定心方Ⅲ号方通过抑制Akt/mTOR/SREBP-1C通路改善ApoE^-/-小鼠非酒精性脂肪肝的实验研究  

Experimental Study of Dingxin Recipe Ⅲ Improving Non-alcoholic Fatty Liver in ApoE^-/-Mice by Inhibiting Akt/mTOR/SREBP-1C Signaling Pathway

作　　者：刘晓瑜[1] 徐煜凌 程赛博[1] 何培坤 顾民华 洪睦铿 周凤华[1] 贾钰华[1] Liu Xiaoyu;Xu Yuling;Cheng Saibo;He Peikun;Gu Minhua;Hong Mukeng;Zhou Fenghua;Jia Yuhua(College of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515)

机构地区：[1]南方医科大学中医药学院,广州510515

出　　处：《中药药理与临床》2020年第5期149-154,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：广州市科技计划产学研协同创新重大专项项目(编号:201704020042);国家自然科学基金(编号:81973802、81373574);广东省名优中成药二次开发项目(编号:20174010);广东省省级科技计划项目(编号:2016A020226002)。

摘　　要：目的:观察定心方Ⅲ号方对高脂饮食诱导的ApoE^-/-小鼠非酒精性脂肪肝(NAFLD)的影响。方法:40只雄性ApoE^-/-小鼠随机分为模型对照组、定心方Ⅲ号方6 g/kg、12 g/kg、24 g/kg组和辛伐他汀5 mg/kg组,每组8只,并设同龄C57BL/6J雄性小鼠为正常对照组。ApoE^-/-小鼠连续喂养高脂饲料12 w复制NAFLD模型后,各组分别连续给药12 w。末次给药后禁食8 h,检测小鼠空腹血糖、体质量和肝湿质量,计算肝指数;采用生化法检测小鼠血清总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C),酶标法检测小鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、还原型谷胱甘肽(GSH)、丙二醛(MDA)浓度和超氧化物歧化酶(SOD)活力,HE染色观察小鼠肝脏病理形态,Western Blot检测小鼠肝脏中磷酸化蛋白激酶B(p-Akt)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)、脂肪酸合成基因(SREBP-1C)、核孔糖蛋白62(p62)、微管相关蛋白轻链3Ⅱ(LC3Ⅱ)蛋白的表达情况。结果:与正常对照组小鼠比较,模型对照组小鼠的血糖、体质量、肝湿质量和肝指数明显升高(P<0.05或P<0.01),肝细胞中脂滴明显聚集,肝索排列紊乱或消失,NAFLD病理评分显著升高(P<0.01),血清中TC、TG、LDL-C、HDL-C、ALT、AST和MDA的浓度明显升高(P<0.05或P<0.01),SOD和GSH活性显著降低(P<0.01),肝组织中p-Akt、p-mTOR、SREBP-1C、p62、LC3Ⅱ蛋白表达均明显升高(P<0.05或P<0.01);与模型对照组小鼠比较,给药组小鼠血糖、体质量、肝湿质量和肝指数均降低,肝组织病理形态学明显改善,NAFLD评分显著下降,血清中TC、LDL-C、TG、ALT、AST和MDA浓度均有不同程度的下降,HDL-C、SOD和GSH含量活性均升高,肝组织中p-Akt、p-mTOR、SREBP-1C、p62、LC3Ⅱ蛋白表达均明显下降,与模型对照组比较,其中以定心方Ⅲ号方24 g/kg组和辛伐辛伐他汀5 mg/kg组变化最为明显(P<0.05或P<0.01)。结论:定心方Ⅲ号方可降�Objective:To observe the effect of Dingxin Recipe Ⅲ on non alcoholic fatty liver disease(NAFLD)induced by high-fat diet in ApoE^-/-mice.Methods:Forty male ApoE^-/-mice were randomly divided into the model group,different doses of Dingxin Recipe Ⅲ groups(6 g/kg,12 g/kg,24 g/kg)and Simvastatin group(5 mg/kg),8 rats in each group,and the same age C57BL/6J male mice were used as the control group.ApoE^-/-mice were continuously fed with high-fat diet for 12 weeks to establish a non-alcoholic fatty liver model,then all rats were orally administered with corresponding drugs for 12 weeks.8 hours after the last administration,the fasting blood glucose,the body weight and the wet weight of the liver were measured to calculate the liver index.The serum levels of TC,TG,HDL-C and LDL-C were detected by the biochemical method,the levels of ALT,AST,GSH,MDA and SOD in serum were detected by enzyme labeling.HE staining was used to observe the pathological lesions of the liver.Western blot were used to detect the expressions of p-Akt、p-mTOR、SREBP-1C、p62、LC3Ⅱprotein in the liver.Results:Compared with the control group,the blood glucose,the body weight,the wet weight of liver and the liver index in the model group were significantly increased(P<0.01 or P<0.05).The lipid droplets in liver cells were significantly accumulated,the arrangement of hepatic cords was disordered or disappeared,the pathological score of NAFLD was significantly increased(P<0.01).The contents of TC,TG,LDL-C,HDL-C,ALT,AST and MDA in serum were increased(P<0.01 or P<0.05),the activities of SOD and GSH were decreased(P<0.01),and the expressions of p-Akt,p-mTOR,SREBP-1c,p62 and LC3Ⅱproteins in liver tissues were all increased(P<0.01 or P<0.05).Compared with the model group,the blood glucose,the body mass,the wet mass of liver and the liver index of the mice in the administration group were reduced,the pathological morphology of liver tissues was significantly improved,the NAFLD score was significantly decreased,and the serum contents of TC,LDL-C,TG

关 键 词：定心方Ⅲ号方 非酒精性脂肪肝 氧化应激 自噬 蛋白激酶/雷帕霉素靶蛋白/脂肪酸合成基因信号通路 

分 类 号：R285.5[医药卫生—中药学]