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...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,提示调整肝脏的IRS2/PI3K通路相关的蛋白表达可能是辛开苦降方改善KKay T2DM小鼠的IR作用靶点,且单纯加大剂量并对相关蛋白及基因表达影响不大。展开更多
The two major pathogeneses of type 2 diabetes mellitus (T2DM) are insulin resistance and insulin secretion deficiency. During recent years, researches on the molecular target sites of insulin resistance and the mechan...The two major pathogeneses of type 2 diabetes mellitus (T2DM) are insulin resistance and insulin secretion deficiency. During recent years, researches on the molecular target sites of insulin resistance and the mechanism of the signal transduction has made great progress: especially, the study of insulin receptor substrate-2 (IRS-2). Human IRS-2 gene is located at 13q8.6. IRS-2G1057D is a replacement of G (glycine) by D (aspartic acid) at site 1057 of insulin receptor substrate-2, which is caused by simple nucleotide polymorphism. The role of this variant is still not clear. We detected IRS-2G1057D variant in Han population in Liaoning Province by measuring body mass index (BMI), waistline/hip ratio (WHR) and other parameters of insulin secretion, as well as insulin action to explore the relationship between IRS-2G1057D variant and T2DM.展开更多
基金National Natural Science Foundation of China(No.81072735)~~
文摘目的: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,提示调整肝脏的IRS2/PI3K通路相关的蛋白表达可能是辛开苦降方改善KKay T2DM小鼠的IR作用靶点,且单纯加大剂量并对相关蛋白及基因表达影响不大。
文摘The two major pathogeneses of type 2 diabetes mellitus (T2DM) are insulin resistance and insulin secretion deficiency. During recent years, researches on the molecular target sites of insulin resistance and the mechanism of the signal transduction has made great progress: especially, the study of insulin receptor substrate-2 (IRS-2). Human IRS-2 gene is located at 13q8.6. IRS-2G1057D is a replacement of G (glycine) by D (aspartic acid) at site 1057 of insulin receptor substrate-2, which is caused by simple nucleotide polymorphism. The role of this variant is still not clear. We detected IRS-2G1057D variant in Han population in Liaoning Province by measuring body mass index (BMI), waistline/hip ratio (WHR) and other parameters of insulin secretion, as well as insulin action to explore the relationship between IRS-2G1057D variant and T2DM.
