基于“脑-肠-菌轴”的酸枣仁多组分养心安神协同作用机制研究

Insomnia is attracting more and more attention due to its high incidence. Ziziphi Spinosae Semen (ZSS) is commonly used in traditional Chinese medicine for the treatment of insomnia with character of “Nourishing the Heart and Calming Mind”. Especially, it is estimated to exert this function by the combination of multiple components. However, most of them are difficult to penetrate the blood-brain barrier, and the action mechanism has not been properly elucidated. Modern researches show that intestinal microflora can influence the occurrence and development of insomnia by interacting with "brain-gut axis". According to the above foundings, it is proposed that: The function of ZSS for nourishing heart and calming mind is closely related with the synergistic effects of the multiple components contained in ZSS on regulation of "brain-gut-microbiota axis". The following studies will be carried out: (1) To reveal the intervention of different components in ZSS on the combination of intestinal microflora in the insomnia-like rats based on the gene sequencing technology; (2) To clarify the effects of various components in ZSS on the intestinal barrier and immune function and its mechanism by analyzing the intestinal permeability and immune cytokines; (3) To reveal the effects and mechanisms of multiple components in ZSS on central nervous system through the expressions of neurotransmitters, hormones and cytokines in the brain; (4) Based on the multivariate statistical method, the synergistic mechanism of the ZSS for treating insomnia will be comprehensively clarified. The ultimate goal of this study is to elucidate the biological mechanism of ZSS for “Nourishing the Heart and Calming Mind” systematically from regulation of "brain-gut-microbiota axis", which will provide scientific basis for the clinical use and drug development of insomnia.

失眠症因其发病率日渐增高而备受关注。酸枣仁是中医临床治疗失眠症的常用药味，养心安神功效确切，且含有的多元物质呈现出独特的效应特点，但大多难以穿透血脑屏障，作用机制尚未得到合理阐明。现代研究显示，肠道菌群可通过与“脑-肠轴”交互作用影响失眠症的发生发展。据此提出：酸枣仁养心安神功效的发挥与其多类型化学组分协同调节“脑-肠-菌轴”功能密切相关。拟开展研究工作：（1）基于基因测序技术，揭示酸枣仁各组分对失眠模型动物肠道菌群结构的干预作用；（2）通过对肠道通透性、免疫因子等的分析，揭示酸枣仁各组分对肠道屏障、免疫功能的影响及其作用机制；（3）通过脑内相关递质、激素、细胞因子的分析，揭示酸枣仁各组分对中枢神经系统的调控效用与机制；（4）基于多变量统计方法，整合分析酸枣仁多组分协同作用机制。以期从“脑-肠-菌轴”角度较为系统地揭示酸枣仁养心安神功效发挥的生物学机制，为临床用药及新药创制提供科学依据。

酸枣仁为中医临床治疗失眠症的常用药味，养心安神功效确切，但其所含的活性组分大多难以穿透血脑屏障，作用机制尚未得到合理阐明。现代研究显示，肠道菌群可通过与“脑-肠轴”交互作用影响失眠症的发生发展。据此，本项目基于“脑-肠-菌轴”角度，分别从肠道菌群结构、肠道屏障/免疫功能、脑内相关因子和蛋白表达等方面，关联分析“脑-肠-菌轴”与失眠症发生发展的内在联系及酸枣仁的干预作用。.研究结果显示，酸枣仁可改善失眠引起的大鼠脑及肠组织病理改变，回调神经营养因子、炎症因子水平，改善失眠大鼠的学习记忆能力，回调失眠模型大鼠肠道棒状杆菌属_1（Corynebacterium_1）、红蝽菌科_UCG-002（Coriobacteriaceae_UCG-002）、unclassified_f__Erysipelotrichaceae、norank_f__Erysipelotrichaceae和Allobaculum相对丰度变化，干预失眠大鼠粪便及盲肠内容物短链脂肪酸含量降低趋势。进一步采用“伪无菌鼠”模型证实肠道菌群在酸枣仁防治失眠过程中发挥重要作用。.血浆及尿液进行代谢组学分析结果显示，酸枣仁可不同程度地调节失眠模型大鼠18种代谢差异物趋向正常水平，其主要参与牛磺酸和次牛磺酸代谢、D-谷氨酰胺和D-谷氨酸代谢、苯丙氨酸代谢、戊糖和葡萄糖醛酸相互转化、花生四烯酸代谢等途径。失眠相关差异菌群和生物标志物共变异分析结果显示，酸枣仁显著回调的5个肠道菌属与代谢途径主要为氨基酸代谢的多种代谢差异物具有显著相关性，提示酸枣仁可通过调整代谢谱和肠道菌群组成来改善失眠症状。.以酸枣仁中含有的代表性活性组分酸枣仁皂苷A和斯皮诺素为研究对象，比较正常大鼠和失眠模型大鼠对酸枣仁活性组分体内过程的影响，以及不同活性组分配伍前后的体内过程变化。结果显示：失眠模型大鼠因肠屏障功能受损，酸枣仁皂苷A、斯皮诺素相对生物利用度较正常大鼠显著提高，且其整体达峰时间较快；酸枣仁脂肪油组分可促进酸枣仁皂苷A及斯皮诺素的吸收。.项目研究成果在国内外学术刊物上发表论文9篇，其中SCI源期刊收录4篇，申请国家发明专利1项；研究成果获中国自然资源学会优秀科技奖；培养研究生2名。项目负责人郭盛入选江苏省“333”高层次人才培养工程。