肾脾阳虚对实验大鼠脑组织Klotho基因表达的影响

发布时间：2017-12-30 22:19

本文关键词：肾脾阳虚对实验大鼠脑组织Klotho基因表达的影响　出处：《时珍国医国药》2013年12期 　论文类型：期刊论文

【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(P0.05);与模型组比较,桂附理中丸组大鼠Klotho表达水平明显升高(P0.05)。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。
[Abstract]:Objective to observe the effect of deficiency of kidney and spleen yang on the morphology of hippocampus and the expression of Klotho gene in rat model by intramuscular injection of hydrocortisone and decoction of rhubarb. Methods Forty SD rats were randomly divided into normal control group, model group, vitamin E group and Gui Fu Lizhong pill group. The rats in other groups were given hydrocortisone intramuscularly, rhubarb decoction, vitamin E suspension in vitamin E group and Gui Fu Li Zhong pill suspension in stomach. The ultrastructural changes of hippocampal CA1 were observed under transmission electron microscope for 30 days. The expression of Klotho mRNA was detected by real-time fluorescence quantitative PCR. Westernblot and immunohistochemical staining were used to detect the expression of Klotho protein. Results the senescence of CA1 organelles and neurons in hippocampus of rats with deficiency of kidney and spleen yang were observed. Gui-fulizhong pills group had clear outline of CA1 cells in hippocampus, no organelle and senescence signs of neuron cells, and the effect of medication was better than that of vitamin E group, compared with the normal control group. In the model group, the expression of Klotho in hippocampus decreased significantly (P 0.05). Compared with the model group, the expression of Klotho in Gui Fu Li Zhong Wan group was significantly higher than that in the model group. Conclusion the pathological changes of aging of hippocampal cells in rats with deficiency of kidney and spleen yang are obvious. The results indicated that deficiency of kidney and spleen yang could lead to aging, deficiency of kidney spleen yang could lead to the decrease of Klotho expression in hippocampus of model rats, and the prescription of warming kidney and spleen could up-regulate the expression of Klotho. To improve the pathological changes of neuron cells in hippocampus and to play the role of delaying senescence. The low expression of Klotho protein is the possible mechanism of aging in rats with deficiency of kidney and spleen yang.
【作者单位】：湖北中医药大学;三峡大学;
【基金】：国家自然科学基金(No.81102529)
【分类号】：R-332
【正文快照】： Klotho基因是Kuro等[1]在1997年研究自发性高血压发现与衰老密切相关的一种基因,主要在脑脉络丛、海马、肾等组织中表达。Klotho基因在小鼠中表达缺失,会出现类似于人类衰老的各种表型,如动脉硬化、骨质疏松、肺气肿、寿命缩短、皮肤萎缩、不育症、运动失调等[2~8]。Yamamoto

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