以药测证绘制肾虚证两大基因网络调控路线图谱

目的 研究肾虚证基因网络调控的路线和规律.方法 选择青年组（4月龄）、老年组（24月龄）、淫羊藿总黄酮（Epimedium Flavonoids,EF）治疗组SD大鼠,取全面反映肾虚的7个组织下丘脑、垂体、肾上腺、淋巴细胞、骨骼、肝脏、肾脏,采用Affymetrix公司的大鼠全基因组芯片再次重复肾虚证基因表达谱差异研究.结果 老年大鼠下丘脑、垂体、肾上腺（HPA轴）显示神经递质γ氨基丁酸（GABA）,促性腺激素释放激素（GnRH）,促甲状腺激素释放激素（TRH）与促甲状腺激素（TSH）,生长激素释放激素受体（GHRH）、胰岛素样生长因子及结合蛋白（IGF、IGFBP）;淋巴细胞显示与细胞生长相关的生长因子相关蛋白（GFRP）,与免疫调节相关的IFNγ、IL-4、IL-6等;骨骼显示甲状旁腺（PTH）、降钙素（calcitonin）与骨基质相关的前胶原、胶原、结缔组织生长因子等;肝脏显示氧化磷酸化相关的细胞色素Cytochrome P450、NADH脱氢酶,与蛋白质代谢相关的谷氨酸脱氢酶（Glutamate dehydrogenase）,与糖代谢相关的葡萄糖-6-磷酸酶（G-6-Phosphatase）均为低表达.EF作用后,可见老年大鼠7个标本低表达的基因几乎全部上调,此外还有大量与衰老机理相关的基因明显上调.包括HPA轴的神经递质如GABA,激素如GnRH,细胞周期相关基因如CyclinB,免疫调节相关基因如NF-κB105;淋巴细胞与生长调节相关的基因如GFRP,免疫调节相关的基因如IFNγ;骨骼的甲状旁腺、降钙素、IGF、前胶原、胶原、GnRH、Progesterone等;肝脏的大量代谢相关的基因如Cytochrome P450、NADH脱氢酶;肾脏的代谢相关基因及钠、氯通道等基因.结论 通过以药测证,发现肾虚证大鼠模型存在神经-内分泌-免疫以及神经-内分泌-骨代谢两大基因调控路线的紊乱,补肾能纠正该网络功能低下.

Syndrome Differentiation through Drug Effects in Mapping the Two Regulatory Pathways of Gene Networks in Shen Deficiency Syndrome

OBJECTIVE: To study the regulatory pathways and rules of the gene networks in Shen deficiency syndrome. METHODS: Tissues of hypothalamus, pituitary, adrenal, lymphocyte, bone, liver and kidney were taken as samples from 4 months' and 24 months' old SD rats and rats after treatment with Epimedium flavonoids (EF), differences of gene expression profile in Shen deficiency syndrome were studied repeatedly with gene chip rat expression set U230 2.0 array from Affymetrix Co. RESULTS: Gene expressions in the aged rats all decreased including neurotransmitter of gamma-aminobutyric acid (gammaGABA), gonadotropin releasing hormone (GnRH), thyrotropin-releasing hormone (TRH), thyroid stimulating hormone (TSH), growth hormone-releasing hormone receptor (GHRH), insulin-like growth factor (IGF) and binding proteins (IGFBP) in hypothalamus, pituitary and adrenal (HPA axis), cell growth-related gene, growth factor related protein, and immune regulatory genes such as interferon gamma (IFN-gamma), interleukin 4 (IL-4) and interleukin 6 (IL-6) in lymphocytes, parathyroid hormone (PTH), calcitonin, procollagen, collagen, connective tissue growth factor in bone, and oxidative phosphorylation genes such as cytochrome P450 and NADH dehydrogenase, glutamate dehydrogenase related with protein metabolism, and glucose-6-phosphatase related with glucose metabolism in liver, most of which were up-regulated after treatment with EF as well as genes related with ageing and cell cycle, such as cyclin B, metabolism related genes and proteins of sodium and chloride channel in kidney. CONCLUSION: Dysfunction of the two regulatory pathways of gene networks as nerve-endocrine-immunity and nerve-endocrine-bone metabolism exists in Shen deficiency syndrome differentiated by effects of drugs, which could be improved by strengthening Shen therapy.