当飞利肝宁胶囊调控肝脏氧化应激改善非酒精性脂肪性肝病小鼠的机制研究  

Mechanism of Dangfei Liganning Capsule on non-alcoholic fatty liver disease via inhibiting liver oxidative stress in mice

作　　者：赵燕婷 舒祥兵 杨志新[1] 李光萍[1] ZHAO Yanting;SHU Xiangbing;YANG Zhixin;LI Guangping(Baoshan Branch of Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai Baoshan Integrated Traditional Chinese and Western Medicine Hospital,Shanghai 201999,China)

机构地区：[1]上海中医药大学附属曙光医院宝山分院,上海市宝山区中西医结合医院,上海201999

出　　处：《上海中医药大学学报》2021年第5期37-43,74,共8页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81804020)。

摘　　要：目的:探讨当飞利肝宁胶囊对高脂饮食诱导的非酒精性脂肪性肝病(NAFLD)小鼠的治疗作用及潜在机制。方法:雄性C57BL/6J小鼠随机分为正常组、模型组和当飞利肝宁(0.42 g·kg^(-1)·d^(-1))组,每组10只。除正常组外,其他小鼠给予高脂饲料喂养12周,以诱导模型。造模后,各组小鼠灌胃给予相应药物干预,连续4周。末次给药后,取血清,收集肝组织。生化分析仪检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)水平;油红O染色和HE染色后观察肝组织形态学变化。试剂盒检测肝组织TC、TG、丙二醛(MDA)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)水平;PCR检测肝脏核因子E2相关因子2(Nrf2)、细胞色素P450家族成员2E1(CYP2E1)、血红素氧合酶1(HO-1)、NADPH醌氧化还原酶(NQO1)、谷胱甘肽硫转移酶P1(GSTP1)的m RNA表达;Western blot检测肝脏Nrf2和CYP2E1的蛋白表达。结果:①与模型组相比,当飞利肝宁组小鼠血清ALT、AST、TC和LDL水平显著降低(P<0.05)。②当飞利肝宁胶囊能明显降低模型小鼠的肝脏TG水平(P<0.05),且明显减轻模型小鼠肝组织肝细胞脂肪变性、脂质堆积;与模型组相比,当飞利肝宁组小鼠肝脏SOD活性明显升高(P<0.05),MDA含量显著降低(P<0.05)。③模型组小鼠肝脏Nrf2、HO-1、GSTP1、NQO1及CYP2E1的m RNA表达水平较正常组均显著下降(P<0.05,P<0.01),当飞利肝宁胶囊干预可明显上调模型小鼠肝脏Nrf2、GSTP1及NQO1的m RNA表达水平(P<0.05)。④模型组小鼠肝脏Nrf2和CYP2E1的蛋白表达水平较正常组明显降低(P<0.05,P<0.01),当飞利肝宁胶囊干预可明显升高模型小鼠肝脏Nrf2和CYP2E1的蛋白表达水平(P<0.05)。结论:当飞利肝宁胶囊对高脂饮食诱导的NAFLD小鼠具有良好的治疗作用,其作用机制与降低Nrf2介导的肝脏氧化应激水平有关。Objective:To investigate the therapeutic effects and potential mechanisms of Dangfei Liganning Capsule on high fat diet induced non-alcoholic fatty liver disease(NAFLD)in mice.Methods:Male C57BL/6J mice were randomly divided into the normal group,model group and Dangfei Liganning(0.42 g·kg^(-1)·d^(-1))group,10 mice in each group.Except the normal group,mice in the other groups were fed with high fat diet for 12 weeks to induce the model.After modeling,each group was treated with the corresponding drug by intragastric administration for 4 weeks.After the last administration,the serum was taken and the liver tissue was collected.The serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG),low-density lipoprotein(LDL)and high-density lipoprotein(HDL)were detected by biochemical analyzer.The morphological changes of liver tissue were observed after oil red O staining and HE staining.The levels of TC,TG,malondialdehyde(MDA),glutathione(GSH)and superoxide dismutase(SOD)in liver tissue were detected by kits.The m RNA expressions of nuclear factor erythroid 2-related factor 2(Nrf2),cytochrome P4502E1(CYP2E1),heme oxygenase 1(HO-1),NADPH quinone oxidoreductase 1(NQO1)and glutathione S-transferase P1(GSTP1)in liver tissue were detected by PCR.The protein expressions of Nrf2 and CYP2E1 in liver tissue were detected by Western blot.Results:①Compared with the model group,the serum levels of ALT,AST,TC and LDL were significantly decreased in the Dangfei Liganning group(P<0.05).②Dangfei Liganning Capsule could significantly reduce the TG level in the liver of model mice(P<0.05),and significantly alleviate the hepatocyte steatosis and lipid accumulation in the model mice.Compared with the model group,the activity of SOD in the liver was increased obviously in the Dangfei Liganning group(P<0.05),and the content of MDA in the liver was significantly decreased(P<0.05).③The m RNA expression levels of Nrf2,HO-1,GSTP1,NQO1 and CYP2E1 in the model group were significan

关 键 词：当飞利肝宁胶囊 非酒精性脂肪性肝病 氧化应激 核因子E2相关因子2 小鼠 

分 类 号：R285.5[医药卫生—中药学]