基于整合生物标志物体系的补肾中药延缓衰老作用基础研究

A research stratety "An Integrated Biomarker System to be used to character the ageing process quantificationally on the basis of Quantitative Metabonomic Platform Technology" was proposed here. The research mode integrating "macroscopic mode and microcosmic mode, and combining the overall characterization and the local feature" would be applied here. The holistic metaobolomics character should be described with metabolic profiling analysis,which was used to find out the key pathway related to the ageing process. And the contents of main endogenous metabolites from phospholipid pathway, fatty acid pathway,acylcarnitines,amino acids and biogenic acid pathways were determined by target metabolomics analysis.And integrating the results of metabolomics profiles and target metabolomics analysis, the key metabolic biomarkers would be focused, and the "Integrated Biomarker System" would be constructed by means of artificial neural network, together with other acknowledged ageing indicators, which can be used to character the ageing process quantificationally. And then the "Integrated Biomarker System" would be used to evaluate the effect of kindey-tonifying traditional Chinese medicine on slowing the ageing process. Refer to the chemical material basis studies on Liuwei Dihuang wan and Jinkui Shenqi wan,as well as the results of network pharmacology, the differences on the effective basis of nourishing kidney and warming kidney would be compared. The research may provide a meaningful reference for modern TCM research.

本项目提出"基于定量代谢组学获取定量表征衰老过程的整合生物标志物体系"的研究策略，采用宏观模式与微观模式相结合，整体表征与局部特征相结合的研究模式：通过代谢轮廓谱分析从全景模式研究衰老对内源性代谢物组终端表象的影响，发现与衰老过程密切相关的代谢循环；进行靶标代谢组学研究，通过具体定量研究从特写模式重点关注衰老对磷脂类、脂肪酸、酰基肉毒碱类、氨基酸类、生物胺类等代谢循环主要代谢物水平的影响；整合代谢轮廓谱分析和靶标代谢组学的研究结果，对重点关注的代谢生物标志物进行聚焦；整合定量代谢组学研究与公认的衰老评价指标（量化衰老标准评分、端粒酶活性、超氧化物歧化酶、丙二醛）测定结果，建立定量表征衰老过程的整合生物标志物体系，用于补肾中药延缓衰老作用评价。结合六味地黄丸和金匮肾气丸的化学物质基础和网络药理学研究，阐释"滋补肾阴"与"温补肾阳"延缓衰老作用基础的异同，为中药复方现代研究提供方法借鉴。

本研究旨在将代谢轮廓谱分析与特定代谢循环的靶标代谢组学分析相结合，发现可定性定量表征与衰老过程相关的整合生物标志物群，从生物标志物角度阐释“滋补肾阴”与“温补肾阳”延缓衰老作用机制的异同。研究工作分为三个部分：（1）源于自然衰老大鼠的血清代谢组学生物标志物研究：通过对2月~20月龄，即从成年大鼠到老年大鼠自然衰老过程的代谢轮廓谱分析，从内源性代谢物整体轮廓角度表征与大鼠增龄相关的生物标志物，发现与衰老密切相关磷酸酯酶A调控的磷脂胆碱类物质在大鼠自然衰老过程中显著降低，有望成为表征衰老状态的生物标志物；（2）更年期综合症和糖脂代谢病等与增龄相关疾病的临床血清代谢组学研究：来源于氨基乙酰-tRNA生物合成，甘氨酸、丝氨酸和苏氨酸代谢，三羧酸循环，缬氨酸、亮氨酸和异亮氨酸生物合成和苯丙氨酸代谢途径的内源性代谢物与更年期综合征患者年龄具有一定相关性；与线粒体损伤密切相关的酰基肉毒碱可表征糖脂代谢病患者的衰老程度，且与患者肾脏功能密切相关。（3）对补肾的经典方剂六味地黄丸和金匮肾气丸在传统中医中延缓衰老的用药历史及历代本草表述进行系统分析，采用高分辨质谱明确了他们的化学物质基础，通过系统药理学平台预测了六味地黄丸和金匮肾气丸中主要化合物的ADME参数，并通过网络药理学方法构建了六味地黄丸和金匮肾气丸的“药物-分子-靶点-通路”调控网络，比较它们延缓衰老作用机制的异同。