Studies On The Protective Effects And Mechanism Of Abrus Cantoniensis Hance Against APAP-Induced Liver Injury In Mice

Objective:ICR mice were intraperitoneally injected with APAP to establish a drug-induced acute liver injury model.The protective effect of the extract of Abrus cantoniensis Hance(AC)against APAP-induced liver injury was evaluated and its potential pharmacological mechanism was revealed.In addition,UPLC-Q/TOF-MS/MS was used to systematically characterize and identify the main components in the extract of AC.This study provided theoretical basis for the traditional use of AC in “eliminating dampness and removing yellow,clearing heat and detoxification”,and also provided a potential candidate drug for the prevention and treatment of APAP poisoning in clinic.Methods:The mice were pre-administrated for 7 days by intragastric administration of AC extract(100 or 200 mg/kg).Then,APAP(250 mg/kg)was injected intraperitoneally and treated for 12 h.Serum and liver tissues were collected to detect the activity of AST and ALT in serum and the levels of GSH,SOD,MDA,TNF-α,IL-1β and IL-10 in liver tissues.H&E and TUNEL staining were used to evaluate the degree of liver injury and apoptosis.The contents of i NOS,CYP2E1,SULT1A1,UGT1A1,Bcl-2,Bax,actin,Lamin B1,IκBα,p-IκBα,NF-κB P65,P38 MAPK,p-P38,JNK and p-JNK in liver homogenate were detected by western blot to elucidate its mechanism of action.At the same time,CCK-8 assay was used to detect the effects of the AC extracts on APAP-induced HepG2 cell viability in vitro.ROS levels and apoptosis in HepG2 cells were detected by flow cytometry.Finally,UPLC-Q/TOF-MS/MS was used to systematically characterize and identify the main components in the extract of AC.Results:After the administration of AC,it could significantly inhibit the APAP-induced oxidative stress and inflammation,and reduced the AST and ALT levels,thus improving the pathological injury of liver tissue and apoptosis,and presenting a significant protective effect on drug-induced liver injury induced by APAP.The cell experiment also confirmed that the extract could effectively inhibit the ROS production and hepatocyte apoptosis induced by APAP.Molecular mechanism studies showed that the administration of AC could significantly inhibit the activation of MAPK and NF-κB signaling pathways in mice liver.At the same time,AC extract could reduce the formation of active intermediate NAPQI by inhibiting the expression of CYP2E1 enzyme,and promote the expression of phase II metabolic enzymes SULT1A1 and UGT1A1 to make more APAP excreted in urine as non-toxic compounds.Finally,UPLC-Q/TOF-MS/MS method was used to systematically characterize and identify the main components in the extracts.Eight components,including trigonelline,Abrine,Vicenin-2,Vicenin Ⅲ,schaftoside,Isoschaftoside,Abrusamide B and Abrusamide A were preliminatively identified.Conclusion:This study first demonstrated that the extract of AC had an observable hepatoprotective activity on drug-induced liver injury induced by APAP,and its mechanism might be involved with inhibiting the activation of MAPK and NF-κB pathways,thus improving the liver injury caused by oxidative stress and inflammation.In addition,it could regulate the expression of APAP metabolism-related enzymes,and promoted the excretion and detoxification of APAP.The chemical profiling of AC extract was established by UPLC-Q/TOF-MS/MS,and the main components were identified,including trigonelline,Abrine,Vicenin-2,Vicenin Ⅲ,schaftoside,Isoschaftoside,Abrusamide B and Abrusamide A.The above results provided a theoretical basis for the traditional use of AC in “eliminating dampness and removing yellow,clearing heat and detoxification”,and also provided a new idea/potential candidate drug for APAP poisoning’s prevention and treatment in clinic.