黄芪葛根汤与瓜蒌汤合方对糖尿病动脉粥样硬化大鼠主动脉SIRT1/AMPK信号通路的影响

Effects of Huangqi Gegen Decoction combined with Gualou Decoction on SIRT1/AMPK signaling pathway in aorta of diabetic atherosclerosis rats

目的:基于沉默信息调节因子1/AMP活化蛋白激酶(SIRT1/AMPK)信号通路探讨黄芪葛根汤与瓜蒌汤合方降低糖尿病动脉粥样硬化大鼠血糖、血脂、氧化应激的作用机制。方法:将68只SPF级雄性SD大鼠按随机血糖分为空白组、模型组、TMK组(糖脉康颗粒)、汤剂组(黄芪葛根汤与瓜蒌汤合方汤剂)、颗粒剂组(黄芪葛根汤与瓜蒌汤合方颗粒剂)。除空白组外,其余各组喂养28 d高脂饲料,第29天按30 mg/kg一次性腹腔注射2%链脲佐菌素(STZ)诱导糖尿病动脉粥样硬化大鼠模型,注射STZ第3天灌胃给予相应药物或蒸馏水,连续灌胃49 d。检测血糖、甘油三酯(TG)、胆固醇(CHO)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C),ELISA检测丙二醇(MDA)、活性氧(ROS)、超氧化物歧化酶(SOD)、糖化血红蛋白(HbA1c)含量,Real-time PCR法检测AdipoR1、SIRT1、AMPK、GLUT4、SREBP-1c mRNA相对表达量。结果:与空白组比较,模型组治疗后血糖、TG、CHO、LDL-C含量显著升高(P<0.05),HDL-C含量显著降低(P<0.05),MDA、ROS、HbA1c含量显著升高(P<0.05),SOD含量显著降低(P<0.05),SIRT1、AMPK、GLUT4、AdipoR1 mRNA表达量显著降低(P<0.05),SREBP-1c mRNA表达量显著增加(P<0.05)。与模型组比较,各给药组能降低大鼠血糖、TG、CHO、LDL-C、MDA、ROS、HbA1c含量(P<0.05),升高HDL-C、SOD含量(P<0.05),提升SIRT1、AMPK、GLUT4、AdipoR1 mRNA表达量(P<0.05),降低SREBP-1c mRNA表达量(P<0.05)。结论:黄芪葛根汤与瓜蒌汤合方降低糖尿病动脉粥样硬化大鼠血糖血脂可能与刺激AdipoR1分泌,激活SIRT1/AMPK信号通路,调节下游因子GLUT4、SREBP-1c的活性有关。黄芪葛根汤与瓜蒌汤合方降低氧化应激反应可能与降低ROS、MDA表达,增加SOD、SIRT1表达有关。

Objective:To explore the mechanism of Huangqi Gegen Decoction combined with Gualou Decoction in reducing blood glucose,blood lipid and oxidative stress in diabetes atherosclerotic rats based on the signal pathway of silent information regulator 1/AMP-activated protein kinase(SIRT1/AMPK).Methods:A total of 68 SPF male SD rats were randomly divided into blank group,model group,TMK group(Tangmaikang Granules),decoction group(Huangqi Gegen Decoction combined with Gualou Decoction),granule group(Huangqi Gegen Decoction and Gualou Decoction combined granule).Except for the blank group,each group was fed with fat diet for 28 days.On the 29th day,the rat model of diabetes atherosclerosis was induced by intraperitoneal injection of 2%STZ at the dose of 30 mg/kg.The corresponding drug or distilled water were given by gavage on the third day of injection of STZ,and the rats were gavaged continuously for 49 days.Biochemical analyzer detected blood glucose,triglyceride(TG),cholesterol(CHO),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),and ELISA detected the contents of propylene glycol(MDA),reactive oxygen species(ROS),superoxide dismutase(SOD)and glycosylated hemoglobin(HbA1c).Adiponectin receptor(AdipoR1),silent information regulator 1(SIRT1),adenylate activated protein kinase(AMPK),glucose transporter 4(GLUT4)and sterol regulatory element binding protein-1c(SREBP-1c)mRNA were detected by Real-time PCR.Results:Compared with the blank group,the contents of Glu,TG,CHO and LDL-C in the model group increased significantly(P<0.05),the contents of HDL-C decreased significantly(P<0.05),the contents of MDA,ROS and HbA1c increased(P<0.05),the content of SOD decreased(P<0.05),the expression of SIRT1,AMPK,GLUT4 and AdipoR1 mRNA decreased(P<0.05),and the expression of SREBP-1c mRNA increased(P<0.05).Compared with the model group,the giving drug groups could reduce the contents of Glu,TG,CHO,LDL-C,MDA,ROS and HbA1c in diabetes atherosclerotic rats(P<0.05),increase the contents of HDL-C and SOD(P<0.05),increase the mRNA of SIRT1,AMPK,GLUT4 and AdipoR1(P<0.05),and reduce the expression of SREBP-1c mRNA(P<0.05).Conclusion:Huangqi Gegen Decoction combined with Gualou Decoction reduce blood glucose and lipids in diabetes atherosclerotic rats,which may be related to stimulating the secretion of AdipoR1,activating SIRT1/AMPK signal pathway,and regulating the activities of downstream factors GLUT4 and SREBP-1c.Huangqi Gegen Decoction combined with Gualou Decoction may reduce the oxidative stress response by reducing ROS,MDA and increasing the expression of SOD and SIRT1.