基于血清药物化学-肠道菌群的蒙药香青兰调血脂药效物质及作用机制研究

Cardiac and cerebrovascular disease is one of the main fatal diseases at home and abroad. Its morbidity, disability rate and case fatality are increasing year by year. The abnormal blood lipids is closely related to the occurrence and development of cardio-cerebrovascular disease, especially coronary heart disease. It is one of the components of metabolic syndrome. Mongolian medicine Dracocephali Moldavicae Herba is commonly used in the prevention and treatment of coronary heart disease, hypertension, atherosclerosis and other diseases. Compared with the chemical drugs that regulate blood lipids, it has the advantages of having a long history of application, rich resources, accurate efficacy, and small side effects. However, there are few in-depth research reports on Dracocephali Moldavicae Herba. The pharmacodynamics substance basis and mechanism is not yet clear. This project will focuse on combining traditional isolation, identification and activity screening with modern and advanced research methods such as serum pharmacochemistry and intestinal flora. It will utilize animal models and cell models of abnormal blood lipids as biological systems. Using techniques such as LC-MS/MS, multidimensional chromatography and gene sequencing, we will separate and identify active ingredients in vitro, moving components in serum and detect intestinal flora. It will clarify the material basis and mechanism of Mongolian medicine Dracocephali Moldavicae Herba and will provide a reference for pharmacodynamics substance basis and mechanism of ethnic medicine.

心脑血管疾病是目前国内外主要致死性疾病之一，其发病率、致残率以及病死率呈逐年上升趋势。血脂异常与心脑血管病，尤其是冠心病的发生和发展密切相关，是代谢综合征的组成成分之一。蒙药香青兰民间常用于治疗冠心病、高血压、动脉粥样硬化等疾病，与调血脂的化药相比较，具有应用历史悠久、资源丰富、疗效确切、副作用小的优势。但是目前对香青兰的较为深入的研究报道较少，药效物质基础和作用机制尚不明确。本项目重点将传统的分离鉴定和活性筛选与血清药物化学及肠道菌群研究等现代先进的研究方法相结合，借助血脂异常动物模型和细胞模型为生物体系，运用液质联用、多维色谱、基因测序等技术，分离与鉴定体外活性成分、血清中移行成分阐明蒙药香青兰调血脂的药效物质，并通过研究肠道菌群多样性，说明调血脂可能的作用机制，为民族药的药效物质基础及作用机制研究提供参考。

蒙药香青兰在调血脂方面具有显著的作用，但其药效物质及作用机制尚未明确，严重制约了该药的临床应用、深入开发以及标准的建立。. 本项目利用体外活性筛选、体外化学成分鉴定、网络药理学、代谢组学和肠道菌群的方法，对蒙药香青兰的药效物质和作用机制进行阐释。. 通过活性筛选得到蒙药香青兰乙酸乙酯部位（EAD）能够降低TC和TG的含量，抑制HepG2细胞的脂质堆积；通过液质联用在香青兰乙酸乙酯部位初步鉴定出46个化学成分；网络药理学结果表明槲皮素、山奈酚、木犀草素、芹菜素、咖啡酸、丁香酸、柚皮素、齐墩果酸、山奈酚-3-O-半乳糖苷和对羟基肉桂酸可能是EAD调血脂的潜在活性化合物，EAD调血脂常涉及胰岛素抵抗、PPAR信号通路、脂肪细胞脂肪分解的调节、甘油酯代谢、精氨酸和脯氨酸代谢等通路；体内实验结果显示EAD对高脂血症大鼠血液中TC、TG、LDL-C含量具有显著降低作用，同时升高HDL-C的含量，还可逆转由高脂肪饲料引起的AST、ALT和LEP的增加，以及ADPN的下降趋势，对肝组织病理切片显示，EAD可以缓解高脂血症大鼠的肝脂肪变性；通过血清代谢组学研究发现血清样品中差异代谢物主要为L-丙氨酸、肌酸、肌酐、L-酪氨酸、L-脯氨酸、胆酸盐、甘胆酸盐等，主要影响初级胆汁酸生物合成、精氨酸和脯氨酸代谢、色氨酸代谢、泛酸和辅酶A生物合成等通路；通过盲肠内容物16s高通量测序发现嗜胆菌属、萨特氏菌属、乳酸杆菌属、梭状芽胞杆菌属XIVa、瘤胃球菌属、绿脓杆菌属、粪杆菌属、链球菌属、帕拉普氏菌属等菌属是EAD直接影响高脂模型大鼠菌群差异的关键菌；肠道微生物群与血清代谢标志物之间的相关性分析发现EAD干预后关联程度较大的差异菌群和差异代谢物分别是布劳特氏菌属、瘤胃球菌属、乳酸杆菌属、L-酪氨酸、丙氨酸、生物素等。. 通过以上研究阐明了蒙药香青兰调血脂活性部位、药效物质及作用机制，为蒙药香青兰的临床应用、深入开发以及标准的建立奠定基础，也为民族药的药效物质基础及作用机制研究提供参考。