Efficacy And Mechanism Of Dipotassium Glycyrrhizinate-luteolin Premicelles In The Treatment Of Liver Injury

Acetaminophen was the most commonly used antipyretic and analgesic in clinical practice.The severe drug-induced liver injury was induced by excessive dosages of it.And only one antidote,N-acetylcysteine,was currently approved for clinical use.Moreover,this antidote had low bioavailability and was only effective in the early stages of liver injury.Therefore,it was of great clinical significance to study a safer and more effective therapeutic drug.The aim of this study was to establish glycyrrhizate-luteolin premicelles,evaluate their absorption in vivo and explore the pharmacological effects of dipotassium glycyrrhizate-luteolin premicelles on drug-induced liver injury in mice,as well as the influence of the high mobility group B1（HMGB1）and receptor of advanced glycation endproducts（RAGE）in the liver’s expression.Luteolin（LUT）based on dipotassium glycyrrhizate（DG）premicelles were established with solvent evaporation method,and the formulation was optimized with the encapsulation rate as an index.And its solid and solution were characterized and then the storage stability was evaluated;furthermore the chicken embryo allantoic membrane test and hemolysis test were used to evaluate the safety of LUT-DG.The average particle size,polydispersity index and Zeta potential of LUT-DG were 30.32±0.12 nm,0.138±0.02 and-27.0±0.51 m V respectively.The encapsulation efficiency of LUT-DG was 99.16±0.90 %.In the safety experiments,LUT-DG was no hemolysis and no bleeding in allantoic membrane test of chicken embryo.Dialysis bag method was used to evaluate the in vitro release of LUT-DG,The drug concentration in plasma and tissue after oral administration of LUT-DG in rats were determined by high-performance liquid chromatography（HPLC）,the pharmacokinetic parameters were calculated by DAS software version 2.1.1.Then evaluation of the oral bioavailability of LUT-DG.The solubility of LUT-DG was 12304.36 times that of LUT,and at 24 hours,the release amount of LUT-DG was 79.75±5.21 %;compared with LUT,the relative oral bioavailability of LUT-DG reached 171.72 %;the absorption of LUTDG at each time point in each tissue was higher than LUT.The mice liver injury model was established with acetaminophen.After a series of administration,the appearance of the liver was observed,then the liver and spleen were weighed and the liver index and the spleen index were evaluated;the levels of ALT and AST in the serum of the mice were determined using the kit;the pathological changes of the liver of the mice were observed by H＆E staining of paraffin sections;SOD and MDA levels in liver of mice were determined by kit;the contents of inflammatory factors such as TNF-α、IL-6 and NF-κB in mouse liver were determined using ELISA kits;the expression of HMGB1 and RAGE were analyzed by Western blot.After intraperitoneal injection of acetaminophen in mice,the liver had obvious congestion and edema;the levels of ALT and AST in the serum of the mice increased.In the group treated with LUT-DG,the serum ALT and AST levels returned to normal levels（compared with the healthy control group,P>0.05）,and there was no obvious edema and congestion in the liver and spleen;and the levels of inflammatory factors and oxidative stress in liver decreased.In addition,the expression of both HMGB1/RAGE proteins were significantly decreased（compared with PBS,P<0.05）.These results suggest that LUT-DG can significantly improve the oral bioavailability of LUT,and can effectively alleviate acetaminophen-induced liver damage by inhibiting oxidative stress and inflammatory cytokines.