Preparation Of Nano-fisetin And Its Protective Effect On Acute Liver Injury In Mice

ObjectiveDrug-induced liver injury is one of the most common acute liver injuries in clinical practice,and the most common cause of drug-induced liver injury is the overdose of acetaminophen.At present,N-acetylcysteine is the only clinical treatment for acute liver injury caused by acetaminophen,but N-acetylcysteine has a weak effect on advanced patients.Therefore,it has great significance to develop new therapeutic drug.In this study,a novel dipotassium glycyrrhizinate-fisetin premicelles（DG-FIT）was fabricated,acetaminophen was used to establish a mouse model of acute liver injury,and this animal model was used to evaluate the therapeutic effect of DG-FIT.Methods1.Solvent evaporation method was used to prepare DG-FIT,and the formulation of DG-FIT was optimized,and characterize its solids and solutions;the storage stability was evaluated;the antioxidant activity of DG-FIT was determined by FRAP method;the safety of DG-FIT was evaluated by hemolysis test.2.The solubility of DG-FIT in different media was determined,and the in vitro release characteristics were investigated by dialysis bag experiment;the permeability of DG-FIT was investigated by in vitro parallel artificial membrane permeation experiments;as a model drug,coumarin-6 was used to investigate its in vivo absorption characteristics by fluorescence intensity.3.A mouse model of liver injury was established using acetaminophen.The survival of the mice within 72 hours was observed;the liver and spleen of the mice were collected and weighed,the appearance of the liver was observed,the liver index and the spleen index were calculated;the serum ALT and AST of the mice were determined;the H＆E staining of paraffin sections was used to investigate the pathological changes of the liver;TUNEL method was used to evaluate the apoptosis of liver cells in mice;the levels of oxidative stress and inflammatory factors in the liver of mice were investigated.Results1.DG-FIT was successfully prepared.DG-FIT could be easily dissolved in water to form a clear and yellow micelle solution;the particle size of DG-FIT was84.17±3.88 nm,and the encapsulation efficiency reached 99.65±0.78 %;it had good storage stability and strong antioxidant capacity;this premicelles showed well safety with no hemolysis occurred.2.DG-FIT significantly increased the solubility of fisetin in water.In vitro release experiments,the accumulative release rate of premicelles reached92.87±2.79 % at 5 h;in vitro absorption experiments,the permeation amount of fisetin in the premicelles reached 267.30±14.53 μg at 3.5 h,which was 21.2 times that of the suspension group;in vivo absorption experiment showed that dipotassium glycyrrhizinate encapsulation significantly improved the intestinal absorption of model drug.3.A mouse liver injury model was successfully established by acetaminophen.The 72-hour survival rate of the DG-FIT group was 83.3 %,the appearance of the liver was similar to that of the healthy control group;the serum ALT and AST returned to normal levels（P>0.05 compared with the healthy control group）;compared with the control group,the levels of oxidative stress and inflammatory factors in the liver were significantly decreased（P<0.05）.ConclusionDG-FIT can significantly improved the solubility and in vivo absorption of the drug,and has a good development prospect for the protective effect of acetaminophen-induced acute liver injury in mice.