芍药甘草"酸甘化阴"调节Glu-GABA平衡的"柔筋止痉"中枢作用机制研究

Shaoyao Gancao Tang is a famous formula in Shang Han Lun. It has the functions of norishing blood and yin,pacifying liver,and relieving spasm and pain. It is representative of formulas that has the actions of sour and sweet tranforming into yin and relaxing muscles and relieving spasm. This compatibility is simple and delicate. And its curative effect on spastic paralysis is accurate. On the basis of modern researches of this compatibility combining with knowledge of poststroke spasticity in Chinese and western and guided by "yin-yang balance" theory,we plan to observe the effect of different dose compatibility of Shaoyao and Gancao on neuroethology assessment and pain threshold of post-stroke spasticity rats in order to get the optimum dose and synergism. Then to detect inhibitory and excitatory neurotransmitters content and relevant receptors expression in brain of post-stroke spasticity rats to reveal the cerebral pathological changes and investigate characteristics of the compatibility. Then, we plan to observe the interaction of GABA and NMDA receptors in NMDA-induced excitoxicity cortex cells and neuroprotection by blood serum or cerebrospinal fluid containing Shaoyao and Gancao. We also detect "Src-NR2-nNOS" signal targets comparing with receptor agonist and antagonist. This project will explore the relation of post-stroke spasticity, peripheral neuromuscular function, and central neurotransmitters-receptors system, and mechanism of central action and control targets of compatibility of Shaoyao and Gancao relieving spasm through regulating "Glu-GABA"system balance on molecular level. In addition, it will reveal the characteristics and advantages of compatibility of Shaoyao and Gancao, and reveal science essence of "relaxing muscles and relieving spasm". What's more, it will bring about a better understanding of the Traditional Chinese Medicine theory of five flavours.

芍药甘草汤是《伤寒论》的名方，为酸甘化阴，柔筋止痉的代表方剂，其配伍精当，治疗痉挛性瘫痪疗效确切。课题组在对其配伍现代研究的基础上，结合中西医对脑卒中后痉挛性瘫痪的认识，以中医学"阴阳平衡"理论为指导，采用脑卒中痉挛偏瘫动物模型，观察芍药甘草不同比例的止痉作用，研究两药配伍之后的协同增效作用和最佳剂量配比；并通过观察两药配伍对脑内抑制性神经递质和兴奋性神经递质的含量及其受体的影响，揭示脑卒中痉挛状态"GABA-Glu"系统的病理变化，探讨其酸甘化阴、柔筋止痉的作用特点；进一步采用NMDA兴奋性神经毒细胞模型，借用NMDA受体、GABA受体激动剂和拮抗剂，研究"Src-NR2-nNOS"信号介导的受体之间的相互作用，探讨芍甘配伍调控"GABA-Glu"平衡的分子机制。本课题旨在阐明芍药甘草"酸甘化阴"的配伍特色和优势，揭示中药"柔筋止痉"的现代科学内涵，是中药五味理论的有益探索。

背景：芍药甘草汤是《伤寒论》的名方，为酸甘化阴，柔筋止痉的代表方剂，其配伍精当，治疗痉挛性瘫痪疗效确切。内容：本项目以中医学“阴阳平衡”理论为指导，采用脑卒中痉挛偏瘫动物模型，揭示芍药甘草配伍之后的协同增效作用和最佳剂量配比；从中枢抑制性和兴奋性神经递质与受体角度，研究脑卒中痉挛状态“GABA-Glu”系统的病理变化，探讨芍药甘草柔筋止痉的中枢作用特点；采用NMDA兴奋性神经毒细胞模型，借用受体抑制剂和激动剂，研究“Src-NR2-nNOS”信号介导的受体之间的相互作用，探讨芍甘配伍调控“GABA-Glu”平衡和神经保护的分子机制。结果：实验研究表明，芍药甘草汤对脑卒中后痉挛偏瘫状态有较好的改善作用，能够显著改善神经症状，降低肌张力，改善痛阈，具有解痉止痛之效，其解痉止痛最佳配伍比例为3：1（芍药：甘草）。芍药甘草汤可能是通过增加抑制性氨基酸含量和激活其受体，抑制Src-NR2-nNOS信号通路，抑制兴奋性神经能信号，从而达到神经保护和“柔筋止痉”的作用。意义：本课题将传统中药配伍理论与现代药理学技术相结合，探讨芍药甘草“酸甘化阴”的配伍特色和优势，揭示中药“柔筋止痉”的现代科学内涵，是中药五味理论的有益探索，具有学术价值和临床指导意义。

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