糖痛方对糖尿病周围神经病变模型大鼠坐骨神经PI3K、Akt、mTOR表达的影响

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目的:观察益气活血中药糖痛方对DPN大鼠坐骨神经自噬通路蛋白PI3K、Akt、mTOR表达的影响,探讨其作用机制。方法:雄性SD大鼠60只,随机选取15只作为正常组,其余大鼠经STZ+缺血再灌注方法建立DPN模型,按随机数字表法分为模型组、糖痛方低剂量组、糖痛方高剂量组,每组15只。糖痛方高、低剂量组分别灌胃36.67、18.33 g/kg糖痛方水煎液,1次/d。连续灌胃8周后,检测坐骨神经传导速度,采用qRT-PCR和Western Blot法检测坐骨神经PI3K、Akt、mTOR mRNA及蛋白水平,采用HE染色观察坐骨神经纤维结构。结果:与模型组比较,糖痛方低、高剂量组运动神经传导速度、感觉神经传导速度、肌肉复合动作电位、感觉神经动作电位提高(n P<0.05),坐骨神经PI3K[(6.05±0.18)、(3.36±0.29)比(11.57±1.93)]、Akt[(1.26±0.13)、(0.64±0.04)比(1.86±0.06)]、mTOR[(1.82±0.11)、(0.92±0.06)比(2.68±0.18)]mRNA水平降低(n P<0.05),PI3K[(0.40±0.00)、(0.19±0.02)比(0.61±0.03)]、Akt[(0.64±0.02)、(0.45±0.01)比(0.83±0.02)]、mTOR[(0.17±0.01)、(0.09±0.00)比(0.34±0.01)]蛋白表达降低(n P<0.05);模型组神经纤维松散肿胀,髓鞘变薄,轴突闭锁,糖痛方低、高剂量组神经形态趋于正常,髓鞘及轴突均形态较好。n 结论:糖痛方可改善DPN大鼠坐骨神经的传导速度及电位波幅,改善神经损伤,减轻脱髓鞘改变,改善轴突形态,保护神经纤维结构,其作用机制可能与激活PI3K/Akt/mTOR信号通路抑制细胞过度自噬有关。“,”Objective:To observe the effect of n Tangtong formula on the expression of autophagy pathway proteins PI3K, Akt and mTOR in sciatic nerve of DPN rats, and to explore its mechanism.n Methods:There were 60 male SD rats, 15 of which were randomly selected as the normal group, and the other rats were used to establish DPN model with STZ + ischemia-reperfusion method. Then they were divided into model group, n Tangtong formula low-dose group and n Tangtong formula high-dose group, with 15 rats in each group with random number table method. 36.67 g/kg n Tangtong formula was administered by gavage in the high-dose group and 18.33 g/kg n Tangtong formula was administered by gavage in the low-dose group, once a day. After 8 weeks of continuous gavage, the conduction velocity of sciatic nerve was detected. The mRNA and protein expression levels of PI3K, Akt and mTOR were detected by PCR and Western blot. The structure of sciatic nerve fibers was observed by HE staining.n Results:Compared with the model group, the motor nerve conduction velocity, sensory nerve conduction velocity, muscle compound action potential and sensory nerve action potential in the low-dose n Tangtong formula group and high-dose n Tangtong formula group were increased (n P<0.05). The expression of PI3K mRNA(6.05±0.18, 3.36±0.29n vs. 11.57±1.93), Akt mRNA(1.26±0.13, 0.64±0.04 n vs. 1.86±0.06), mTOR mRNA(1.82±0.11, 0.92±0.06 n vs. 2.68±0.18) of sciatic nerve in rats of the low-dose and high-dose group were increased (n P<0.05). The expression of PI3K(0.40±0.00, 0.19±0.02n vs. 0.61±0.03), Akt(0.64±0.02, 0.45±0.01 n vs. 0.83±0.02), mTOR(0.17±0.01, 0.09±0.00 n vs. 0.34±0.01)of sciatic nerve in rats of the low-dose and high-dose group were increased (n P<0.05). The model group\'s nerve fibers were loose and swollen, myelin sheath became thin, and the axis Atresia, the neuromorphology of the low-dose and high-dose group tended to be normal, and the morphology of myelin sheath and axon were better.n Conclusions:Tangtong formula could improve the conduction velocity and potential amplitude of sciatic nerve in DPN rats, reduce nerve injury and demyelinating changes, improve axon morphology and protect nerve fiber structure. Its mechanism might be related to activating PI3K/Akt/mTOR signal pathway and inhibiting excessive autophagy.n
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