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目的研究中药降糖复方水提取物(WEJTD)对自发性2型糖尿病(T2DM)模型KK-Ay小鼠骨骼肌糖代谢PI3K/Akt信号通路的影响。方法将KK-Ay小鼠按随机数法随机分为5组:模型组、盐酸二甲双胍(MH,250 mg/kg)组和WEJTD低、中、高剂量(2、4、8 g/kg)组,C57BL/6J小鼠作为对照组,每天ig给药1次,共12周,对照组和模型组ig等体积蒸馏水。给药12周后,取小鼠血清,ELISA试剂盒法检测胰岛素水平;Trizol法提取肌肉组织中的RNA,荧光实时定量PCR(q RT-PCR)检测磷脂酰肌醇激酶-3(PI3K)、蛋白激酶B(Akt)、葡萄糖转运蛋白-4(GLUT-4)、糖原合成酶激酶-3β(GSK-3β)、糖原合成酶(GS)、胰岛素受体底物(IRS-1)m RNA水平。结果与模型组比较,WEJTD高、中剂量显著上调小鼠血清胰岛素水平、骨骼肌中IRS-1 m RNA水平(P<0.05、0.001);高、中、低剂量均显著下调骨骼肌中GSK-3βm RNA水平,显著上调PI3K、Akt、GLUT-4、GS m RNA水平(P<0.05、0.01、0.001)。结论中药降糖复方通过上调PI3K/Akt信号通路,减少糖原沉积,刺激骨骼肌中葡萄糖转运。
Objective To investigate the effects of Chinese herbal compound glucose extract (WEJTD) on the glucose metabolism of PI3K / Akt pathway in skeletal muscle of KK-Ay mouse model of spontaneous type 2 diabetes mellitus (T2DM). Methods KK-Ay mice were randomly divided into 5 groups: model group, metformin hydrochloride (MH, 250 mg / kg) and WEJTD low, medium and high dose (2,4,8 g / kg) , C57BL / 6J mice served as a control group, once a day ig administration for 12 weeks, the control group and model group ig equal volume of distilled water. After 12 weeks of administration, the serum of mice was taken and the level of insulin was detected by ELISA kit. The RNA of muscle tissue was extracted by Trizol method and the activity of PI3K was detected by real-time quantitative PCR (q RT-PCR) Protein kinase B (Akt), glucose transporter-4 (GLUT-4), glycogen synthase kinase-3β (GSK-3β), glycogen synthase (GS), insulin receptor substrate (IRS- RNA level. Results Compared with the model group, high and medium dose of WEJTD significantly increased serum insulin and the level of IRS-1 m RNA in skeletal muscle (P <0.05,0.001). High, medium and low doses of WEJTD significantly decreased GSK- 3βmRNA, and significantly upregulated the levels of PI3K, Akt, GLUT-4 and GS m RNA (P <0.05, 0.01, 0.001). Conclusion Traditional Chinese Medicine Hypoglycemic Compound can up-regulate PI3K / Akt signaling pathway, reduce glycogen deposition and stimulate glucose transport in skeletal muscle.