基于模式生物斑马鱼的甘草对何首乌致特异质肝损伤的保护作用研究

Study on the protective effect of Glycyrrhiza uralensis Fisch.based on the model organism zebrafish on specific heterogeneous liver injury caused by Polygonum multiflorum Thunb.

目的:探讨甘草是否能够对何首乌致炎症特异质型肝损伤起到保护作用,从安全药理学层面阐明二者配伍的合理性与可行性。方法:采用脂多糖(LPS)联合何首乌提取物(PME)或大黄素(EMO)建立特异质肝损伤斑马鱼模型,并给予不同浓度的甘草提取物(GUE)或甘草组分异甘草素(ISL)/甘草次酸(GA),通过观察斑马鱼肝脏表型、中性粒和巨噬细胞的迁移行为,进行肝脏组织病理学分析,测定肝脏谷丙转氨酶(ALT)和谷草转氨酶(AST)活力,并考察FXR、SHP基因的mRNA水平变化,从提取物和单成分配伍两个层面探究甘草对何首乌致炎症特异质肝损伤的保护作用。结果:在炎症应激状态下,GUE、ISL和GA可显著抑制模型药物导致的斑马鱼肝脏变性、肝脏面积和明亮度总和的减小及ALT和AST活力升高,同时减轻特异质肝损伤相关的炎症反应,减少神经丘周围免疫细胞个数。不同浓度的甘草抑制了炎症应激状态下何首乌诱发的FXR、SHP基因mRNA水平下降。结论:GUE及主要成分对PME及主要成分致炎症应激状态斑马鱼肝损伤具有显著的保护作用,其保护作用机制可能与FXR-SHP轴有关。

Objective:Exploring whether Glycyrrhiza uralensis Fisch.can protect against inflammatory heterogeneous liver injury caused by Polygonum multiflorum Thunb.,and elucidating the rationality and feasibility of the compatibility of the two from the perspective of safety pharmacology.Methods:A specific heterogeneous liver injury model of zebrafish was established using lipopolysaccharide(LPS)combined with Polygonum multiflorum extract(PME)or emodin(EMO),and different concentrations of glycyrrhiza uralensis extract(GUE)or Glycyrrhiza uralensis Fisch.components isoglycyrrhetinic acid(ISL)/glycyrrhetinic acid(GA)were administered.The liver phenotype,neutrophil and macrophage migration behavior of zebrafish were observed for liver histopathological analysis,Measure the activity of liver alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and investigate the mRNA level changes of FXR and SHP genes.Explore the protective effect of Glycyrrhiza uralensis Fisch.on inflammatory specific liver injury caused by Polygonum multiflorum Thunb.from the perspectives of extract and single component compatibility.Results:Under inflammatory stress,GUE,ISL and GA significantly inhibited model drug-induced zebrafish liver degeneration,reduction of total liver area and brightness and elevation of ALT and AST viability,while attenuating inflammatory responses associated with idiosyncratic liver injury,and decreasing the number of immune cells around the neural thalamus.Different concentrations of Glycyrrhiza uralensis Fisch.inhibited the decrease in mRNA levels of FXR and SHP genes induced by Polygonum multiflorum Thunb.under inflammatory stress.Conclusion:GUE and its major components had significant protective effects against PME and its major components on zebrafish liver injury in the inflammatory stress state,and the mechanism of their protective effects might be related to the FXR-SHP axis.