益气化浊胶囊通过AMPK信号通路改善胰岛素抵抗的实验研究

Experiment Research about Effects of Yiqi Huazhuo Capsule on IR through AMPK Signal Channel

目的:通过观察益气化浊胶囊对自发性2型糖尿病KKAy小鼠骨骼肌中腺苷酸活化蛋白激酶(AMPK)蛋白水平、葡萄糖转运蛋白4(GLUT-4)蛋白水平、脂肪组织中丙二酸单酰辅酶A脱羧酶(MLYCD)活性表达情况的影响,探讨其改善胰岛素抵抗的作用机制。方法:选用与人类2型糖尿病(T2DM)发病机制相似的自发型2型糖尿病KKAy小鼠作为动物模型,喂以高脂饲料。以益气化浊胶囊高剂量[2 g/(kg·d)]、低剂量[1g/(kg·d)]灌胃动物,设吡格列酮[2.5 mg/(kg·d]为阳性对照组,空白对照组选取与KKAy小鼠同源的C57BL/6J小鼠,喂以普通饲料。给药10周后处死小鼠,取骨骼肌组织,用western-blot法检测GLUT-4蛋白表达水平、p-AMPK蛋白表达水平,用ELISA法测定MLYCD活性。结果:与空白对照组相比,模型组中骨骼肌p-AMPK、GLUT-4蛋白表达水平、脂肪MLYCD活性显著降低;与模型对照组相比,益气化浊胶囊高、低剂量组、阳性对照组p-AMPK、GLUT-4蛋白表达水平、MLYCD活性升高。结论:益气化浊胶囊能够改善KKAy小鼠胰岛素抵抗情况,其作用机制之一可能与其激活骨骼肌中AMPK信号通路,从而上调GLUT-4蛋白含量、增加MLYCD活性有关。

Objective ：To observe the effects of Yiqi Huazhuo Capusle on AMPK and GLUT- 4 expressions of skeletal muscle and the activity of MLYCD in fat, and to explore its possible molecular mechanism in increasing insulin sensitivi- ty. Methods： The spontaneous type 2 diabetes KKAy mice which are similar with the human type 2 diabetes pathogenesis were as animal models, feeding high fat diet. Yiqi Huazhuo Capsule of high - dose [ 2 g/（ kg · d） ], low - dose [ I g/（ kg · d） ] were used to treat KKAy mice. Positive control group was treated with pioglitazone [ 2.5 rag/（ kg · d） ], blank controller was the C57BL/6J mouse and homologous of KKAy mouse, feeding by common forage. After 10 weeks, the mice were sacrificed to take skeletal muscle and fat. The Western - blot method was used to detect the protein of GLUT - 4,p -AMPK and ELISA was to detect the activity of MLYCD. Results：As compared with normal control group, the ex- pressions of GLUT-4,p- AMPK and MLYCD in the model set were reduced markedly. As compared with model con- trol, the expressions of GLUT- 4, p- AMPK and MLYCD in the Yiqi Huazhuo Capsule of high- dose group,low -dose group and positive control group had been risen in varying degrees. Conclusion： Yiqi Huazhuo Capsule improves FPG and insulin resistance （IR） in KKAy mice and one of its mechanism is probably related to raising expression levels of GLUT - 4 and AMPK protein in skeletal muscle and the activity of MLYCD in fat, and enhancing the transport of glucose in pe- ripheral tissue.