温补肾脾方药对肾脾阳虚模型大鼠Akt信号通路的影响  

Effect of Recipe of Warming and Tonifying Spleen and Kidney on Protein Kinase B(PKB-Akt)Signalling Pathway in Deficiency of Kidney and Spleen-Yang Model Mice

作　　者：周艳艳[1] 郭煜晖[2] 周安方[1] 徐安莉[1] 赵敏[1] 

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]三峡大学,湖北宜昌443002

出　　处：《中华中医药学刊》2013年第11期2447-2449,I0004,I0005,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81102529);湖北省教育厅科研项目(Q20112011)

摘　　要：目的:通过采用肌注氢化可的松和灌胃大黄水煎液复合方法复制肾脾阳虚模型大鼠,观察温补肾脾方药对肾脾阳虚模型大鼠大脑海马组织形态及Akt/FOXO3a信号通路的影响,探讨肾脾阳虚模型大鼠衰老的可能机制。方法:40只SD大鼠,随机分为正常对照组、模型组、维生素E组和桂附理中丸组。正常对照组大鼠肌肉注射生理盐水,同时灌胃生理盐水,其余组大鼠肌肉注射氢化可的松,同时灌胃大黄水煎液,维生素E组灌胃维生素E混悬液,桂附理中丸组灌胃桂附理中丸混悬液,共30 d。取其大脑海马组织,透射电镜下观测海马CA1区超微结构的变化。Westernblot法和免疫组织化学染色方法检测Akf、FOXO3a蛋白表达。结果:肾脾阳虚模型大鼠大脑海马CA1区细胞器及神经元出现衰老表现,桂附理中丸组大鼠海马CA1区细胞轮廓清晰,无细胞器及神经元细胞衰老征象,用药效果明显优于维生素E组。与正常对照组比较,模型组大鼠大脑海马磷酸化Akt蛋白表达明显升高(P<0.05>,磷酸化FOXO 3a蛋白表达显著升高(P<0.01);与模型组比较,桂附理中丸组大鼠大脑海马磷酸化Akt蛋白明显降低(P<0.05),磷酸化FOXO 3a蛋白显著降低(P<0.01)。结论:肾脾阳虚模型大鼠大脑海马细胞衰老的病理改变明显,肾脾阳虚可导致模型大鼠磷酸化的Akt、FOXO3a蛋白表达增强,降低机体的抗氧化能力,从而导致衰老。温补肾脾方药可以抑制Akt、FOXO3a蛋白的磷酸化,增强抗氧化酶活力,改善大脑海马组织神经元细胞的病理改变,发挥延缓衰老的作用。Objective ： Using injected with hydrocortisone and administered with decoction liquid of Dahuang had estab- lished deficiency of kidney and spleen - Yang disease models. The hippocam - pus changes and the influence of the pro- tein Akt/FOXO3a signalling pathway were observed. Methods： Totally 40 SD male mice at the age of eight weeks were randomly divided into normal control group, model group, Vitamin E group, Guifu Lizhong Pill group, ten normal mice in each group, which were separately given medicine. Excluding normal control group, the mice were injected with hydro- cortisone and administered with decoction liquid of Dahuang for established deficiency of kidney and spleen - Yang dis- ease models, the procedures lasted for 30 days. The ultrastructure of hippocam - pus changes were observed and adopting protein immune imprinting （ Western blotting） method and immune histochemistry staining method were used to detections of Akt, Foxo3a protein expressions. Results： Compared with the normal control group, the model group mice hippocampal organization organelles were reduced, the neuron atrophy, neurons synaptic gap widening; compared with the model con- trol group, the Vitamin E and Guifu Lizhong Pill group mice hippocampal organization organelles were increased, withers as rarely as the neuron atrophy, neurons synaptic gap widening. Compared with the normal control group, the model group mice P- Akt positive cells were obviously increased, and the average gray level value was significant（P 〈0.05 ） , P- Foxo3a positive cells were significantly increased, and the average gray level value was significant（P 〈0.01 ） ; com- pared with the model control group, the Guifu Lizhong Pill group mice P- Akt positive cells had obviously reduced（ P 〈 0.05）, P- Foxo3a positive cells had significantly reduced（ P 〈 0. 01 ）. Conclusion： The expression of phosphorylated Akt and foxo3a protein of deficiency of kidney and spleen - Yang disease models mice brain hippocampus were increased, resulting

关 键 词：肾脾阳虚 衰老 Akt FOXO3A 温补肾脾方 

分 类 号：R285.5[医药卫生—中药学]