基于热生成网络节点探索中药寒热属性的分子基础和评测体系

This subject is a continuing study for the last terminated fund from the National Natural Science Foundation of China. However, the objective biological activity of cold or hot property attributed to a particular herb still remains to be evaluated and validated, and thus has become one of the key scientific problems of traditional Chinese medicine to be resolved. While implementing the grant "correlation between cold and hot attributes of Chinese herbs and TRP channel" from the National Natural Science Foundation of China, our team found that activation/adjustment to the Thermo-TRP channel could be one of the most important ways for Chinese herbs to express their property and characterization. Given brown adipose tissue (BAT) is the terminal biological effector of a variety of energy metabolism regulation factor (including Thermo-TRP) in the body, this project is put forward for the first time, based on the expression changes of the key molecules of pgc-1 alpha and UCP1 on the BAT thermogenesis control core pathways, under basal metabolism or the load condition, to uncover the cold/hot attributes of traditional Chinese herbs associated with heat-generated molecular basis. In addition, we plan to adopt hypersensitive thermocouple synchronous detection analysis technology, with the heat-generated key point of BAT as well as the core body and surface temperatures as parameters, to undermine the corresponding relationship between cold/hot attributes of traditional Chinese herbs and the thermogenesis and heat dissipation, with the collected signals from the three specific sites, under different experimental conditions, to try to establish a set of quantifiable trinity of multidimensional evaluation standard system, and thus lay a solid foundation for full exploration of modern scientific connotation of cold/hot attributes of Chinese herbs.

本项目是对前一个结题的国家自然科学基金的延展性研究。我们课题组在实施前一个国家自然科学科学基金中发现：激活/调节TRP寒热感受通道（Thermo-TRP）是中药寒热药性表征的重要方式之一；棕色脂肪组织（BAT）热生成是包括Thermo-TRP在内的机体多种能量代谢调节因子的生物效应终端环节。本项目继续从能量代谢变化规律入手，深入探索机体能量代谢体系中TRP通路后（Post-TRP）产热的关键网络节点-PGC-1α和UCP1与中药寒热药性的生物学关联机制，揭示中药寒热属性与热生成相关的分子基础。同时基于超敏热偶同步检测分析技术，以BAT热生成关键点位为核心，结合机体的中心及体表部位，通过基础和负荷实验条件下中药作用后的动物上述三个部位特定位点热学信号的捕获，挖掘中药寒热属性与产热和散热环节的对应关系及相互间的规律性联系，尝试建立一套客观表征中药寒热属性的可量化的三位一体的多维评测的标准体系。

本课题主要研究的是中医药理论中的中药寒热药性问题，基于能量代谢环节对寒热中药药性进行初步研究和评价，同时基于整体和离体实验揭示其可能的分子机制。研究结果显示，在恒温环境下，给小鼠灌胃吴茱萸水煎液、肉桂水煎液等，或皮下注射高良姜素等，小鼠尾温升高，而肛温和背温下降，此时小鼠散热增加，一段时间后，随着能量代谢的增加，小鼠肛温、背温开始升高，产热增加。而小鼠的尾温在给药后，有明显的升高，在短时间内下降。在给予热性药肉桂的主要成分桂皮醛后，小鼠的肛温、背温和尾温均持续升高。给小鼠灌胃黄连水煎液、黄芩水煎液等，或皮下注射热性中药的成分后，小鼠的肛温、背温和尾温在升高后开始下降，并持久维持在较低的温度。在给于寒性药黄柏水煎液后，小鼠的肛温、背温和尾温均迅速增加，在120min后温度开始下降，并保持在较低的温度。灌胃热性药的实验组中，小鼠BAT中的PGC-1α和UCP1的mRNA表达量明显增加；白芷组、肉桂组、吴茱萸组、三七组等的UCP1mRNA的表达量显著上调。而灌胃寒性药的实验组中，PGC-1α和UCP1mRNA的表达量显著下降。部分灌胃热性药的小鼠BAT中UCP1蛋白表达量上升；部分灌胃寒性药的小鼠BAT进行UCP1蛋白表达量下降显著。离体试验中，发现部分寒性中药和中药成分下调UCP1和PGC-1α的表达；而部分热性药和活性成分上调UCP1和PGC-1α基因和蛋白表达。