The Protective Effect And Mechanism Of Schizandrin Lignin On Drug-induced Liver Injury

Objective:Acetaminophen(APAP)is a widely used analgesic and antipyretic drug.Its overdose is the main cause of drug-induced liver injury.It is relatively safe at the recommended dose,but acute or cumulative overdose often leads to liver injury.Therefore,further elucidating the cellular and molecular mechanisms of liver injury caused by APAP can help to identify risk factors for acute liver failure and poor prognosis as soon as possible,so as to develop targeted therapeutic targets to block the process of liver injury;Drug-induced liver injury(DILI),also known as drug-induced liver disease,is the liver caused by toxic damage or allergic reactions caused by the drug or metabolites produced by the drug when the drug is applied at a therapeutic dose.disease.Schisandra is a traditional liver-protecting Chinese medicine.The literature shows that the alcohol extracts and polysaccharides of Schisandra have the effect of protecting the liver.Schisandra is widely used in the treatment of liver diseases,but there are few studies on the treatment of schisandra lignans in the treatment of APAP-induced DILI,and schisandra lignans the research mechanism of the hepatoprotective effects of the vegetarian ingredients is still unclear.Based on the results of the previous research of the research group,the alcohol extract of Schisandra can effectively treat the DILI caused by APAP in mice,but its medicinal material basis needs to be further studied.At present,many drugs in clinical application can cause drug-induced liver injury.Drug-induced liver injury is one of the main types of adverse drug reactions.Therefore,the prevention and treatment of liver damage caused by drugs in clinical use is particularly important.Methods:1.Purify and separate the total lignans of Schisandra by using the macroporous adsorption resin AB-8,detect the content of the total lignans by UV,use acetonitrile-water as the mobile phase,and use a gradient elution method at a wavelength of 217 nm.The total lignans of Schisandra chinensis before and after purification were detected by HPLC method.2.The DILI model of ICR mice was modeled at a dose of 200 mg/kg APAP,and the model dose of APAP was determined by measuring the AST and ALT indicators in the serum.3.Divide ICR mice into a blank group,a model group,a positive control group,and a low,medium and high total lignans of Schisandra chinensis.According to the grouping,different corresponding drug treatments are given to the mice for liver pre-protection.APAP was used to establish a mouse DILI model,the Lai’s detection method was used to determine the traditional biochemical indicators of AST and ALT in the mouse serum,the inflammation levels of TNF-α,IL-1β and IL-6,and the MDA and SOD in the mouse liver tissue homogenate.Oxidative stress and lipid peroxidation indicators.4.Combining the basis of previous research,taking 15 key genes screened out by PCR results of schisandra chinensis alcohol extract as the research objects,real-time PCR verification was carried out to verify the pathway and mechanism of schisandra chinensis total lignans in the treatment of APAP-induced DILI.5.Combining the results of the efficacy and PCR verification,Western blot verification of the key proteins in the ERBB signaling pathway was performed,respectively,for the blank group,model group,positive drug group,schisandra alcohol extract group and schisandra lignan group EGFR,LPCAT1 and the expression of MDM2 protein to verify the pathway and mechanism of schisandra in the treatment of APAP-induced DILI.6.Based on molecular docking technology,based on 12 related proteins,Schisandrol B(Sol B),Schisantherin A(Stn A),Schisandrin A(Sch A),Schisandrin phenol,Schisandrin C(Sch C),Schisandrol A(Sol A)and Schisandrin B(Sch B)Carry out ingredient screening to screen the important active ingredients of Schisandra lignans in the treatment of DILI.7.Make a model of ICR mice at a dose of 200 mg/kg APAP,copy the model,divide the ICR mice into blank group,model group,positive control group,Sch C group,Sol A group and Stn A group,and give different corresponding groups according to the group.Drug treatment,liver pre-protection by administration of mice,and use APAP to establish mouse DILI model,Lai’s detection method to determine the traditional biochemical indicators of AST and ALT in mouseserum,TNF-α,IL-1β and IL-6 inflammation Levels,and determine the MDA and SOD oxidative stress and lipid peroxidation indicators in the mouse liver tissue homogenate.Results:1.In this study,AB-8 type macroporous adsorption resin was used,the concentration of the sample liquid was 1.5mg/m L,the flow rate was 2BV/h,the diameter-to-height ratio was above 1:7,the elution flow rate was 2BV/h with 95% ethanol.Purification process purifies and separates Schisandra lignans.Based on the optimization of the purification process,a large number of schisandra lignans are enriched.Sol A is used as the key indicator for the purification of the total lignans of Schisandra.At the same time,HPLC method was used to determine seven kinds of lignans in Schisandra chinensis and compare the changes of lignans in the alcohol extract of Schisandra chinensis with those after purification.The results showed that AB-8 macroporous adsorption resin effectively purified the lignans in Schisandra chinensis,Laid the foundation for the enrichment of Schisandra lignans.2.Model with 200mg/kg APAP dose,pre-protection of Schisandra total lignans.Compared with the model group,Schizandrin total lignans have a significant therapeutic effect on APAPinduced DILI.Schisandra total lignans are pre-protected by administration of total lignans.The group can significantly reduce the level of AST,ALT,MDA,increase the level of SOD,and improve the DILI induced by APAP.It can treat the DILI caused by APAP by reducing the liver enzyme indicators in the serum,resisting oxidative stress and reducing inflammation.3.Based on the foundation of the previous research,a total of 15 targets including MDM2,SRC,PLA2 and EGFR were determined,and 15 target genes were finally determined for PCR verification.The results showed that the lignan treatment of APAP induced DILI signal in ERBB the key proteins EGFR,LPCAT1 and MDM2 in the ERBB signaling pathway were then selected for Western blot verification.The results showed that the total lignans of Schisandra can prevent and treat APAP-induced DILI by regulating the expression of EGFR and LPCAT1 proteins.4.Based on molecular docking technology,the 7 lignans were screened,and the results showed that the biphenylcyclooctene lignans in Schisandra: Sch C,Sol A and Stn A play a key role in the treatment of APAP-induced DILI;pharmacodynamic research It shows that Sch C,Sol A and Stn A can reduce the level of AST and ALT in the serum of DILI mice,increase the level of TNF-α,and regulate the expression of inflammatory factors in mice with liver injury.Conclusion:1.AB-8 type macroporous adsorption resin has the best effect on the separation and purification of lignans in the alcohol extract of Schisandra chinensis.After purification,the total lignans account for more than 60% of the solids.The purification process selected in this study has improved Schisandra chinensis the purity of lipotin.2.Schisandra lignans have obvious therapeutic effects on APAP-induced DILI,mainly by reducing liver enzymes in serum,resisting oxidative stress,lipid peroxidation and reducing inflammation to play the role of APAP-induced DILI.3.The total lignans of Schisandra are the effective components of Schisandra chinensis that play a role in protecting the liver and liver.It is mainly used to regulate the expression of EGFR and LPCAT1 related proteins in the ERBB signaling pathway to reduce the DILI caused by APAP.It is Schisandra lignin.The research on the treatment of APAP-induced DILI has laid the foundation.4.Based on molecular docking technology,the biphenylcyclooctene lignans in Schisandra have been screened out: Sch C,Sol A and Stn A play a key role in the treatment of APAP-induced DILI;Sch C and Sol A can reduce serum liver enzyme indicators and resist oxidation to stimulate and reduce inflammation to play a role in the treatment of APAP-induced DILI.