基于PPARs-NF-κB信号传导通路探索瘀胆肝损伤机制及参芪扶正保护作用的研究

The mechanism of cholestatic liver injury remains unclear and lead to the difficulty of treatments. Our studies have demonstrated that PPARs-NF-κB signaling pathway may be the key regulatory factor in cholestatic liver injury, and astragalus mongholicus could ameliorate cholestatic liver injury, improve the inflammatory environment of liver, but the exact mechanism is obscure. Although Bone marrow-derived mesenchymal stem cells (BMSCs) and Shenqi Fuzheng injection (SFI) plays an important effect in the regulation of NF-κB in treatment of cholestatic liver injury, what is the role of PPARs-NF-κB signaling pathway in the treatment? So the purpose of this study is to investigate the role of PPARs-NF-κB signaling pathway in cholestatic liver injury and protective effect of BMSCs combine with SFI in treatment. On the basis of the in vivo and in vitro experiments, we will investigate the regulative mechanism of PPARs and NF-κB in cholestatic liver injury and treatment of BMSCs combine with SFI, assess the efficacy of SFI and BMSCs, explore the reasonable therapeutic principle for cholestatic liver injury, and search for novel regulatory factors. Moreover, we will also study that whether or not SFI could promote proliferation, migration, differentiation, and improve the low implantation rate and limited conversion rate of BMSCs. Our research will offer the experimental and theoretical basis for the mechanism of cholestatic liver injury and therapy of BMSCs combine with SFI, and also create a new therapeutic approach for cholestatic liver disease.

瘀胆肝损伤机制目前尚未完全阐明，临床缺乏有效的治疗手段。本课题组研究证实PPARs-NF-κB信号通路在瘀胆肝损伤中起着关键的调节作用，黄芪对瘀胆肝具有保护作用。但这一信号传导通路在瘀胆肝损伤中具体调控机制尚不清楚，骨髓间充质干细胞（BMSCs）及参芪扶正注射液（SFI）虽能够作用于NF-κB，但能否通过调控PPARs-NF-κB信号通路发挥治疗作用？本课题拟在瘀胆肝大鼠模型及体外实验中深入研究PPARs-NF-κB信号通路在瘀胆肝损伤中的作用，明确这一信号通路在瘀胆肝损伤中与氧化、炎症、凋亡、自噬及纤维化间的关系及调控机制，探讨SFI联合BMSCs治疗瘀胆肝损伤的效果及机制，并明确SFI能否通过提高BMSCs定植率、转化率及促进其向肝细胞分化，而增强其治疗效果。课题将为瘀胆肝损伤机制及中药结合BMSCs移植治疗瘀胆肝损伤提供实验及理论依据，也将为瘀胆肝这一临床难题提供新的治疗途径。

瘀胆肝是由于肝细胞和（或）胆管细胞水平上的胆汁形成或流动的损害，导致胆汁淤积于肝脏，引起肝损伤、肝硬化、甚至肝衰竭。瘀胆肝是临床病人死亡率较高的直接原因，临床缺乏特效治疗药物及手段，因此明确其具体机制，寻找有效的治疗手段是一个亟待解决的临床难题。我们的研究表明：1.通过体内建立瘀胆肝大鼠模型，体外采用胆汁或胆汁化血清造成肝细胞损伤，证实了标本上清成分中PPARs、NF-κB及氧化、炎症因子等水平的变化。组织学观察肝细胞形态、超微结构变化，蛋白及基因水平检测PPARs-NF-κB信号通路及氧化、炎症、凋亡、自噬及纤维化特征因子的表达，明确了PPARs-NF-κB这一信号通路在瘀胆肝损伤中的调控机制；2.利用参芪扶正液（SFI）进行干预后，血清中ALT、AST的水平明显下降，同时检测到肝脏组织中IL-6、TNFα的水平，减轻了炎症因子浸润及对肝脏损伤的程度；3.SFI参芪扶正注射液能减轻大鼠早期梗阻性黄疸肝脏损伤，可能通过调节 PPAR-γ 蛋白，抗炎、抗氧化应激减轻梗阻性黄疸大鼠肝损伤；4.穿心莲内酯对ANIT所致的大鼠肝损伤具有保护作用，通过抗炎和抗氧化机制，减轻胆汁淤积程度。本课题的研究明确了胆汁淤积发生及发展的具体机制，为瘀胆肝损伤提供实验及理论依据，证实了中药对瘀胆肝治疗的效果及机制，也将为瘀胆肝的治疗方案提供新理论依据及新思路。