肾脾阳虚对实验大鼠脑组织Klotho基因表达的影响  

The Influence of the deficiency of kidney and spleen-Yang on Klotho gene expression in 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：《时珍国医国药》2013年第12期3060-3063,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81102529)

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,观察肾脾阳虚对模型大鼠大脑海马组织形态及Klotho基因表达的影响,探讨肾脾阳虚模型大鼠衰老的机制。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。透射电镜下观测海马CA1区超微结构的变化。取大脑海马组织实时荧光定量PCR检测Klotho mRNA的表达,Westernblot法和免疫组织化学染色方法检测Klotho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,模型组大脑海马Klotho表达水平明显降低(P<0.05);与模型组比较,桂附理中丸组大鼠Klotho表达水平明显升高(P<0.05)。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,表明肾脾阳虚可导致大鼠衰老。肾脾阳虚可导致模型大鼠大脑海马Klotho表达水平下降,温补肾脾方药可上调Klotho的表达,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。Klotho蛋白低表达是肾脾阳虚模型大鼠衰老的可能机制。Objective Using injected with hydrocortisone and administered with decoction liquid of Rhubarb had established defi- ciency of kidney and spleen - Yang disease models to observe the hippocampus changes and the influence on Klotho gene expres- sion. Methods Totally 40 SD male mice at the age of eight weeks, were randomly divided into normal control group, model group, Vitamin E group and Guifulizhongwan group,with ten normal mice in each group, which were separate｜y given medicine. Excluding normal control group, the mice were injected with hydroeortisone and administered with decoction liquid of Rhubarb for established deficiency of kidney and spleen - Yang disease models, the procedures lasted for 30 days. Observation the uhrastruc- ture of hippocampus changes ; detection of Klotho mRNA expression by real - time fluorescence quantitative polymerase chain reac- tion （PCR） method; adopting protein immune imprinting （Western blotting） method and immune histochemistry staining method to detection of klotho protein expressions. Results Compared with the normal control group, the model group mice which of hipp- ocampal organization organelles were reduced, the neuron atrophy, neurons synaptic gap widening; compared with the model con- trol group, the Vitamin E and Guifulizhongwan group mice which of 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 Klotho expression had obviously decreased（ P 〈 0.05 ） ;compared with the model control group, the Guifulizhongwan group mice whice of Klotho expression levels had obviously increased （ P 〈 0.05 ）. Conclusion The deficiency of kidney and spleen - Yang disease models mice of the brain hippocampus of Klotho expression levels were decreased, resulting in ability to clear the oxygen free radical was reduced, increasing in superoxide damage to the body, to speed up the bodyg aging. The recipe of warming and tonifying the sple

关 键 词：肾脾阳虚 衰老 KLOTHO 大脑海马 

分 类 号：R285.5[医药卫生—中药学]