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[基于金匮肾气丸与右归丸对肾阳虚证反证小鼠模型治疗效果的脑基因芯片研究]

[Gene chip study on cerebral gene of effect of Jinkui Shenqiwan and Youguiwan on mouse model of kidney-yang asthenia with syndrome disproved according to therapeutic efficacy of drugs used].

作者信息

Yang Yuhua, Li Zhen, Sun Jing

机构信息

Shandong University of Traditional Chinese Medicine, Jinan 250014, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2009 May;34(9):1124-8.

Abstract

OBJECTIVE

To inquire into the cerebral gene change of effect of Jingui Shenqiwan and Youguiwan for animal model of kidney-yang asthenia caused by excessive physical and sexual activities, which may study the effect mechanism of the medicine with syndrome disproved therapeutic efficacy of drugs used.

METHOD

Male mice of Kunming species, weight 35-40 g, and female weight 28-35 g were randomly divided into four groups: control group, model group and treatment groups of Jigui Shenqiwan and Youguiwan in which there were ten male mice, fifteen ones, ten ones and ten ones, respectively. All of them were fed normally, and poured into their stomach with 0.5 mL of distill water for each mouse in the control group and model group, and with 0.5 mL suspension of the drugs (including 1.1 g x kg(-1) drug) for each one in the treatment groups every day. The mice in the model group and treatment groups were kept by means of each male mouse with six female mice in the same cage, and all male mice swam until they gradully submerged and were scooped up from water once everyday for lasting four weeks to induce the kidney-yang asthenia with excessive physical and sexual activities. Animals' manifestation such as fearing cold, activity and responses, mouse' fur and so on were observed. The brain gene were detected with the mouse brain gene chip of 36K Mouse genome array made by Capital Bio Corp. Beijing, China, and the differential expression gene were screened according to the ratio equal to or above 2 and equal to or below 0.5 with the related fluorescent intensity comparing the two groups, which could be further verified in the light of partly differential expression gene with qRT-PCR.

RESULT

The mouse model of kidney-yang asthenia in the model group was successfully induced by way of excessive physical and sexual activities. There were twenty-three genes among up-regulated genes in the model group versus control group but down-regulated genes in the treatment groups versus model group, chiefly including the genes association with inflammation/immunization, neurotransmissions/ signal transduction and so on. There were six genes among the down-regulated genes in the model group versus control group but upregulated in the treatment groups versus model group, mainly involving the related genes of cellular cycle and structure, neurotransmissions/signal transduction, transcription and et al.

CONCLUSION

Jigui Shenqiwan and Youguiwan may make it markedly up-regulated to notably down-regulated genes of hormone and melanin-concentrating hormone (MCH) for model of mouse of kidney-yang asthenia, and promote cellular proliferation, which can inquire into effect mechanism of the drug in genetic level at the same time.

摘要

目的

探讨金匮肾气丸与右归丸对房劳过度致肾阳虚动物模型脑基因变化的影响,以研究药物的作用机制,为临床药物疗效不佳提供证型方面的研究依据。

方法

选取体重35~40g的雄性昆明种小鼠和体重28~35g的雌性昆明种小鼠,随机分为对照组、模型组、金匮肾气丸治疗组和右归丸治疗组,每组分别为10只、15只、10只、10只。各组小鼠正常饲养,对照组和模型组小鼠每日每只灌胃0.5mL蒸馏水,治疗组小鼠每日每只灌胃0.5mL药物混悬液(含生药1.1g·kg-1)。模型组及各治疗组雄性小鼠按1∶6与雌性小鼠合笼饲养,所有雄性小鼠每日游泳至逐渐下沉后捞出,持续4周,造成房劳过度致肾阳虚动物模型。观察动物的畏寒、活动、反应、毛发光泽等表现。采用北京博奥生物有限公司生产的36K小鼠全基因组表达谱芯片检测脑组织基因表达,以两组相关荧光强度比值≥2或≤0.5为标准筛选差异表达基因,并对部分差异表达基因进行实时荧光定量聚合酶链反应(qRT-PCR)验证。

结果

模型组小鼠通过房劳过度法成功诱导出肾阳虚模型。模型组与对照组比较有23个基因上调,模型组与金匮肾气丸及右归丸治疗组比较以上调基因下调,主要涉及炎症/免疫、神经传递/信号转导等相关基因;模型组与对照组比较有6个基因下调,模型组与金匮肾气丸及右归丸治疗组比较以下调基因上调,主要涉及细胞周期与结构、神经传递/信号转导、转录等相关基因。

结论

金匮肾气丸和右归丸可使肾阳虚小鼠模型中激素及促黑素(MCH)相关基因明显上调至下调,促进细胞增殖,同时从基因水平初步探讨了药物的作用机制。

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