葛根素调控氧化应激与炎症反应改善小鼠脂肪性肝病OA

Objective To study the therapeutic effects of puerarin against metabolic-associated fatty liver disease(MAFLD)and the related underlying mechanism.Methods Forty-two C57BL/6J mice were randomly divided into six groups,i.e.,normal control group,model group,low-dose puerarin group(75 mg/kg),medium-dose puerarin group(150 mg/kg),and high-dose puerarin group(300 mg/kg),and metformin group(300 mg/kg)using random number table,7 in each group.To construct MAFLD models,mice were fed by high fat diet for 12 weeks,and mice received corresponding drugs by gastrogavage for 4 successive weeks.By the end of the intervention,the serum sample was collected from the mice eyeballs.Then mice were sacrificed by cervical dislocation,and livers were dissected,weighed to calculate the liver index.Serum concentrations of alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglycerides(TG),and total cholesterol(TC),as well as hepatic levels of malondialdehyde(MDA)and superoxide dismutase(SOD),were quantified using enzyme-linked immunosorbent assay(ELISA).Pathological changes of the liver tissue were examined by Hematoxylin-Eosin(HE)staining.mRNA levels of interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in the liver tissue were measured by RT-qPCR.Protein levels of AMP-activated protein kinase(AMPK),p-AMPK,nuclear factor erythroid 2 related factor 2(Nrf2),heme oxygenase-1(HO-1),nuclear factor Kappa B(NF-κB),p-NF-κ B were measured by Western Blot.Results Compared with the normal control group,mice in the model group had higher body/liver weight and liver index(P＜0.01).Significant hepatic steatosis and increased inflammatory cell infiltration occurred in the model group.Additionally,serum concentrations of TC,TG,ALT,and AST significantly increased(P＜0.01).MDA levels in the liver and mRNA levels of IL-6 and TNF-αincreased,while SOD levels decreased(P＜0.01).In the model group,p-AMPK,Nrf2,and HO-1 protein expressions significantly decreased,whereas p-NF-κB protein expression significantly increased(P＜0.05,P＜0.01).Compared with the model group,the body weight,liver weight,and liver index of mice in all intervention groups significantly decreased(P＜0.05,P＜0.01).Hepatic steatosis were improvd and inflammatory cell infiltration decreased,serum TC and TG levels decreased(P＜0.05,P＜0.01),HO-1 protein expression increased in all the intervention groups(P＜0.05,P＜0.01).Except for the low-dose puerarin group,decreased serum ALT levels and liver MDA levels,and increased SOD levels occurred in the other intervention groups(P＜0.05,P＜0.01).Decreased serum AST level occurred in medium-dose puerarin group and metformin group(P＜0.01).Decreased mRNA expression levels of IL-6 and TNF-αoccurred in liver(P＜0.05,P＜0.01).Increased liver protein levels of p-AMPK and Nrf2,and decreased p-NF-κB protein level occurred as well(P＜0.05,P＜0.01).The above-mentioned indices showed some improvement in the low-dose and high-dose puerarin groups,but with nostatistical differences(P＞0.05).Conclusion Puerarin significantly ameliorated liver lipid deposition,enhanced liver anti-oxidative capabilities,and attenuated hepatic inflammation in MAFLD model mice,which might be related to regulating AMPK/Nrf2/NF-κB signaling pathways.