“劳倦过度、房室不节”肾阳虚小鼠模型脑基因芯片研究

Study on brain gene chip of mouse model with def iciency of kidney-yang syndrome

目的:应用基因芯片探讨"劳倦过度、房室不节"肾阳虚小鼠模型脑基因改变,试图说明传统中医建模方法及其肾阳虚表现,力求从基因水平上揭示肾阳虚的本质。方法:采用雄雌鼠比例1:6同笼喂养和雄鼠每日游泳4周,以诱导"劳倦过度、房室不节"肾阳虚鼠模型。用36K mouse genome array鼠脑芯片检测正常对照组和肾阳虚模型组鼠脑基因,并以二者荧光信号相对强度比值≥2和≤0.5筛选差异显著基因,并进一步查阅北京博奥生物分子注释系统,进行基因功能归类。结果:诱导成功"劳倦过度、房室不节"肾阳虚鼠模型,绘制了对照组与模型组基因表达谱比较的散点图。共筛选出186个差异表达基因,其中上调基因150个,下调基因36个。其中共有已知基因98个,上调已知基因71个,下调已知基因27个。下调基因中主要为生长激素、泌乳素、促性腺激素、黑色素和脑脂肪酸结合蛋白(B-FABP)相关基因;上调基因主要为炎症/免疫、促细胞凋亡相关基因以及影响多巴胺生成的限速酶、影响凝血纤溶机制和精子发育代谢的相关基因。结论:通过基因芯片对肾阳虚动物模型的检测,从基因水平上对肾阳虚证的本质做了一些诠释。

Objective： To inquire into the brain gene change of mouse model with kidney-yang deficiency induced by overwork and excess sex intercourse, explain the method of building up animal model according to traditional Chinese medical pathogeny and manifestations of kidney-yang deficiency, the essence of kidney-yang deficiency syndrome can be revealed from gene level. Methods： The model of kidney-yang deficiency was established by each male mouse with six female mouse keeping in a same cage, and all the male mouse were forced to swim for 30-40 minutes everyday lasting for four weeks. The brain genes of mouse in control group and model group were detected with 36K mouse genome array. The differentially expressed genes were screened if the relative fluorescence intensity ratio of the two groups 2 or 0.5 and further classified according to gene function by using Molecule Annotation System （MAS） created by CapitalBio Corp. Beijing, China. Results： The mouse model with kidney-yang deficiency was successfully established by method of overwork and excess sex intercourse, the scatter plot of gene expression profiles of comparing control group with model group was drawn. 186 differentially expressed genes were screened, including 150 up-regulated genes and 36 down-regulated genes. Among the 186 genes only 98 genes were known, including 71 up-regulated genes and 27 down-regulated genes. The down-regulated genes contained genes related to： growth hormone （Gh） ,gonadotropic hormone, prolactin（PRL）, melanin-concentrating hormone （MCH）, and brain fatty acid-binding protein（B-FABP）. The up-regulated genes included genes related to： inflammation/immunization, cell apoptosis, rate-limiting enzyme relative to DA generation, related genes which influence mechanism of blood coagulation, fibrinolysis and development, metabolism of sperm. Conclusion： From detecting the brain gene change of mouse model with kidney-yang deficiency, the essence of kidney-yang deficiency syndrome was explained from gene level.