基于“五味合化”理论研究酸甘化阴法生津止泻效应的生物分子网络调控机制

The integration and transformation of the five flavors theory is characteristic theory of traditional Chinese medicine,sour and sweet transforming yin law is an important manifestation of the theory. Relevant pre-clinical and experimental studies confirmed that the method has better promoting production of body fluid and antidiarrheal efficacy for syndrome of qi and yin eficiency of children diarrhea, but its mechanism is not well understood. Biomolecular network analysis technology based on network biology and network pharmacology provides a new path for deeperly revealing sour and sweet transforming yin law mechanism of action, and is expected to achieve a breakthrough. Under the guidance of traditional Chinese medicine theory,the subject translates the interaction of syndrome and drug into the interaction of disease-related molecules and drug intervention molecules in network target.By the means of gene chip, bioinformatics, systems biology and network pharmacology, using the technology of virtual identification reverse molecular docking, fluorescence quantitative PCR, Western Blot and other ,the subject explore the intrinsic relationship between network targets and regulation of biomolecular networks of promoting production of body fluid and antidiarrheal efficacy of the sour and sweet transforming yin law (the tailoring of ginseng dark plum decoction).Though the experiment of molecules - cells - the overall level reverse verifying ,to reveal the molecular network regulation mechanism of promoting production of body fluid and antidiarrheal efficacy of sour and sweet transforming yin law,from the gene expression and water metabolism angles,which further illustrates the scientific connotation of sour and sweet transforming yin law,and provides richer scientific basis for its clinical use.

五味合化理论是中医学传统特色理论，酸甘化阴法是该理论的重要体现。相关前期临床和实验研究证实，酸甘化阴法治疗小儿腹泻气阴两虚证具有良好的生津止泻效应，但其机理研究还不够深入。基于网络生物学和网络药理学的生物分子网络分析技术为更深入揭示酸甘化阴法的作用机制提供了新的路径，有望取得突破性进展。本课题在中医理论指导下，将病证与药物的相互作用转化为网络靶标上疾病相关分子与药物干预分子的相互作用，通过基因芯片、生物信息学、计算系统生物学、网络药理学等手段，运用分子对接、荧光定量PCR、Western Blot等方法技术，探讨酸甘化阴法（人参乌梅汤加味）复方及其拆方治疗腹泻病的网络靶标及与AQP调控相关信号生物分子网络的内在关系，并通过分子-细胞-整体水平验证，从基因表达、水液代谢的角度揭示酸甘化阴法生津止泻效应的分子网络调控机制，以进一步阐明酸甘化阴法的科学内涵，并为其临床运用提供更丰富的科学依据

酸甘化阴法是中医五味合化理论的重要体现，网络生物学与网络药理学技术为深入揭示酸甘化阴法的作用机制提供了新的路径。本课题借助腹泻动物模型，采用基因芯片、分子对接、RT-PCR、HPLC-DAD指纹图谱等技术和方法，探讨人参乌梅汤加味复方及其拆方治疗腹泻病的网络靶标、有效成分及与AQP调控相关信号分子生物网络的整合调节机制，以阐明其生津止泻效应的分子作用机制。结果：①人参乌梅汤加味具有良好的生津止泻效应，其中酸甘化阴的配伍发挥主要作用。②人参乌梅汤加味可通过调节 AQP4 及相关调控因子 Na + /K + -ATPase，PKC 基因的表达水平达到生津止泻的作用效应。③全基因芯片研究显示人参乌梅汤加味可调控腹泻模型大鼠结肠差异基因的表达，筛选基因涉及离子转运、免疫防御、细胞代谢、信号转导等多种腹泻相关功能及信号通路。④人参乌梅汤加味的主要有效物质在于“酸味药+甘味药”，主要有效部位在石油醚萃取部位，主要药效物质基础是人参皂苷类和有机酸类成分。⑤计算机虚拟筛选展示了人参乌梅汤加味27种活性成分与12种相关靶蛋白相互作用分子机制。研究初步表明，人参乌梅汤加味通过复方多成分、多靶点、多途径调控差异基因、水通道蛋白相关信号表达而发挥其生津止泻效应。项目从药物-靶标-疾病水平，初步揭示了酸甘化阴法（人参乌梅汤加味）生津止泻效应的分子网络调控机制，为进一步诠释中医五味合化理论的现代科学内涵奠定了基础。