Protective Effects And Mechanisms Of Isorhamnetin On Acetaminophen Induced Drug-induced Liver Injury In Mice

Objective:Drug-Induced Liver Injury(DILI)is caused by the drug itself or its metabolites during the process of drug usage.Drug induced strong toxicity is the main cause of acute liver failure.Isorhamnetin is a flavonoid substance extracted from natural plants.Researches showed that isorhamnetin possess good antioxidant,anti-inflammatory,and liver-protective effects.However,few research has been done on the effects of isorhamnetin on acetaminophen(APAP)-induced liver injury caused.In this experiment,APAP was used to induce liver injury in mice.The protective effects of isorhamnetin against APAP-induced liver injury and mechanisms was explored.The study may provide scientific basis for isorhamnetin to be used as a green liver-protecting drug.Methods:Male BALB/c mice were randomly divided into four groups(n=8):Control group(Control),APAP-induced liver injury group(APAP),isorhamnetin low-dose group(APAP+ISO30),isorhamnetin high-dose group(APAP+ISO90).Isorhamnetin(30,90mg/kg)was intragastrically administered to mice with 1%sodium carboxymethyl cellulose(CMC-Na)as a vehicle for 14 days.The Control group and the APAP group were given the same volume of 1%CMC-Na.After 14 days administration with isorhamnetin,APAP(300mg/kg)was given by intraperitoneal injection to induce liver injury.Liver and blood samples were collected 4 hours after APAP challenge.The levels of AST and ALT in serum were detected,and the liver sections were stained with HE to observe the pathological changes in the tissues.The changes of GSH,GSH-PX,MDA,SOD-2 in liver tissues.Real-time PCR was used to detect the expression of cytokines in the liver were measured.Western Blot was used to detect the expression of TLR4,p-ERK,p-JNK,and P62 protein.The expression of Keap-1,Nrf-2 and its downstream antioxidant factors HO-1,NQO-1,GCLC,SOD-2 were detected.Results:Isorhamnetin significantly reduced the content of AST and ALT in the serum,increasing the levels of GSH and GSH-PX in the liver,accompanied by decreased levels of MDA.Pathological examination showed that isorhamnetin significantly reduces the numbers of necrotic cells in the liver and effectively improve the liver damage caused by APAP.Western Blot and Real-time PCR tests showed that isorhamnetin effectively inhibits the expression of TLR4,p-ERK,and p-JNK,and reduces the expression of TNF-α,IL-1β,and IL-6 mRNA.Further results show that isorhamnetin can activate the Nrf-2 signaling pathway and promote the activation of antioxidant factors such as NQO-1.Conclusion:This experimental study shows that isorhamnetin can effectively alleviate APAP-induced drug-induced liver injury,and the protective effect is related to its anti-inflammatory and antioxidant functions.We speculate that isorhamnetin exerts anti-inflammatory and antioxidant effects against APAP-induced drug-induced liver damage by regulating MAPK/Nrf-2 signaling pathways.