基于脑-肠互动机制探索百合治疗百合病的科学内涵

Depression is associated with Lily Disease. The Classic decoctions with main component of lily in an ancient Chinese medical book < Jin Gui Yao Lue > written by Zhang Zhongjing are quite effective for Lily Disease treatment. However, the underlying mechanism is uncertain. Gastrointestinal dysfunction is one of common symptoms of Lily Disease. Modern research has discovered the existence of "brain-gut" axis, which explained why the depression patients usually accompany with irritable bowel syndrome (IBS). Preliminary research showed that Lily improved depression combined with IBS. Therefore we postulate that dysfunction of brain-gut crosstalk is the key factor for the morbidity of lily disease, and lily treat Lily Disease by keeping brain-gut homeostasis. In this research, we will testify the crosstalk of "brain-gut" based on the classic rat model of depression accompanied with IBS, by which the effect of lily on the brain-gut hormones（Corticotrophin Releasing Factor (CRF), 5-hydroxytryptamine (5-HT)） and the underlying mechanism of regulation of brain-gut crosstalk will be investigated, especial on the response and expression of CRF and its receptor, using the methods and techniques of pharmacology, genomics and bioinformatics. Subsequently, gene chip technology will be used to screen the genetic markers of depression patients accompanied with IBS whom diagnosed with Yin deficiency both in heart and lung in Chinese medicine-, and the outcomes shall be compared with the that from animal experiments to verify the specific bio-marks of Lily disease. This research aims to elucidate the mechanism of lily treatment on lily disease, interpret the relationships between lily disease and depression accompanied with IBS and throw lights on the clinical diagnostics and therapeutics of Lily Disease .

抑郁症与中医百合关联密切，张仲景《金匮要略》以百合为主药的百合类经方治疗百合病有效，其科学内涵尚待阐明。胃肠不适是百合病常见病症之一，现代研究证实了"脑-肠"轴的存在，揭示了临床抑郁症多伴肠易激综合征（IBS）的原因。我们已证实了百合具有抗抑郁兼改善IBS的作用，由此设想脑-肠互动功能失衡是百合病发病的基础，百合调节脑-肠互动功能应是其治疗百合病的机制之一。我们将通过抑郁和IBS模型验证脑-肠互动；复制并完善抑郁伴IBS模型，运用现代方法和技术，研究百合等药物对脑-肠激素（促肾上腺皮质激素释放因子（CRF）、5-羟色胺（5-HT）等）的影响，重点在CRF及受体的反应和表达上，探讨其调节脑-肠互动功能的作用机制；进一步与临床抑郁症伴IBS（中医辨证心肺阴虚型）患者的基因标志物进行对比分析和验证。本研究旨在诠释百合病的科学本质，揭示百合病与抑郁症伴IBS的联系，为临床辩证诊治百合病提供依据。

目的：基于脑肠互动机制，探索百合总皂苷(LS)、知母提取物(TI)和百合知母汤提取物(LT)治疗百合病的科学内涵。方法：建造并评价抑郁伴肠应激综合征的脑肠共病动物模型；进行LS、TI、LT的提取、含量检测以及质量标准和量效关系研究。通过分析药效机制，探讨LS、TI、LT对参与脑肠轴功能的HPA轴、单胺类神经递质、脑肠肽功能、中枢谷氨酸功能、炎症反应的改善效果。分析实验大鼠大脑以及血液全基因表达谱，进行GO和Pathway分析并验证。结果：抑郁伴IBS共病模型表现出抑郁症与IBS共病状态，且具有良好的重现性和特异性。LS平均含量不得低于5.91mg/g。TI总皂苷含量为44.58%，总黄酮含量为7.26%。LT按含百合皂苷1、百合皂苷2、知母皂苷BⅡ不得少于1.18％、0.82％和23.53％。量效LS为100mg/kg，TI为220mg/kg,LT为239.4mg/kg。 LS、TI、LT具有明显改善模型抑郁样行为以及IBS症状，改善模型大鼠海马和结肠病理改变，调节中枢与结肠单胺类神经递质功能，抑制HPA轴功能过度激活。降低模型大鼠谷氨酸毒性以及调节脑肠肽功能和肠道炎症反应的功效。全基因表达谱分析显示LS、TI、LT调节多个与抑郁症以及炎症发生相关的细胞信号传导通路。验证结果显示，LS、TI、LT上调模型大鼠海马以及结肠PPI3K-AKT-NF-κB-BCL-2 信号通路，最终下调caspase 3 mRNA表达抑制细胞凋亡以及炎症过程，下调长时程抑制信号通路的基因表达，改善海马学习记忆过程，下调多巴胺突触通路以及上调5-HT突触通路基因表达，上调单胺类神经递质含量，同时加速突触囊泡转运递质能力，改善突触转运能力。结论:综上所述，LS、TI、LT通过调节中枢与结肠PPI3K- AKT-NF-κB-BCL-2-caspase 3信号通路以及的分子机制，抑制细胞凋亡，抑制炎症发生。调节GNAQ、PLCB4、COMT、TPH1、SLC18A1、SNAP25、SLC7A1基因表达改善学习记忆过程和中枢单胺类神经递质含量，改善中枢神经突触前体与突触后体囊泡运输递质能力，加快神经递质信号的传递过程。通过上述的分子机制作用，调节HPA轴功能，中枢和结肠5-HT功能以及脑肠肽功能系统，同时抑制中枢神经毒性和肠道炎症的效果。最终达到改善抑郁以及IBS共病（百合病）状态的药效。