大蓟总黄酮中紫杉叶素（taxifolin）改善酒精性脂肪肝，抑制HMGB1释放。基于此，本研究拟结合动物与细胞实验，采用基因沉默技术，以HMGB1为切入点，明确紫杉叶素基于TLR4/RAGE与P2x7R信号通路协同调控ASH中NLRP3炎症小体活化的作用机制，为朝药防治ASH提供新靶标与理论依据。

酒精性脂肪性肝炎是长期过量饮酒所致伴随炎症的肝脏损害性病变，目前尚无特效治疗药物。酒精性脂肪性肝炎的肝脏微环境中，以肝细胞炎性损伤为中心环节，被激活巨噬细胞释放炎性介质。HMGB1作为肝细胞发生炎性损伤的警报素，链接肝细胞与巨噬细胞，基于TLR4和P2X7R实现HMGB1-NLRP3调控酒精性脂肪性肝炎。本课题通过酒精性脂肪肝动物模型与高脂饲料联合酒精模型以及肝损伤模型，从体内角度阐明朝鲜大蓟中紫杉叶素（taxifolin）基于HMGB1-P2X7R介导的NLRP3炎症小体的活化细胞特异性的调控脂肪蓄积与炎症，从而改善酒精性脂肪肝。另外，采用肝细胞或巨噬细胞，介导HMGB1-NLRP3信号轴，明确紫杉叶素干预酒精导致的肝细胞脂质稳态失衡与炎性坏死，进一步阐明HMGB1-TLR4-P2X7R是紫杉叶素调控ALD脂肪蓄积中关键靶细胞炎性关键信号轴，采用基因特异性沉默技术，阐明HMGB1-TLR4/P2X7R是紫杉叶素基于NLRP3炎症小体调节ASH中肝细胞-巨噬细胞之间炎性信号回路的关键上游受体，为紫杉叶素基于HMGB1介导的肝细胞-巨噬细胞交互对话调控ASH提供扎实的数据支撑，为朝药防治ASH提供新靶标与理论依据。