基于网络药理学与脑脊液谱效学相结合研究天泰1号多靶协同抗AD的物质基础

Alzheimer's disease (AD) is a typical complex multi-factorial disease. The effect of single-target drug was unsatisfactory. TCM treatment strategy of multi-target therapeutics has guiding significance for the treatment of AD. Tiantai No.1 is a Hospital preparation which was developed on the basis of AD "gan xu sui kui" new pathological hypotheses and long-term clinical experience. Pharmacological studies have shown that Tiantai No.1 intervenes AD pathology process by promoting its core pathological product degradation, anti- endoplasmic reticulum stress, anti-oxidation, anti-apoptosis, enhancing cholinergic activity and hippocampal synaptic plasticity, and so on. Tiantai No.1 has a significant curative effect on AD, but the material basis of its efficacy has not been clear.Therefore, network pharmacology will be used to predict active ingredients and targets of Tiantai No.1 against Alzheimer disease, molecular docking technology and network topology feature analysis will be used to clarify the network relationship of "ingredient - target - disease". Cerebrospinal fluid spectrum-efficiency study,as a new method,will be used to study the relationship between the “quantity” of chemical composition in cerebro-spinal and “effect” in cell experiment. Clearify the active ingredients of Tiantai No.1 against Alzheimer disease by integrating network pharmacology and Cerebrospinal fluid spectrum-efficiency study,and systematically elucidate the scientific compatibility among components.Screening the active ingredients for different target through the network characteristics to achieve the new forms multi-component Chinese Medicine.

阿尔茨海默病（AD）是一种典型多因素复杂疾病，针对单一靶点研发的临床一线药物疗效不尽人意。多靶协同的中医策略对AD防治具有指导性意义。天泰1号是基于AD“肝虚髓亏”中医病理新假说和长期临床经验研制的医院制剂。研究显示其通过促进核心病理产物降解、拮抗内质网应激、抗氧化、抗凋亡、增强胆碱能活性和突触可塑性等多种途径干预AD。天泰1号临床效果显著，但其药效物质基础尚未清楚。鉴此，本项目采用网络药理学筛选天泰1号抗AD活性成分群、靶点群，通过分子对接、网络拓扑分析明确“活性成分-靶点-疾病”的网络关系；以体内直接作用物质为切入点，以脑脊液谱效学新方法，研究脑脊液药物成分“量”的动态变化与细胞模型“效”的关系。整合网络药理学和脑脊液谱效学，明确天泰1号抗AD的物质基础；通过网络拓扑分析，解析天泰1号多组分协同抗AD的整合机制，为实现由传统中药组方向新型中药复方（组分中药）转化提供科学依据。

阿尔茨海默病（AD）是一种典型多因素复杂疾病，多靶协同的中医策略对AD防治具有指导性意义。医院制剂天泰1号临床效果显著，但其药效物质基础尚未清楚。鉴此，本项目采用网络药理学与脑脊液谱效学相结合的方法探讨天泰1号抗AD的活性成分和靶点。网络药理学研究共获得87个抗AD潜在活性成分（主要包括人参皂苷、黄连生物碱、吴茱萸生物碱、二苯乙烯苷类及姜黄素类成分等）和68个潜在靶点（主要包括雌激素受体、前列腺素合成酶、乙酰胆碱酯酶、有丝分裂原激活蛋白激酶、糖原合成酶激酶、过氧化物酶体增殖激活受体、乙酰胆碱受体、肿瘤坏死因子等），涉及44条主要信号通路。天泰1号脑脊液谱效学研究共鉴定了53个成分，其中脑脊液中26个原型移行成分；药效学研究表明，与模型组相比，天泰1号含药脑脊液能显著提高模型细胞的存活率，降低细胞上清液中IL-1β、IL-6、TNF-α、MDA的含量，增加SOD、GSH-Px的活性和SYP的浓度（P<0.05）；“谱-效”回归分析结果表明，人参皂苷Rb1与细胞增殖，小檗碱、去甲氧基姜黄素与SOD活性，人参皂苷Rb1、巴马亭与GSH-Px活性，人参皂苷Rg1与突触素生成均成正相关；巴马亭、药根碱、人参皂苷Rd与MDA释放，柠檬苦素与TNF-α释放，去甲氧基姜黄素、巴马亭与IL-1β释放，巴马亭、黄连碱、小檗碱、柠檬苦素与IL-6释放均成负相关。活性验证表明，巴马亭、人参皂苷Rd、人参皂苷Rb1分别降低MAPK14、CASP3、PTGS2 mRNA表达，小檗碱降低NOS2和AChE mRNA表达，去甲氧基姜黄素、药根碱、黄连碱、吴茱萸碱分别升高GSK3B、ESR1、PPARG、CHRM1 mRNA表达。整合网络药理学和脑脊液谱效学研究结果，我们推测天泰1号抗AD主要活性成分为人参皂苷、黄连生物碱、吴茱萸生物碱、姜黄素类成分，作用靶点包括AR、ESR1、PTGS2、NOS2、ACHE、MAPK14、GSK3B、PPARG等。天泰1号抗AD活性成分和靶点的明确，为医院制剂的二次开发提供科学依据。