基于阴阳寤寐学说的交泰丸及其拆方对睡眠时相的影响及调节GABA信号通路的作用

In the process of transformation of Yin and Yang in the human body, kidney-yin go up to heart-yang ,heart-yang decline to kidney-yin,heart-yang and kidney-yin must be balanced.It is sleep’s important regulating mechanism.People may suffer from insomnia If they are imbalanced.On the basis of the research of sleeping and wakeing theory(including the national natural science fund of 81173149 and 8122845),we choose Jiaotai pill,which can cure insomnia of imbalance between heart-yang and kidney-yin,for the treatment of insomnia as a research subject. Jiaotai Pill is quite good effect to cure insomnia of imbalance between heart-yang and kidney-yin,however, the biological mechanism is not clear. Our hypothesis is that Jiaotai pill is by adjusting the GABA signaling pathway to improve insomnia. Under the guidanc of TCM theory, the subject research will use universal model of sleep research field - by the method of a small platform in water environment,observe the sleep phase of insomnia rats,hippocampal γ- aminobutyric acid receptor expression levels change with related experiments,detect glutamate decarboxylase, γ- aminobutyrate acid transaminase and γ- aminobutyric acid transporter expression levels.We aim to explore the probable mechanisms of jiaotai pill to improve sleep,and provide objective evidence in the clinical treatment, prevention and health care for jiaotai pills.

在人体内阴阳转换过程中，肾水上济于心、心火下降于肾保持动态平衡为心肾相交、水火既济，是睡眠（寐）与觉醒（寤）的重要调节机制。若阴阳失调形成水火不济，引起心肾不交而导致失眠。该项目在本课题组前期阴阳寤寐学说系列研究（包括国家自然科学基金81173149，81202845等）基础上，选用调节阴阳水火治疗失眠的代表方交泰丸作为研究的主体。交泰丸改善心肾不交型失眠颇收良效，但其生物学机制尚不明确。我们假定，交泰丸是通过调节GABA信号通路来改善失眠。拟采用目前睡眠研究领域普遍认可的模型—小平台水环境法造模，观察交泰丸改善大鼠的睡眠时相，用相关实验观察海马γ－氨基丁酸受体表达水平变化，并检测谷氨酸脱羧酶、γ－氨基丁酸转氨酶及γ－氨基丁酸转运体表达水平的变化等，探索交泰丸改善睡眠的可能机制，进一步阐明阴阳寤寐学说的真谛，以期为本方法在临床治疗、预防保健中提供客观证据。

阴阳寤寐学说中阴主静，阳主动与现代医学γ-氨基丁酸（GABA，属阴）抑制功能，谷氨酸（Glu，属阳）兴奋功能类似，由此相应调节寐与寤的交接。交泰丸使心肾相交，水火相济，则阴阳交感互藏，促进睡眠。本课题采用睡眠剥夺大鼠模型，给予交泰丸、GABAA和GABAB受体激动剂或抑制剂单独或联合治疗，结合高效液相法（HPLC）检测血液中的Glu含量。研究结果发现：交泰丸显著降低失眠模型大鼠血液中的Glu含量，与GABA受体抑制剂联用以后该作用被抑制。课题进一步利用定量PCR、western-blot技术检测大鼠下丘脑GABA信号通路相关基因和蛋白的表达变化，分析交泰丸对该通路的作用机制。研究结果显示：交泰丸可以增加促睡眠物质GABA受体mRNA（GABAA mRNA、GABAB mRNA）表达量和蛋白（GABAA蛋白、GABAB蛋白）表达量，降低促觉醒相关的GAT3蛋白表达量、GAT1蛋白表达量、GABA-T蛋白表达量，增加促睡眠相关的GAD蛋白表达量，改善失眠状态，促进睡眠。以上实验结果表明：基于阴阳寤寐学说的交泰丸改善失眠的机制可能与调节GABA信号通路有关。交泰丸联合GABA受体激动剂后，相关指标与单纯用交泰丸组呈相同方向变化且更加明显，促眠作用明显增强；交泰丸联合GABA受体抑制剂后，相关指标与单纯用交泰丸组呈相反方向变化，促眠作用减弱。交泰丸可能有类似GABA受体激动剂的效应。