| The phenomenon about drug-abuse has caused a series of problems with the development of modern medicine.A large number of drug-abuse to the hepatic metabolism bring a serious threat,cause drug induced hepatic injury in the proportion of hepatic diseases.Acetaminophen induced acute hepatic injury model in mice is one of the classic drug induced hepatic injury. In the scientific research field, the commonly used model is used to study the mechanism of hepatic injury, the decrease of enzyme and the protective drug development of hepatic injury. In-depth study of APAP induced hepatic injury in pathogenesis and development of effective hepatoprotective agents is a bottleneck in the contemporary development of medicine and health.In modern medicine, the extraction of flavonoids from different plants has been widely used in the protection of hepatic injury caused by various reasons, and early ago folk inspection records of folium syringae in the treatment of hepatic.But so far, there is no study on flavonoids of folium syringae hepatoprotective. This study based on the prophase of APAP drug-induced hepatic injury,to continue to carry out the hepatoprotective effect of flavonoids of folium syringae. Previous studies have found that phase II drug metabolism enzyme GSTA1 can be used as early diagnostic indicators of hepatic injury model, Therefore, this study also use GSTA1 as indicators of hepatic protective effect of flavonoids in the folium syringae, to lay the foundation for further study on the pathogenic mechanism of hepatic injury and hepatic protective drugs development.According to our previous research, we selected the leaves of folium syringae as the target of our study. In order to determine the effective protective components of folium syringae deeply,folium syringae reflux extraction with different concentrations of ethanol.The serum transaminase(ALT, AST) activity changes to determine the initial dissolution characteristics of effective components of protecting folium syringae.The ultrasonic assisted extraction method, ethanol reflux design of single factor experiment, observed in the case of single factor of different level of folium syringae flavonoids extraction effect. Then orthogonal design method was used to design the orthogonal design of three factors and four levels. The effect of various factors on the extraction of flavonoids from the folium syringae was studied by using the method of flavonoids as the index. order to determine the optimal extraction scheme of folium syringae flavonoids, the APAP induced acute hepatic injury model in the 40 mice were randomly divided into control group,model group, folium syringae flavonoids high dose group(40mg·kg-1), folium syringae flavonoids middle dose group(35mg·kg-1), folium syringae flavonoids low dose group(30mg·kg-1). Theserum transaminase(ALT, AST), hepatic index(GSH, GSH-Px, MDA, NO, SOD) phase II drug metabolizing enzyme GSTA1 and the pathological changes of the.To evaluate the protective effect of folium syringae flavonoids on acute hepatic injury in mice, and we observed the change trend in the whole process of injury and anti-injury in GSTA1 and sensitivity.Research results show that:(1)Single factor test and orthogonal design test of folium syringae flavonoids showed that the influence of each factor on the extraction of flavonoids was the size of the effect:liquid material ratio > extraction time > Ethanol concentration > extraction times. Combined with the optimal level of each factor and the actual production situation and orthogonal test results to determine the optimal extraction scheme: When the the ratio of liquid to material was 20:1,the ethanol concentration was 60%, the extraction time was 80 min, The number of extraction was 1 time,, the extraction rate was 127.46mg·g-1.(2) Protective test of folium syringae flavonoids on APAP induced acute hepatic injury in mice:Compared with model group, the level of serum aminotransferase(ALT, AST) activity was significantly decreased(p<0.01) in the folium syringae flavonoids dose group(40mg·kg-1). hepatic tissue index(MDA, NO) content was significantly decreased(p<0.01) in mice hepatic tissue index(GSH, GSH-Px, SOD) activity was significantly increased(p<0.01). Serum transaminase(ALT,AST) and hepatic index(GSH, GSH-Px, SOD, MDA, NO) was significantly(p< 0.05) or significant change(p<0.01) in folium syringae middle dose group and of low dose change in flavonoids of folium syringae group was less obvious.(3) Changes of phase II drug metabolicenzyme GSTA1 in APAP induced acute hepatic injury model in mice induced by folium syringae flavonoids.The content of GSTA1 in model group was significantly decreased(p<0.01) as Compared with the control group.The GSTA1 content in folium syringae flavonoids high dose group was significantly increased(p<0.01) as compared with model group.The content of GSTA1 in folium syringae flavonoids middle dose group was significantly higher(p<0.05), folium syringae flavonoids low dose group GSTA1 content change is not obvious.These changes were consistent with the changes of GSH, GSH-Px and SOD activity in liver tissues.(4)Hepatic tissue pathological results showed that the model group compared with the control group, the fatty degeneration of hepatic cells.In particular, the fat degeneration of the vesicles was obvious, and the cell nucleus lysis, disappearance, eosinophilic degeneration and inflammatory cell infiltration were obvious.Folium syringae folium group showed fatty degeneration dose significantly reduced or even disappeared, restore the cell structural integrity, morphology,appearance of binucleate cells.The effective repair of hepatic tissue damage. There was no significant difference in the repair of hepatic injury in the folium middle folium syringae dose group, but there was a significant difference between the model group and the model group.In this experiment, the model of acute hepatic injury induced by APAP in mice was successfully reproduced.And noted that clove alcohol extract is better than that of thehepatoprotective effect of aqueous extract, through single factor test and orthogonal test to optimize the extraction of flavonoids from the folium syringae, folium syringae flavonoids has a good protective effect on APAP induced acute hepatic injury in mice. GSTA1 showed regular changes in mice with acute hepatic injury,, Combined with the levels of ALT and AST in serum,hepatic index and hepatic tissue pathology, and determine that the folium syringae has high protective dose of 40mg·kg-1. |
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