基于辛润理论和水通道蛋白的蚕沙“和胃化湿”机理研究

Faeces Bombyx is pungent in flavor. At present, its effects of “regulating stomach and resolving dampness” as well as “moisturizing dryness” could not be fully explicated by the theory that “pungent taste having dispersing effect” and it comes from the feces. Accordingly, it was less used in clinic and even recommended to be abandoned. This project is based on the theory of TCM and preliminary experiments. We put forward the hypothesis that: As it’s pungent, Faeces Bombyx can adjust the aquaporins to disperse and scattered moisture, and then adjust the intestinal flora and substance metabolism in brain and urine, play the role of “regulating stomach and resolving dampness”, In order to achieve the purpose of “moistening”. By using the technology of Q RT-PCR, Western Blot, PCR-DGGE and MD, the effect on aquaporins, intestinal flora and metabonomics will be investigated to clarify the mechanism of “regulating stomach and resolving dampness”; The physical basis of “regulating stomach and resolving dampness” will be explored by PCA, PLS-DA and so on. And then the change rule of the composition of Faeces Bombyx in its forming process will be investigated. By using gene detection technology the main bacterial species in Faeces Bombyx will be tested to clarify the scientific, rational and feasibility of feces as Chinese Medicine. This project is to research the effect of Faeces Bombyx Based on “Xinrun Theory” and Aquaporins in order to extend the theory “pungent taste having dispersing effect”. It will provide a new perspective to explain the effect of other pungent drugs. It will also provides an example for the study of other fecal drugs, and promote the inheritance and development of traditional Chinese Medicine coming from feces.

蚕沙味“辛”，现代仅以辛散（行）理论并不能全面阐述其“和胃化湿”机制及“润燥”效果，同时由于其来源于粪便，故在临床上应用较少，甚至建议废除。本项目在中医理论及前期实验基础上，提出“蚕沙味‘辛’而调节水通道蛋白，布散水湿，进而调节肠道菌群及脑部、尿液中物质代谢，发挥“和胃化湿”的作用，以达到‘润’的目的”的假说，采用qRT-PCR、Western Blot、PCR-DGGE、MD技术，考察蚕沙对水通道、肠道菌群、脑神经递质及尿液代谢产物的影响，阐明“和胃化湿”机制；采用PCA、PLS-DA等探索“和胃化湿”的物质基础，考察蚕沙在形成过程中成分的变化规律，利用基因测序分析蚕沙中主要细菌种类，阐明粪便蚕沙作为中药的科学性、合理性、可行性。本项目基于辛润理论和水通道研究蚕沙作用机理，扩展了辛散（行）理论，为辛味中药功效阐释提供新的视角；也为其他粪便类中药研究提供示范，助推粪便类中药的传承与发展。

本项目在中医理论及前期实验基础上，提出“蚕沙味‘辛’而调节水通道蛋白，布散水湿，进而调节肠道菌群及脑部、尿液中物质代谢，发挥“和胃化湿”的作用，以达到‘润’的目的”的假说，考察蚕沙对水通道、肠道菌群、内源性代谢产物的影响，阐明“和胃化湿”机制。结果表明，蚕沙一方面通过调节水通道蛋白AQP的表达发挥“化湿”作用，而这种作用与蚕沙介导“cAMP-PKA-CREB”信号通路有关；另一方面，蚕沙可通过平衡肠道菌群、安定神志等发挥“和胃”作用；再一方面，蚕沙整体上通过干预内源性物质的“代谢”发挥“化湿和胃”作用。在“桑叶——蚕沙”过程中，味“辛”的挥发性成分的“质”和“量”均发生明显改变，同时活性成分异槲皮苷等的含量也显著增加；微生物分析结果表明，蚕沙中的优势菌门为变形菌门、放线菌门；在属水平上，蚕沙主要含有葡萄球菌属、勒克菌属、泛菌属等。本项目基于辛润理论和水通道研究蚕沙作用机理，扩展了辛散（行）理论，为辛味中药功效阐释提供新的视角；也为其他粪便类中药研究提供示范，助推粪便类中药的传承与发展。