基于肾病综合征模型及AVP-NOS/NO-AQP2途径研究当归芍药散活血利水作用机制及物质基础

当归芍药散是活血利水治法的代表方剂，通过前期对痛经、肝硬化腹水等疾病模型的研究，表明其活血利水作用与精氨酸加压素途径有关。但活血作用和利水作用二者如何关联及相关的物质基础等仍然不清楚。本项目以当归芍药散临床疗效确切的肾病综合征疾病模型为研究切入点，提出当归芍药散治疗肾病综合征的主要作用环节是AVP-NOS/NO-AQP2系统的工作假说，以整体动物和细胞实验研究当归芍药散及其不同配伍的干预作用，阐明当归芍药散的主要靶点和作用机制；以含药血清指纹图谱和重点药效指标构建“谱-效”关系，确定待敲除目标成分；采用成分敲除策略，以不同时间段组分为单位逐一敲除目标成分，再由体外实验考察目标成分的活性，以此辨识主要有效成分。本课题研究为进一步深入、全面地阐明当归芍药散活血利水作用机制和物质基础奠定基础，并为其他中药复方的研究提供思路和方法借鉴。

As Danggui Shaoyao San (DSS) is a classical formula, which representatives the therapeutic role of invigorating blood and promoting diuresis. Our preliminary studies on dysmenorrhea and cirrhosis indicated that the mechanism of DSS treating on blood stasis and water stagnation was by influencing arginine vasopressin (AVP) system. But the link between the effect of invigorating blood and the effect of promoting diuresis is remains unknown, the pharmacodynamic material basis is unclear. Our project carry out on DSS and nephrotic syndrome as the pointcut to clarify the hypothesis: the mechanism of DSS treating for nephrotic syndrome may be by influencing AVP-NOS/NO-AQP2 system. To clarify the targets and mechanism of DSS, we investigate the certain indicators of DSS and its different compatibility group by whole animal and cell experiments. We proposed the tactic of “multi-dimensional spectrum-effect relationship combined with component knockout”to identify the main pharmacodynamic material basis through the following steps: first, constructing the “spectrum-effect” relationship between fingerprint of serum containing DSS and the key efficacy indicators to determine the target components; second, by the developed strategy based on knockout of bioactive components using a combination of column chromatography and preparative HPLC for knocking out the target components; third, investigating the activity of target components in vitro. This research lay the foundation on the study of mechanism and pharmacodynamic material basis of DSS and provides idea and method for reference.