基于代谢组学的卫气节律机制及方药干预研究

The theory of wei-energy is an important part of TCM.In Neijing, there are lots of discussions on the function and running feature of wei-energy.The key point of modern study lies in material classification of wei-energy and its range.Recent studies of wei-energy often center on its material basis from the perspective of certain unique function. Therefore, these studies deviate from the concept of integrety in TCM and the theory of Wei-energy in Neijing. Basing on the theory of integrety and the original meanining of wei-energy as well as the regular running rules of wei-energy,the present research team make metabolism analysis on normal persons'blood serum and animal's thoracic gland at different times and make contrast on the wei-energy-deficiency patients before and after their treatment.The present research team makes use of metabolic skills to recognize standard substance of metabolism that has obvious changes. The present study involves the path of metabolism and its changing ruls, which illustrate the change of wei-energy accordance with time and the corresponding mechanism between wei-energy and the disturbance of the traditional Chinese medicine. Furthermore, the present research team probes into the material basis of wei-energy and its rules for endogenous changes. The present research will reveal the essence or wei-energy, its running scope and its theoretical connotation from a new standard.and it will provide scientific proof for the precaution and treatment of Wei-energy and its related diseases.

卫气理论是中医基础理论的重要组成部分，《内经》对卫气的功能作用及运行特点，已有较为详尽的论述，现代研究的关键在于卫气的物质基础分类及范围界定。目前卫气现代研究大多从单一功能探求其相应物质基础，偏离了中医的整体观念和《内经》卫气理论的原意。本课题在遵循整体原则和《内经》卫气理论本意的基础上，依据卫气年、月、日的周期规律，在不同的时间点对正常人的血清、动物胸腺等进行代谢组学检测，并以临床卫气虚患者治疗前后对比进行反证。运用代谢组学技术，从基础实验和临床验证两个方面识别有显著变化的代谢标准物。研究所涉及代谢途径和变化规律，以阐明卫气随时间变化的规律及中药干预相应机制，进而探求卫气的物质基础和内源性变化规律。通过此项研究将对卫气的本质、卫气的运行范围、卫气的理论内涵有一个崭新水平的展现，也将为临床预防及治疗卫气相关疾病提供新的科学依据。

本课题组突破以往从卫气单一的功能来探求其物质基础的研究框架，采用代谢组学技术来研究伴随卫气节律和药物干预而出现的内源性代谢的变化。通过对卫气节律变化相应的代谢物及卫气虚证患者采用玉屏风颗粒干预前后代谢物的分析，确定伴随卫气节律波动的“代谢组学特征”的小分子标志化合物，进而从基础实验及临床角度多层面验证和筛选与卫气节律相吻合的物质分类及范围界定，为卫气理论及其相关的疾病的诊断和治疗提供新的思路和方法。.本课题的主要结论是：①卫气虚证患者玉屏风颗粒干预的血浆代谢组学研究。疾病组在治疗前和健康组的代谢谱存在明显不同，初步推测出半胱氨酸、L-亮氨酸、L-蛋氨酸、苏氨酸、焦谷氨酸等5种物质为卫气虚证的潜在标志物；经玉屏风颗粒治疗后疾病组的代谢谱有积极改变，向健康组回归；治疗后半胱氨酸、L-蛋氨酸、苏氨酸、焦谷氨酸出现回调，向健康组接近，L-亮氨酸反而进一步偏离健康组。提示：玉屏风颗粒干预后卫气虚证患者出现代谢谱的回归；半胱氨酸、L-蛋氨酸、苏氨酸、焦谷氨酸在治疗后回调，反证了这四种物质作为卫气虚证潜在证候标记物的可靠性；纠正紊乱的氨基酸代谢可能是玉屏风颗粒在分子层面治疗卫气虚证的作用机制之一。②基于代谢组学的卫气虚证年节律的实验研究。春、夏、秋、冬发现卫气虚证的潜在标志物不完全相同，分别为L-天冬氨酸、半胱氨酸、L-蛋氨酸、赖氨酸、高半胱氨酸等。其中半胱氨酸、L-蛋氨酸在四个季节均出现，呈现春夏较强、秋冬较弱的规律，与卫气本身的春夏强、秋冬弱的年节律相符合。提示 ：四季发现不完全相同的证候标志物是因为季节变化刺激机体，使体内的代谢模式或代谢通路随之发生改变，代谢产物产生变化导致，与卫气的节律存在相关性；半胱氨酸、L-蛋氨酸在四个季节均出现，其春夏较强、秋冬较弱的规律符合卫气本身的年节律，推测是伴随卫气年节律的潜在证候标志物。③基于胸腺代谢组学对正常 SD 雄性大鼠卫气年节律变化的研究：春、夏、秋、冬四个季节中正常组与模型组具有明显差异的代谢谱变化，本研究在负离子模式下共获得5个主成分，我们推测出环磷酸腺苷、苹果酸、高肌肽、尿苷、皮质醇为体现卫气在不同季节变化规律的潜在标志物。④对卫气年、月节律变化的代谢潜在标志物进行进一步的研究。