橙皮苷对果糖诱导的非酒精性脂肪肝病小鼠肝脏组织中CYP2E1表达的影响

Effect of hesperidin on CYP2E1 expression in nonalcoholic fatty liver rats induced by high fructose diet

目的探讨橙皮苷对果糖+高脂饮食构建的非酒精性脂肪肝病(NAFLD)小鼠的干预作用及作用机制。方法60只雄性昆明小鼠适应性喂养1周后按照随机数字表法分为正常组、模型组、多烯磷脂酰胆碱组、橙皮苷低剂量组、橙皮苷中剂量组和橙皮苷高剂量组各10只。除正常组外,其余组小鼠均采用高脂饲料+10%果糖水日常饮用8周构建NAFLD模型,然后多烯磷脂酰胆碱组给予多烯磷脂酰胆碱23 mg/(kg·d)灌胃,橙皮苷低、中、高剂量组分别给予80 mg/(kg·d)、160 mg/(kg·d)和320 mg/(kg·d)灌胃橙皮苷灌胃。灌胃4周后采血检测各组小鼠肝功能指标[谷草转氨酶(AST)、谷丙转氨酶(ALT)]、血脂指标[高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)]、氧化应激指标[谷胱甘肽(GSH)、超氧化物歧化酶(SOD)],观察各组小鼠肝脏病理学变化,实时荧光定量PCR和Western blot法检测肝脏组织中细胞色素P4502E1(CYP2E1)mRNA和蛋白表达情况。结果与正常组比较,模型组小鼠血清AST、ALT、HDL-C和LDL-C水平均明显升高(P均<0.05),GSH和SOD水平均明显降低(P均<0.05);与模型组比较,各药物组小鼠血清AST、ALT和LDL-C水平均明显降低(P均<0.05),HDL-C水平均无明显变化(P均>0.05),GSH和SOD水平均明显升高(P均<0.05)。模型组小鼠肝脏组织有明显脂肪空泡,而各药物组脂肪空泡数量较少。与正常组比较,模型组小鼠肝脏组织中CYP2E1蛋白及mRNA表达水平均明显升高(P均<0.05);与模型组比较,各药物组小鼠肝脏组织中CYP2E1蛋白及mRNA表达水平均明显降低(P均<0.05),但橙皮苷各剂量组仍明显高于多烯磷脂酰胆碱组(P均<0.05)。结论橙皮苷对NAFLD小鼠有明显治疗作用,其综合疗效与多烯磷脂酰胆碱相似,CYP2E1可能是其作用的一个靶点。

Objective It is to observe the intervention effect of hesperidin on the non-alcoholic fatty liver disease(NAFLD)mice constructed by fructose+high-fat diet and research its mechanism.Methods After one week of adaptive feeding,60 Kunming mice were divided into 6 groups via random number method,which were the normal group,model group,polyene phosphatidylcholine group,and low-dose,medium-dose and high-dose group of nobiletin,10 mice in each group.Except for the normal group,the mice in the other groups were fed with high-fat diet+10%fructose water daily for 8 weeks to build NAFLD models,and then the polyene phosphatidylcholine group was given polyene phosphatidylcholine 23 mg/(kg·d)by gavage,low-dose,medium-dose and high-dose groups of nobiletin were given 80 mg/(kg·d),160 mg/(kg·d)and 320 mg/(kg·d)hesperidin by gavage.After 4 weeks of intragastric administration,the blood of the mice in each group was collected to detect liver function indexes[aspartate aminotransferase(AST),alanine aminotransferase(ALT)],blood lipid indexes[high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)],oxidative stress indicators[glutathione(GSH),superoxide dismutase(SOD)],the pathological changes in the liver of the mice in each group were observed,and Real-time fluorescent quantitative PCR and Western blot were used to detect the expression of cytochrome P4502E1(CYP2E1)mRNA and protein in liver tissues.Result Compared with the normal group,the levels of serum AST,ALT,HDL-C,LDL-C of mice in the model group were significantly increased(P<0.05),GSH and SOD were significantly decreased(P<0.05);compared with the model group,the levels of serum AST,ALT,LDL-C of the mice were significantly lower in each treatment group(P<0.05),there was no significant difference in HDL-C,and the levels of GSH and SOD were significantly higher in each treatment group(P<0.05).The liver tissues of mice in the model group had obvious fatty vacuoles,while the number of fatty vacuoles in each treatment group was less.Compared with the normal group,the expression of CYP2E1 protein and mRNA in the liver tissue of the model group was significantly increased(P<0.05);compared with the model group,the expression of CYP2E1 protein and mRNA in the liver tissue of the mice in each treatment group was significantly decreased(all P<0.05),but each treatment group of hesperidin was still significantly higher than the polyene phosphatidylcholine group(all P<0.05).Conclusion Hesperidin has a significant therapeutic effect on NAFLD mice,and it have the similar effect with the polyene phosphatidylcholine,CYP2E1 may be a target of Hesperidin.