基于肠道微生态和代谢组学的丹参清心除烦功效物质基础与生物学机制研究

The traditional efficacy "Qīngxīn chú fán" of Salvia miltiorrhiza has been applied in ancient times, such as in the treatment of "insomnia", "epilepsy". Modern physicians also take this efficacy on the treatment of insomnia and depression. However, its functional material basis and mechanism need further to be explained. This project intends to take animal models of insomnia and depression for example to study the functional substance basis and mechanism of Salvia miltiorrhiza's "Qīngxīn chú fán". Research works in preparation are as follows: the first is to evaluate effect of Salvia miltiorrhiza on insomnia and depression. The second is to construct fingerprints of gut flora of insomnia and depression model animals via gut microbiology research technology in further to study the regulatory effect of Salvia miltiorrhiza on gut microecology. The third is to study the intervention effects of Salvia miltiorrhiza on gut brain-related metabolic profile, potential endogenous biomarkers and metabolic pathway via metabolomics, speculating the possible pathways. The forth is to analyze the correlation among intestinal flora, metabolites and biochemical indicators, exploring the pathogenesis of the model and the action mechanism of Salvia miltiorrhiza. The fifth is to reveal metabolic processes and metabolites of Salvia miltiorrhiza in vivo and verify the action mechanism of Salvia miltiorrhiza in vitro.The ultimate aim is to reveal the characteristic advantage and scientific connotation of the traditional efficacy "Qīngxīn chú fán" of Salvia miltiorrhiza.

丹参清心除烦功效在古代已有应用历史，如用于“不寐”、“癫”等证，现代医家亦取对丹参该功效用于治疗失眠和抑郁，并取得良好疗效，但其功效物质基础与作用机制亟待深入研究。本课题拟以失眠和抑郁为研究切入点，开展丹参清心除烦功效物质基础和作用机制研究：①采用肠道微生态研究技术，构建失眠与抑郁模型动物的肠道菌群结构和指纹图谱，研究丹参对肠道微生态的调节作用；②运用代谢组学方法和技术研究丹参对失眠和抑郁模型动物体内肠脑相关代谢物谱干预效应、靶标调控、代谢通路影响，推测可能作用途径；③运用相关分析方法，对肠道菌群、肠脑代谢物、生化指标之间的相关性进行深入分析，探讨其发病机制与丹参的作用机制；④采用药物代谢动力学研究方法揭示丹参体内效应物质及其代谢转化过程，并用分子生物学研究方法对丹参的作用机制进行验证。以期较为系统地回答丹参清心除烦的整体调节作用与作用机制，阐明丹参清心除烦功效的特色优势与科学内涵。

一、项目的背景：.失眠具有较高的患病率，而寻求治疗失眠的有效药物尤为迫切，丹参清心除烦治疗失眠的疗效确切，亟待阐明其功效物质与作用机制。.二、主要研究内容及结果：.1、采用回流提取的方法制备得到丹参水提物和丹参醇提物，丹参水提物中总酚酸含量为16.74%；丹参80%乙醇提取物中总酚酸含量为11.62%，总丹参酮含量为3.79%。.2、通过建立PCPA致失眠大鼠模型，评价丹参对其睡眠的改善作用，研究表明：PCPA致失眠大鼠模型建立成功，丹参能改善其失眠状态；PCPA致失眠模型大鼠HPA轴相关激素（CRH、ACTH、CORT）、炎症因子（NF-κB、TNF-α、IL-1β、IL-6）、神经递质（ 5-HT、Glu、GABA、Ach ）水平出现异常，丹参可部分减弱其异常变化。PCPA致失眠模型大鼠体内出现代谢紊乱，丹参能改善其代谢紊乱。.3、通过建立盐酸林可霉素诱导的肠道菌群失调大鼠模型，探索肠道菌群以5-HT为媒介与大脑之间的联系，研究表明：盐酸林可霉素诱导的肠道菌群失调大鼠模型建立成功；肠道菌群失调模型大鼠脑组织（大脑皮质、脑干、海马）出现病理变化，大鼠出现失眠现象；肠道菌群失调模型大鼠体内HPA轴相关激素（CRH、ACTH、CORT）、炎症因子（NF-κB、TNF-α、IL-1β、IL-6）水平出现异常；肠道菌群失调可影响5-HT的合成及代谢（抑制大鼠大脑皮质及血清中5-HT的合成，促进大脑皮质、海马、血清、结肠中5-HT的代谢5-HT合成及代谢）。.4、通过建立PCPA致失眠大鼠模型，评价丹参对其肠道菌群的调节作用，研究表明：PCPA致失眠模型大鼠结肠出现病理变化，出现肠道菌群失调，丹参能改善结肠病理状态及肠道菌群失调；丹参可促进失眠模型大鼠大脑皮质、海马及血清中5-HT合成，抑制5-HT代谢；促进脑干中5-HT合成及代谢。.三、科学意义：.揭示失眠症的肠道微生态及肠脑相关类代谢物的变化；.阐明丹参通过改善肠道微生态与神经内分泌途径抗失眠的5-HT机制及可能物质基础.四、发表论文4篇，其中SCI论文3篇，中文核心论文1篇。