平性活血药调节寒、热不同血瘀证的物质基础及分子机制研究

It is very important for the improvement and development of the theory of Chinese herb to research the nature of the Chinese herb with the neutral nature. We have proved that the Chinese herb with the neutral nature have characteristics of Two-way apply and conditioned dominance. Recently there are two scientific problems need to resolve, one is “what are the material foundations of Chinese herb with neutral nature to regulate the cold and hot syndromes? ”, the other is “How Chinese herb with neutral nature regulate the cold and hot syndromes from gene level?”. In order to resolve the two scientific problems; we will select 3 Chinese herbs with the function of improving blood circulation and obvious neutral nature as research object, use HPLC-MS/MS to test the decoction, and serum of rats of normal control group, normal administration group, cold syndrome control group, cold syndrome administration group, hot syndrome control group and hot syndrome administration group, according chromatographic information and mass spectrometry, analyze the serum fingerprint of Chinese herbs in different syndromes; we use gene chip to test and analyze the difference of genome-wide expression profiles of rat of normal control group, cold syndrome model control group, cold syndrome administration group, hot syndrome model control group and hot syndrome administration group, use the Real-time PCR and Western Blot to prove the results of gene chip, find the molecular mechanisms of Chinese herbs with the function of improving blood circulation and neutral nature to regulating the cold and hot syndromes of blood stasis, finally reveal the scientific connotation of the characteristics of Two-way apply and conditioned dominance of Chinese herb with the neutral nature.

开展平性药药性研究，对于中药药性理论的完善和发展具有重要意义。项目组前期证实了平性药具有“双向适用、条件显性”药性特征。目前，对于“平性药调节寒、热不同病证的物质基础是什么？”，“平性药如何从基因水平调节寒、热不同病证?”这两个科学问题，还没有很好的解决。本项目围绕以上科学问题，以“平性”药性特征显著的3味平性活血药为研究对象；采用HPLC-MS/MS技术，检测水煎液，正常对照组、正常给药组、寒证对照组、寒证给药组、热证对照组、热证给药组血清，根据色谱信息和质谱裂解规律，对不同血瘀证下中药血清图谱进行分析，揭示平性活血药调节寒、热不同血瘀证物质基础及共性规律；采用基因芯片技术，检测和分析正常对照组、寒证模型组、寒证给药组、热证模型组、热证给药组大鼠全基因组表达谱的差异，揭示平性活血药调节寒、热不同血瘀证大鼠的分子机制；最终阐明平性药“双向适用、条件显性”的科学内涵。

本项目围绕 “平性药调节寒、热不同病证的物质基础是什么？”，“平性药如何从基因水平调节寒、热不同病证?”这两个科学问题，以“平性”药性特征显著的3味平性活血药为研究对象；采用HPLC-MS/MS技术，检测水煎液，正常对照组、正常给药组、寒证对照组、寒证给药组、热证对照组、热证给药组血清，根据色谱信息和质谱裂解规律，对不同血瘀证下中药血清图谱进行分析，揭示平性活血药调节寒、热不同血瘀证物质基础及共性规律；采用基因芯片技术，检测和分析正常对照组、寒证模型组、寒证给药组、热证模型组、热证给药组大鼠全基因组表达谱的差异，揭示平性活血药调节寒、热不同血瘀证大鼠的分子机制；最终阐明平性药“双向适用、条件显性”的科学内涵。项目研究结果对蒲黄、王不留行、桃仁3味平性活血药体外化学成分进行了表征，对蒲黄、王不留行、桃仁3味平性活血药在寒热不同证型大鼠体内入血成分及代谢产物进行了表征，鉴定了部分入血成分，在一定程度上明确了3味平性药调节寒、热不同血瘀证的物质基础及共有规律。明确了王不留行主要通过血小板活化、局灶性粘连、Rap1信号传导途径等多条途径调节寒凝血瘀证；通过RNA转运、内吞、氧化磷酸化等多条途径调节瘀热互结证。蒲黄组主要通过血小板活化、Rap1信号传导途径、血管平滑肌收缩等多条途径调节寒凝血瘀证；通过甘油磷脂代谢、醚脂质代谢、RNA降解等多条途径调节瘀热互结证。桃仁组主要通过花生四烯酸代谢、细胞因子与细胞因子受体的相互作用、血清素能突触等多条途径调节寒凝血瘀证；通过近端小管碳酸氢盐回收、甘油磷脂代谢、色氨酸代谢等多条途径调节瘀热互结证。在一定程度上阐明了平性药“双向适用，条件显性”的科学内涵。对于完善平性药性理论，指导平性药临床应用具有一定的科学意义。同时，本项目主要针对具有活血作用的平性药进行研究，所建立的研究模式，对于其他功效的平性药药性研究具有很好的示范作用，可进一步推广应用于其他平性药及寒、热药药性科学内涵研究。