地黄饮子“异病同治”糖尿病脑病和阿尔茨海默病脑胰岛素抵抗的机制研究  

Study on the mechanism of brain insulin resistance in diabetic encephalopathy and Alzheimer's disease by"homotherapy for heteropathy"with DihuangYinzi

作　　者：武东燕 王小丹 桂婉威 柴金苗 李钦青 贺文彬 张俊龙 WU Dong-yan;WANG Xiao-dan;GUI Wan-wei;CHAI Jin-miao;LI Qin-qing;HE Wen-bin;ZHANG Jun-long(Shanxi University of Chinese Medicine,Jinzhong Shanxi 030600,China)

机构地区：[1]山西中医药大学,山西晋中030600

出　　处：《时珍国医国药》2024年第8期1843-1848,共6页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(81874420,82274388);山西省中医药管理局课题(2022ZYYC099,2022ZYYC268)。

摘　　要：目的 观察地黄饮子对db/db小鼠和APP/PS1小鼠的治疗作用,探讨地黄饮子“异病同治”改善糖尿病脑病(Diabetic encephalopathy, DE)和阿尔茨海默病(Alzheimer’s disease, AD)脑胰岛素抵抗的分子机制。方法 分别设置DE疾病模型组和AD疾病模型组。DE疾病模型组中,10只db/m小鼠作为空白对照组(db/m组),30只db/db小鼠随机分为模型组(db/db组)、地黄饮子组(db/db+DHYZ组,30.03 g/kg)、二甲双胍组(db/db+MET组,0.61 g/kg)。AD疾病模型组中,10只野生型C57BL/6J小鼠作为空白对照组(C57组),30只APP/PS1小鼠随机分为模型组(APP/PS1组)、地黄饮子治疗组(APP/PS1+DHYZ组,30.03 g/kg)、美金刚治疗组(APP/PS1+MEM组,6.1 mg/kg)。药物干预4周后,葡萄糖氧化酶法和酶联免疫吸附法(ELISA)分别检测小鼠海马葡萄糖和胰岛素含量;苏木素-伊红(HE)染色观察小鼠海马组织病理改变;蛋白免疫印迹(Western blot)法检测海马磷脂酰肌醇-3-羟激酶(phosphoinositide 3-kinase, PI3K)、蛋白激酶B(Protein kinase B, Akt1)、磷酸化蛋白激酶B(Phospho-protein kinase B, pAkt1)蛋白表达量;免疫组化法检测海马组织胰岛素受体底物1(Insulin receptor substrate 1, IRS1)、葡萄糖转运蛋白4(Glucose transporter 4, GLUT4)表达。结果 Db/db小鼠和APP/PS1小鼠海马均出现神经元排列紊乱,细胞数目显著减少,核仁结构不完整等病理改变;葡萄糖含量均升高(P<0.05);db/db小鼠海马胰岛素升高(P<0.01)而APP/PS1小鼠海马胰岛素降低(P<0.01);PI3K、pAkt1/Akt1、GLUT4蛋白表达量均降低(P<0.01),IRS1蛋白表达量显著升高(P<0.01)。地黄饮子干预后db/db小鼠和APP/PS1小鼠的海马病理改变减轻,细胞数量增加;葡萄糖含量均降低(P<0.05);db/db小鼠海马胰岛素降低(P<0.01),而APP/PS1小鼠海马胰岛素升高(P<0.05);PI3K、pAkt/Akt、GLUT4蛋白表达量升高(P<0.05,P<0.01),IRS1蛋白表达量降低(P<0.05,P<0.01)。结论 地黄饮子可通过调节胰岛素相关IRS1/PI3K/Akt/GLUT4信号通路Objective We observed the therapeutic effects of Dihuang Yinzi on db/db mice and APP/PSl mice,and investigated the molecular mechanism of Dihuang Yinzi to improve the brain insulin resistance in diabetic encephalopathy(DE)and Alzheimer's disease(AD)by the"homotherapy for heteropathy".Methods In this study,we set up the DE disease model group and the AD disease model group,respectively.In the DE disease model group,10 db/m mice were used as a blank control group(db/m group),and 30 db/db mice were randomly divided into the model group(db/db group),the Dihuang Yinzi group(db/db+DHYZ group,30.03 g/kg),the metformin group(db/db+MET group,0.61 g/kg).In the AD disease model group,10 wild-type C57BL/6J mice were used as a blank control group(C57 group),and 30 APP/PS1 mice were randomly divided into a model group(APP/PS1 group),a Dihuang Yinzi treatment group(APP/PS1+DHYZ group,30.03 g/kg),and a memantine treatment group(APP/PS1+MEM group,6.1 mg/kg).After 4 weeks of drug intervention,mice hippocampus glucose and in-sulin levels were measured by glucose oxidase assay and enzyme-linked immunosorbent assay(ELISA),respectively;his-topathological changes in mice hippocampus were observed by hematoxylin-eosin(HE)staining;and hippocampus phos-phoinositide 3-kinase(PI3K),protein kinase B(Aktl),and phospho-protein kinase B(pAktl)in hippocampus tissues was detected by Western blot.Immunohistochemistry was used to detect the expression of insulin receptor substrate 1(IRS1),glucose transporter 4(GLUT4)in hippocampus tissues.Results The hippocampus of db/db mice and APP/PS1 mice showed pathological changes such as disorganization of neuronal arrangement,significant reduction of cell number;glucose content was elevated in both mice(P<0.05);hippocampus insulin was elevated in db/db mice(P<0.01)and decreased in APP/PS1 mice(P<0.01);the protein expressions of PI3K,pAktl/Aktl,CLUT4 were increased in both mice(P<0.01),and IRS1 protein expression was significantly elevated(P<O.01).After the intervention of Dihuang Yinzi,the hippocampus patho

关 键 词：糖尿病脑病 阿尔茨海默病 异病同治 胰岛素抵抗 地黄饮子 

分 类 号：R259[医药卫生—中西医结合]