辛开苦降方对初发2型糖尿病KKay小鼠肝脏胰岛素抵抗及IRS-2/PI3K通路的影响（英文）  

Effects of Xinkai Kujiang Formula on hepatic insulin resistance and the IRS-2/PI3K pathway in KKay mice with incipient type 2 diabetes mellitus

作　　者：张先慧[1] 胡照娟[1] 张艳红[1] 张冬梅[1] 娄利霞[1] 柳红芳[1] 

机构地区：[1]北京中医药大学东直门医院,北京100700

出　　处：《中华中医药杂志》2015年第5期1774-1779,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：National Natural Science Foundation of China(No.81072735)~~

摘　　要：目的:研究辛开苦降方中药对KKay 2型糖尿病(T2DM)小鼠肝脏胰岛素抵抗及胰岛素受体底物-2/磷脂酰肌醇3-激酶通路(IRS-2/PI3K)的影响,探讨其改善肝胰岛素抵抗(IR)的分子机制。方法:将T2DM KKay小鼠按血糖轻重程度分层随机分为4组,即模型组,辛开苦降组、辛开苦降大剂量组、罗格列酮组。正常组选10周龄雄性C57BL/6J小鼠10只。正常组及模型组给予0.5%羧甲基纤维素钠溶液(CMC)灌胃。各组连续灌胃给药4周后取材,采用葡萄糖氧化酶法测血浆葡萄糖(FPG),放射免疫分析法测血浆胰岛素浓度(FINS),免疫蛋白质印迹法测定肝组织IRS-2、磷酸化胰岛素受体底物-2(p-IRS-2)、PI3K、磷酸化磷脂酰肌醇3-激酶(p-PI3K)、胰岛素受体底物-3(IRS-3)、胰岛素受体底物-4(IRS-4)蛋白表达水平,实时荧光定量PCR法测定肝组织PI3K、IRS-2、IRS-3、胰岛素受体(Ins R)的m RNA表达水平。结果:与正常组相比,模型组胰岛素敏感指数明显降低(P<0.01),FPG、FINS均明显升高(P<0.01)。辛开苦降组及其大剂量组两组FINS、胰岛素敏感指数均高于模型组(P<0.01,P<0.05)。免疫蛋白质印迹结果显示:与正常组相比,模型组小鼠肝脏IRS-2、p-IRS-2、PI3K、p-PI3K蛋白表达均明显减少(P<0.01),IRS-3、IRS-4蛋白表达与正常组比较明显增多(P<0.01)。辛开苦降组和辛开苦降大剂量组与模型组相比,小鼠肝脏IRS-2、p-IRS-2、PI3K、p-PI3K蛋白表达均明显增多(P<0.01,P<0.05),IRS-3、IRS-4蛋白表达明显减少(P<0.05),辛开苦降组和辛开苦降大剂量组两组之间无明显统计学差异。实时荧光定量PCR结果显示:与正常组相比,模型组小鼠肝组织PI3-K、IRS-2、IRS-3及Ins R m RNA表达均降低(P<0.01)。与模型组相比,辛开苦降组PI3K m RNA表达升高(P<0.05)。结论:辛开苦降方可改善KKay T2DM小鼠的IR,增加胰岛素的敏感性,调节KKay小鼠肝脏IRS-2、p-IRS-2、PI3K、p-PI3K、IRS-3、IRS-4蛋白和PI3K m RNA表达水平,提示�Objective： To investigate the effects of Xinkai Kujiang Extract Formula （XKF） on hepatic insulin resistance and the insulin receptor substrate 2 （IRS-2）/phosphatidylinositol 3-kinase （PI3K） pathway in KKay mice with type 2 diabetes mellitas （T2DM） meUitus, and determine the molecular mechanism involved in its improvements of insulin resistance. Methods： KKay mice with T2DM were randomly assigned into four groups according to blood glucose levels： diabetic model group, XKF group, high-dose XKF group, and Rosiglitazone group. Ten male C57BL/6J mice （10 weeks old） served as a non-diabetic control group. The mice in the control and model groups were intragastrically administered 0.5% sodium carboxymethyl cellulose solution. Blood and tissue samples were collected after 4 weeks of treatment. Fasting plasma glucose levels were measured using the glucose oxidase method. Fasting plasma insulin concentrations were determined by radioimmunoassay. Hepatic IRS- 2, phosphorylated （p） insulin receptor substrate 2 （p-IRS-2）, PI3K, p-PI3K, IRS-3, and IRS-4 protein expression levels were determined by Western blotting. Hepatic PI3K, IRS-2, IRS-3, and insulin receptor （IR） mRNA expression levels were determined by real-time fluorescent quantitative PCR. Results： The insulin sensitivity index was significantly lower （P〈0.01）, but fasting blood glucose and serum insulin levels were significantly higher （P〈0.01） in the model group than in the control group. Serum insulin levels （P〈0.01） and the insulin sensitivity index （P〈0.05） were significantly higher in the high-dose XKF groups than in the model group. Western blotting revealed that hepatic IRS-2, p-IRS-2, PI3K, and p-PI3K protein expression was significantly lower （P〈0.01）, whereas IRS-3 and IRS-4 protein expression levels were significantly higher in the model group （P〈0.01） than in the control group. Hepatic IRS-2, p-IRS-2, PI3K, and p-PI3K protein expression levels were significandy higher in the low-

关 键 词：辛开苦降方 2型糖尿病 KKAY小鼠 肝胰岛素抵抗 IRS-2/PI3K通路 

分 类 号：R285.5[医药卫生—中药学]